U.S. patent application number 11/328483 was filed with the patent office on 2009-05-14 for compositions and methods for detection of antibody binding to cells.
This patent application is currently assigned to The Trustees of the University of Pennsylvania. Invention is credited to Donald L. Siegel.
Application Number | 20090123475 11/328483 |
Document ID | / |
Family ID | 27487687 |
Filed Date | 2009-05-14 |
United States Patent
Application |
20090123475 |
Kind Code |
A9 |
Siegel; Donald L. |
May 14, 2009 |
Compositions and methods for detection of antibody binding to
cells
Abstract
The invention includes Rh(D) binding proteins, including
antibodies, and DNA encoding such proteins. Methods of generating
such proteins and DNAs are also included.
Inventors: |
Siegel; Donald L.;
(Lansdale, PA) |
Correspondence
Address: |
DRINKER BIDDLE & REATH;ATTN: INTELLECTUAL PROPERTY GROUP
ONE LOGAN SQUARE
18TH AND CHERRY STREETS
PHILADELPHIA
PA
19103-6996
US
|
Assignee: |
The Trustees of the University of
Pennsylvania
|
Prior
Publication: |
|
Document Identifier |
Publication Date |
|
US 20060177440 A1 |
August 10, 2006 |
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Family ID: |
27487687 |
Appl. No.: |
11/328483 |
Filed: |
January 9, 2006 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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11064174 |
Feb 22, 2005 |
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11328483 |
Jan 9, 2006 |
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09848798 |
May 4, 2001 |
6858719 |
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11064174 |
Feb 22, 2005 |
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09240274 |
Jan 29, 1999 |
6255455 |
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09848798 |
May 4, 2001 |
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08884045 |
Jun 27, 1997 |
5876925 |
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09240274 |
Jan 29, 1999 |
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60028550 |
Oct 11, 1996 |
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60081380 |
Apr 10, 1998 |
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Current U.S.
Class: |
424/143.1 ;
435/320.1; 435/334; 435/69.1; 530/388.22; 536/23.53 |
Current CPC
Class: |
C07K 2317/55 20130101;
C07K 16/005 20130101; C07K 16/34 20130101; C12N 15/1013 20130101;
C12N 15/1037 20130101; C07K 16/08 20130101; C40B 40/02
20130101 |
Class at
Publication: |
424/143.1 ;
435/069.1; 435/320.1; 435/334; 530/388.22; 536/023.53 |
International
Class: |
A61K 39/395 20060101
A61K039/395; C07H 21/04 20060101 C07H021/04; C12P 21/06 20060101
C12P021/06; C12N 5/06 20060101 C12N005/06; C07K 16/28 20060101
C07K016/28 |
Goverment Interests
GOVERNMENT SUPPORT
[0002] This invention was supported in part by a grant from the
U.S. Government (NIH Grant No. P50-HL54516) and the U.S. Government
may therefore have certain rights in the invention.
Claims
1-21. (canceled)
22. A method of detecting the presence of an antigen-bearing moiety
on a cell comprising providing a mixture comprising a population of
cells and a population of bacteriophage expressing a known first
antibody on the surface of said bacteriophage, wherein the presence
of said antigen-bearing moiety on said cells is indicated by
binding of said first antibody to at least two of said cells
causing said bacteriophage to also bind to said at least two of
said cells, wherein when a second antibody is added to said mixture
which is specific for said bacteriophage said second antibody binds
to bacteriophage bound to said at least two of said cells causing
the cells to agglutinate, said agglutination being an indication of
the presence of said antigen-bearing moiety on said cell, which
antigen-bearing moiety is specific for said first antibody.
23. A method of identifying an antigen-bearing moiety on a cell
comprising providing a mixture comprising a population of cells and
a population of bacteriophage expressing a known first antibody on
the surface of said bacteriophage, wherein the presence of said
antigen-bearing moiety on said cells is indicated by binding of
said first antibody to at least two of said cells causing said
bacteriophage to also bind to said at least two of said cells,
wherein when a second antibody is added to said mixture which is
specific for said bacteriophage said second antibody binds to
bacteriophage bound to said at least two of said cells causing said
cells to agglutinate, wherein said agglutination identifies said
antigen-bearing moiety as being an antigen-bearing moiety specific
for said first antibody.
24. A method of detecting the presence of an antigen-bearing moiety
on a cell comprising providing a mixture comprising a population of
cells and a population of bacteriophage expressing a known first
antibody on the surface of said bacteriophage, wherein the presence
of said antigen-bearing moiety on said cell is indicated by binding
of said first antibody to at least two of said cells causing said
bacteriophage to also bind to said at least two of said cells,
adding said mixture to a microtube containing inert particles and a
second antibody specific for said bacteriophage, allowing said
mixture to sediment under the force of gravity, and observing the
location of cell in said microtube, wherein strong agglutination of
the cells is indicated by the cells being located upon or within a
top layer of said inert particles which strong agglutination is an
indication of the presence of said antigen-bearing moiety on said
cell, which antigen-bearing moiety is specific for said first
antibody.
25. A method of identifying an antigen-bearing moiety on a cell
comprising providing a mixture comprising a population of cells and
a population of bacteriophage expressing a known first antibody on
the surface of said bacteriophage, wherein the presence of said
antigen-bearing moiety on said cell is indicated by binding of said
first antibody to at least two of said cells causing said
bacteriophage to also bind to said at least two of said cells,
adding said mixture to a microtube containing inert particles and a
second antibody specific for said bacteriophage, allowing said
mixture to sediment under the force of gravity, and observing the
location of cells in said microtube, wherein strong agglutination
of cells is indicated by the cells being located upon or within a
top layer of said inert particles which strong agglutination
identifies said antigen-bearing moiety as being an antigen-bearing
moiety specific for said first antibody.
26. A method of detecting the presence of an antigen-bearing moiety
on a cell comprising providing a mixture comprising a population of
cells and a population of bacteriophage expressing a known first
antibody on the surface of said bacteriophage, wherein the presence
of said antigen-bearing moiety on said cell is indicated by binding
of said first antibody to at least two of said cells causing said
bacteriophage to also bind to said at least two of said cells,
adding said mixture to a microtube containing inert particles which
have bound thereto a second antibody specific for said
bacteriophage, allowing said mixture to sediment under force of
gravity, wherein captured cells are located upon or within a top
layer of said inert particles, the presence of said captured cells
being an indication of the presence of an antigen-bearing moiety on
said cell, which antigen-bearing moiety is specific for said first
antibody.
27. A method of identifying an antigen-bearing moiety on a cell
comprising providing a mixture comprising a population of cells and
a population of bacteriophage expressing a known first antibody on
the surface of said bacteriophage, wherein the presence of said
antigen-bearing moiety on said cell is indicated by binding of said
first antibody to at least two of said cells causing said
bacteriophage to also bind to said at least two of said cells,
adding said mixture to a microtube containing inert particles which
have bound thereto a second antibody specific for said
bacteriophage, allowing said mixture to sediment under force of
gravity, wherein captured cells are located upon or within a top
layer of said inert particles, the presence of said captured cells
identifying said antigen-bearing moiety on said cell as being
specific for said first antibody.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is entitled to priority pursuant to 35
U.S.C. .sctn.119(e) to U.S. provisional patent application
60/081,380, which was filed on Apr. 10, 1998, and is a
continuation-in-part of U.S. application Ser. No. 08/884,045, filed
Jun. 27, 1997, which application is entitled to priority under 35
U.S.C. .sctn.119(e) to U.S. Provisional Application No. 60/028,550,
filed on Oct. 11, 1996.
FIELD OF THE INVENTION
[0003] The field of the invention is generation of binding
proteins.
BACKGROUND OF THE INVENTION
[0004] The ability to produce monoclonal antibodies has
revolutionized diagnostic and therapeutic medicine. Monoclonal
antibodies are typically produced by immortalization of
antibody-producing mouse lymphocytes thus ensuring an endless
supply of cells which produce mouse antibodies. However, for many
human applications, it is desirable to produce human antibodies.
For example, it is preferable that antibodies which are
administered to humans for either diagnostic or therapeutic
purposes are human antibodies since administration of human
antibodies to a human circumvents potential immune reactions to the
administered antibody, which reactions may negate the purpose for
which the antibody was administered.
[0005] In addition, there exists certain situations where, for
diagnostic purposes, it is essential that human antibodies be used
because other animals are unable to make antibodies against the
antigen to be detected in the diagnostic method. For example, in
order to determine the Rh phenotype of human red blood cells
(RBCs), human sera that contains anti-Rh antibody must be used
since other animals cannot make an antibody capable of detecting
the human Rh antigen.
[0006] The production of human antibodies in vitro by immortalizing
human B lymphocytes using Epstein Barr virus (EBV)-mediated
transformation or cell fusion has been fraught with technical
difficulties due to the relatively low efficiency of both
EBV-induced transformation and cell fusion when compared with the
murine system. To overcome these problems, processes have been
developed for the production of human antibodies using M13
bacteriophage display (Burton et al., 1994, Adv. Immunol.
57:191-280). Essentially, a cDNA library is generated from "iRNA
obtained from a population of antibody-producing cells. The mRNA
encodes rearranged immunoglobulin (Ig) genes and thus, the cDNA
encodes the same. Amplified cDNA is cloned into M13 expression
vectors creating a library of phage which express human Fab
fragments on their surface. Phage which display the antibody of
interest are selected by antigen binding and are propagated in
bacteria to produce soluble human Fab Ig. Thus, in contrast to
conventional monoclonal antibody synthesis, this procedure
immortalizes DNA encoding human Ig rather than cells which express
human Ig.
[0007] There are several difficulties associated with the
generation of antibodies using bacteriophage. For example, many
proteins cannot be purified in a non-denatured state, in that
purification procedures necessarily involve solubilization of
protein which may render some proteins permanently denatured with
concomitant destruction of antigenic sites present thereon. Such
proteins thus cannot be bound to a solid phase and therefore cannot
be used to pan for phage bearing antibodies which bind to them. An
example of such a protein is the human Rh antigen.
[0008] To solve the problem, a method was developed wherein intact
RBCs were used as the panning antigen (Siegel et al., 1994, Blood
83:2334-2344). However, it was discovered that since phage are
inherently "sticky" and RBCs express a multitude of antigens on the
cell surface, a sufficient amount of phage which do not express the
appropriate antibody on the surface also adhere to the RBCs, thus
rendering the method impractical for isolation of phage which
express antibody of desired specificity.
[0009] De Knif et al. (1995, Proc. Natl. Acad. Sci. USA
92:3938-3942) disclose a method of isolating phage encoding
antibodies, wherein antibody-expressing phage are incubated with a
mixture of antigen-expressing cells and cells which do not express
antigen. The antibody-expressing phage bind to the
antigen-expressing cells. Following binding with phage, a
fluorescently labeled antibody is added specifically to the
antigen-expressing cells, which cells are removed from the mixture
having antibody-expressing phage bound thereto. The isolation of
fluorescently labeled cells is accomplished using the technique of
fluorescently-activated cell sorting (FACS), an expensive and
time-consuming procedure.
[0010] There remains a need for a method of isolating recombinant
proteins, preferably antibodies, which is rapid and economical, and
which will provide a vast array of protein-binding proteins useful
for diagnostic and therapeutic applications in humans.
SUMMARY OF THE INVENTION
[0011] The invention relates to an isolated protein having an amino
acid sequence comprising a sequence selected from the group
consisting of SEQ ID NOs: 1-69 and 139-181. In one embodiment, the
isolated protein is an antigen-binding protein. In one aspect, the
antigen is human Rh(D) protein. In another embodiment, the binding
protein has an amino acid sequence selected from the group
consisting of SEQ ID NOs: 1-69 and 139-181. In one aspect, the
binding protein is an antibody. In another aspect, the said
antibody comprises a heavy chain having an amino acid sequence
selected from the group consisting of SEQ ID NOs: 1-28 and 139-153.
In still another aspect, the antibody comprises a light chain
having an amino acid sequence selected from the group consisting of
SEQ ID NOs: 29-69 and 154-181. In yet another aspect, the antibody
comprises a heavy chain having an amino acid sequence selected from
the group consisting of SEQ ID NOs: 1-28 and 139-153 and a light
chain having an amino acid sequence selected from the group
consisting of SEQ ID NOs: 29-69 and 154-181.
[0012] In another embodiment of the isolated binding protein, the
binding protein is an antibody fusion protein.
[0013] In another embodiment of the isolated protein, the protein
is substantially purified.
[0014] The invention also includes an isolated DNA encoding the
isolated protein of the invention. In one embodiment, the isolated
DNA has a nucleotide sequence selected from the group consisting of
SEQ ID NOs: 70-138 and 182-224. In another embodiment, the DNA is
substantially purified.
[0015] The invention also includes an isolated DNA encoding a
protein obtained by generating a synthetic DNA library in a virus
vector expressing said protein; adding a magnetic label to cells
expressing said antigen-bearing moiety; incubating virus expressing
said protein with said magnetically labeled cells in the presence
of an excess of non-labeled cells which do not express said
antigen-bearing moiety to form a mixture, wherein said virus binds
to said magnetically labeled cells; isolating virus bound cells
from said mixture and obtaining DNA encoding said protein
therefrom. In one embodiment, the DNA has a nucleotide sequence
selected from the group consisting of SEQ ID NOs: 70-138 and
182-224.
[0016] The invention further includes a substantially pure protein
obtained by generating a synthetic DNA library in a virus vector
expressing said protein; adding a magnetic label to cells
expressing said antigen-bearing moiety; incubating virus expressing
said protein with said magnetically labeled cells in the presence
of an excess of non-labeled cells which do not express said
antigen-bearing moiety to form a mixture, wherein said virus binds
to said magnetically labeled cells; isolating virus bound cells
from said mixture and isolating said protein therefrom. In one
embodiment, the protein has an amino acid sequence selected from
the group consisting of SEQ ID NOs: 1-69 and 139-181.
[0017] The invention also includes a substantially pure preparation
of a protein obtained by expressing said protein from DNA encoding
said protein, wherein said DNA is obtained by generating a
synthetic DNA library in a virus vector expressing said protein;
adding a magnetic label to cells expressing said antigen-bearing
moiety; incubating virus expressing said protein with said
magnetically labeled cells in the presence of an excess of
non-labeled cells which do not express said antigen-bearing moiety
to form a mixture, wherein said virus binds to said magnetically
labeled cells; isolating virus bound cells from said mixture and
obtaining DNA encoding said protein therefrom. In one embodiment,
the protein has an amino acid sequence selected from the group
consisting of SEQ ID NOs: 1-69 and 139-181.
[0018] The invention further relates to a method of isolating a DNA
encoding a multi-subunit protein which binds to an antigen-bearing
moiety. This method comprises [0019] (a) generating a phage display
library comprising a plurality of virus vectors. A first of the
virus vectors comprises a first heterologous DNA encoding a subunit
of the protein and expresses the subunit on the surface thereof. A
second of the virus vectors comprises a second heterologous DNA
encoding a different subunit of the protein and expresses the
different subunit on the surface thereof. [0020] (b) adding a
magnetic label to cells bearing the antigen-bearing moiety on their
surface. [0021] (c) incubating the phage display library with the
magnetically labeled cells in the presence of an excess of
non-labeled cells which do not express the antigen-bearing moiety
to form a mixture. The first and second virus vectors thereby bind
to the magnetically labeled cells. [0022] (d) isolating
magnetically labeled cells from the mixture. The first and second
virus vectors are thereby isolated from the mixture. [0023] (e)
obtaining the first heterologous DNA from the first virus vector.
[0024] (f) ligating at least the portion of the first heterologous
DNA encoding the subunit and at least the portion of the second
heterologous DNA encoding the different subunit to form a hybrid
heterologous DNA. [0025] (g) generating a hybrid virus vector
comprising the hybrid heterologous DNA and expressing the subunit
and the different subunit of the protein on the surface thereof.
[0026] (h) adding a magnetic label to cells bearing the
antigen-bearing moiety on their surface. [0027] (i) incubating the
hybrid virus vector with the magnetically labeled cells in the
presence of an excess of non-labeled cells which do not express the
antigen-bearing moiety to form a mixture. The hybrid virus vector
thereby binds to the magnetically labeled cells. [0028] (j)
isolating magnetically labeled cells from the mixture. The hybrid
virus vector is thereby isolated from the mixture. [0029] (k)
obtaining DNA encoding the protein from the isolated virus vector.
The DNA is thereby isolated.
[0030] The invention also relates to a method of isolating a
multi-subunit protein which binds to an antigen-bearing moiety.
This method comprises [0031] (a) generating a phage display library
comprising a plurality of virus vectors. A first of the virus
vectors comprises a first heterologous DNA encoding a subunit of
the protein and expresses the subunit on the surface thereof. A
second of the virus vectors comprises a second heterologous DNA
encoding a different subunit of the protein and expresses the
different subunit on the surface thereof. [0032] (b) adding a
magnetic label to cells bearing the antigen-bearing moiety on their
surface. [0033] (c) incubating the phage display library with the
magnetically labeled cells in the presence of an excess of
non-labeled cells which do not express the antigen-bearing moiety
to form a mixture. The first and second virus vectors thereby bind
to the magnetically labeled cells. [0034] (d) isolating
magnetically labeled cells from the mixture. The first and second
virus vectors are thereby isolated from the mixture. [0035] (e)
obtaning the first heterologous DNA from the first virus vector.
[0036] (f) ligating at least the portion of the first heterologous
DNA encoding the subunit and at least the portion of the second
heterologous DNA encoding the different subunit to form a hybrid
heterologous DNA. [0037] (g) generating a hybrid virus vector
comprising the hybrid heterologous DNA and expressing the subunit
and the different subunit of the protein on the surface thereof.
[0038] (h) adding a magnetic label to cells bearing the
antigen-bearing moiety on their surface. [0039] (i) incubating the
hybrid virus vector with the magnetically labeled cells in the
presence of an excess of non-labeled cells which do not express the
antigen-bearing moiety to form a mixture. The hybrid virus vector
thereby binds to the magnetically labeled cells. [0040] (j)
isolating magnetically labeled cells from the mixture. The hybrid
virus vector is thereby isolated from the mixture. [0041] (k)
isolating the protein from the isolated virus vector. The protein
is isolated.
BRIEF DESCRIPTION OF THE DRAWINGS
[0042] FIG. 1 is a diagram of a strategy for cell-surface Fab-phage
panning using magnetically-activated cell sorting.
[0043] FIG. 2 is a graph depicting cell-surface biotinylation of
human RBCs.
[0044] FIG. 3 is a series of graphs which validate the
antigen-positive, antigen-negative cell separation procedure of the
invention.
[0045] FIG. 4 is an image of a microplate agglutination assay
wherein anti-Rh(D) Fab/phage agglutination titer was measured.
[0046] FIG. 5 is an image of a microplate agglutination assay
showing determination of Rh(D) binding epitope for selected
anti-Rh(D) Fab/phage clones.
[0047] FIG. 6 is an image depicting the use of Fab/phage antibodies
in a gel card assay.
[0048] FIG. 7 comprises FIGS. 7A and 7B. FIG. 7A is a dendrogram
which depicts the relationship among the anti-Rh(D) heavy chains
described herein in Example 2. The 28 unique heavy chain clones are
organized by V.sub.H family, V.sub.H germline gene, and VDJ
rearrangement. Each heavy chain clone is identified by a numeral
preceded by a letter ("B" through "E") which denotes its germline
gene. The 28 heavy chains comprised 12 distinct VDJ regions,
designated VDJ1 - VDJ12. Clones with identical VDJ joins putatively
result from intra-clonal diversity of 12 original B lymphocytes.
FIG. 7B is an alignment of the CDR3 regions of the anti-Rh(D) heavy
chains.
[0049] FIG. 8 comprises FIGS. 8A, 8B, and 8C. FIG. 8A is an
alignment of anti-Rh(D) heavy chains to their nearest germline V,
D, and J genes. Also illustrated are the putative intermediate
heavy chain sequences (Ca, Cb, Da, Db, Dc). The number of
nucleotide differences from a germline V.sub.H is tabulated to the
right of each sequence. In general, D segments showed poor homology
with known D genes so mutations were not scored in these regions.
Replacement mutations are indicated with letters, silent mutations
are indicated as "*", identities are indicated as ".", and
insertions are indicated as "-". Sequences derived from the 5'
V.sub.H primers used in library construction are indicated as
">". FIG. 8B is an alignment of the four VH3 genes utilized by
anti-Rh(D) heavy chains. FIG. 8C is a dendrogram which depicts the
relationship among human VH3 family germline genes, and illustrate
relatedness of VH3-21, VH3-30. VH3-33, and VH3-30.3 and the
surprising restriction in V.sub.H gene usage. The VH3-30.5 gene is
present in only certain haplotypes and is identical to VH3-30.
[0050] FIG. 9 is an ontogenic tree of anti-Rh(D) heavy chains
constructed using nucleotide alignment data. Circles represent
isolated and sequenced clones, and diamonds represent putative
intermediates. The number of nucleotide mutations from its germline
V.sub.H gene is indicated in parentheses below the clone name. The
distance along the horizontal axis represents the degree of
mutation (including J segments) within the constraints of the
diagram.
[0051] FIG. 10 comprises FIGS. 10A and 10B. FIG. 10A is an
alignment of anti-Rh(D) .kappa. light chains to their nearest
germline V and J genes, and indicates predominance of DPK-9 usage
from the V.sub..kappa.I family. Nomenclature for clones is similar
to that for heavy chains but uses the letters "F" through "I". FIG.
10B is an alignment of the four V.sub..kappa. genes utilized by
anti-Rh(D) light chains. Symbols are the same as those used in FIG.
8A.
[0052] FIG. 11 comprises FIG. 11A and 11B. FIG. 11A is an alignment
of anti-Rh(D) .lamda. light chains to their nearest germline V and
J genes. FIG. 11B is an alignment of the 10 V.sub..lamda. germline
genes utilized, and illustrates the use of a diverse set of
variable region genes derived from multiple families. However, all
of the clones use the identical J.sub..lamda. gene segment.
Nomenclature for the clones is similar to that for heavy chains but
uses the letters "J" through "S". Symbols are the same as those
used in FIG. 8A.
[0053] FIG. 12, comprising FIGS. 12A, 12B, and 12C, is a trio of
graphs which depict comparisons of variable region gene family
usage for anti-Rh(D)-specific clones and randomly-picked,
non-Rh(D)-binding clones from original .gamma..sub.1.kappa. and
.gamma..sub.1.lamda. non-selected libraries. Lightly-hatched bars
reveal heterogeneity in V.sub.H (FIG. 12A), V.sub..kappa. (FIG.
12B), and V.sub..lamda. (FIG. 12C) family representation before
selection for anti-Rh(D) specificity. Numbers above bars represent
absolute number of clones in that group.
[0054] FIG. 13 depicts the results of determinations of the Rh(D)
binding epitope of anti-Rh(D) Fab/phage clones. The five different
agglutination patterns obtained from screening all of the 53
Fab/phage clones are illustrated. The particular clones shown in
FIG. 13 are identified by their unique heavy chain/light chain
pairings using the nomenclature defined in FIGS. 7, 10, and 11. For
E1/M3, reactivity with additional Rh(D) variant cells is required
to distinguish its specificity for epD3 from that for epD9.
Inclusion of the category IVb cell permits the identification of a
new epitope designated "epDX".
[0055] FIG. 14 is matrix illustrating the genetic composition and
epitope specificity of anti-Rh(D) antibodies. The horizontal axis
represents the unique .gamma..sub.1 heavy chains and the vertical
axis represents the unique .lamda. and .kappa. light chains (based
on amino acid sequence). A shaded pattern at the intersection of a
heavy chain/light chain pair indicates the Rh(D) epitope
specificity observed for that Fab/phage antibody. A few clones gave
mixed patterns of reactivity as described herein. Although heavy
chains D1, D15, D16, and D17 differ in nucleotide sequence, these
chains have an identical amino acid sequence and thus comprise a
single column. Similarly, heavy chains C5 and C8 and A light chains
K1 and K2 encode the same proteins. The pairings of these 28 heavy
and 41 light chain nucleotide gene segments, which produced 53
unique Fab transcripts, encoded 43 different Fab proteins, as
indicated in the matrix.
[0056] FIG. 15, comprising FIGS. 15A, 15B, and 15C, depicts the
results of inhibition studies performed using recombinant
anti-Rh(D) antibodies. The figures show results of representative
experiments demonstrating the mutual inhibition of antibodies
directed at two different Rh(D) epitopes (in this example, epD3 and
epD6/7, FIGS. 15A and 15C), but not between an Rh(D) antibody and
an unrelated recombinant anti-RBC antibody (an anti-blood group B
antibody, FIG. 15B). In FIG. 15A, Rh(D)-positive RBCs were
incubated with soluble Fabs only, phage-displayed Fabs only, or
combinations of the two, as indicated. In FIG. 15B, Rh(D)-positive
RBCs that were blood group B were used. After washing, RBCs were
resuspended in anti-M13 antibody and assessed for agglutination
induced by phage-displayed Fabs. Soluble Fabs were used
"full-strength" while Fab/phage preparations were present in
limiting amounts to increase the sensitivity of the inhibition
assay, as described herein. In FIG. 15C, mutual inhibition of epD3
and epD6/7 anti-Rh(D) antibodies was demonstrated with
Rh(D)-positive RBCs, .gamma..sub.1.kappa. and .gamma..sub.1.lamda.
soluble Fabs, and light chain isotype-specific antisera (see text
for details). In these examples, the anti-epD3 and anti-epD6/7
antibodies were clones E1/M3 and D5/I3, respectively. The
anti-blood group B antibody was isolated from an IgG phage display
library made from the splenic B cells of a blood group O donor.
[0057] FIG. 16, comprising FIGS. 16A, 16B, and 16C, depict models
for Rh(D) antigen/antibody binding. A conventional model (depicted
in FIG. 16A) and a model described herein (depicted in FIG. 16B)
for Rh(D) antigen/antibody binding predict different combining
sites and genetic relationships between antibodies. As depicted in
FIG. 16C, if antibodies directed at different Rh(D) epitopes are
clonally related, then the expressed repertoire will differ between
Rh(D)-negative and partial Rh(D) individuals.
DETAILED DESCRIPTION
[0058] According to the present invention, there is provided a
novel method of isolating DNA encoding a protein and the protein
encoded thereby, wherein the protein is preferably an antibody,
which protein is capable of specifically binding to an
antigen-bearing moiety.
[0059] As exemplified herein but not limited thereto, the method
comprises generating bacteriophage which encode human antibodies.
Specifically in the present invention, anti-Rh(D) RBC Fab/phage
antibodies encoded by an M13 filamentous phage library are
obtained. The library is generated from antibody-producing cells
obtained from a hyperimmunized donor by first-obtaining cDNA
derived from mRNA expressed in the antibody-producing cells. Ig
encoding fragments of the cDNA are obtained using the polymerase
chain reaction (PCR) and primers specific for such fragments of
DNA. Ig-specific DNA so obtained is cloned into a bacteriophage.
Bacteriophage encoding the Ig fragments are panned against a
mixture of antigen-positive, biotinylated RBC-target cells
pre-coated with streptavidin-conjugated magnetic microbeads and
excess non-labeled RBCs. Bacteriophage which express antibodies on
the phage surface, which antibodies are specific for the target
cell antigen, bind to the labeled cells. These phage are separated
from phage which are bound to non-labeled cells and from phage
which are not bound to the cells using a magnetic column. Phage so
separated encode and display antibody specific for antigens on the
target cells.
[0060] To generate a phage antibody library, a cDNA library is
first obtained from mRNA which is isolated from cells which express
the desired protein to be expressed on the phage surface, e.g., the
desired antibody. cDNA copies of the mRNA are produced using
reverse transcriptase. cDNA which specifies Ig fragments are
obtained by PCR and the resulting DNA is cloned into a suitable
bacteriophage vector to generate a bacteriophage DNA library
comprising DNA specifying Ig genes. The procedures for making a
bacteriophage library comprising heterologous DNA are well known in
the art and are described, for example, in Sambrook et al. (1989,
Molecular Cloning: A Laboratory Manual, Cold Spring Harbor,
N.Y.).
[0061] A bacteriophage library may also be obtained using cDNA
rather than PCR-amplified Ig encoding fragments of cDNA. Generation
of a cDNA library is useful for the isolation of proteins which are
not antibodies, such as ligands and the like.
[0062] Bacteriophage which encode the desired protein, e.g., an
antibody, may be engineered such that the protein is displayed on
the surface thereof in such a manner that it is available for
binding to its corresponding binding protein, e.g., the antigen
against which the antibody is directed. Thus, when bacteriophage
which express a specific antibody are incubated in the presence of
a cell which expresses the corresponding antigen, the bacteriophage
will bind to the cell. Bacteriophage which do not express the
antibody will not bind to the cell.
[0063] For panning of bacteriophage, ie., selection of phage which
express the desired antibody, cells which express the corresponding
antigen are labeled with a detectable label such as biotin.
Streptavidin-conjugated magnetic beads are then added to the cells.
The cells are mixed with an excess of non-labeled cells which do
not express the antigen. This cell mixture is then incubated with
the phage library, wherein phage which express the antibody bind to
cells expressing the antigen. The presence of the excess
non-labeled cells in the mixture serves as a means of removing
bacteriophage which do not express the antibody but which might
otherwise bind to antigen-expressing cells non-specifically. The
details of the experimental procedures for practicing the present
invention are provided herein in the experimental detail
section.
[0064] Antigen-expressing cells having antibody-expressing phage
bound thereto are magnetically removed from the mixture. One
example of magnetic removal involves pouring the mixture of
magnetic and non-magnetic cells into a column in the selective
presence or absence of a magnetic field surrounding the column.
Alternatively, magnetic cells may be separated from non-magnetic
cells in solution by simply holding a magnet against the side of a
test tube and attracting the cells to the inner wall and then
carefully removing the non-magnetic cells from the solution.
[0065] Thus, the method of the invention involves a procedure for
enriching a population of recombinant phage for those expressing
specific phage-displayed ligands derived from natural or synthetic
phage DNA libraries by simultaneously performing negative and
positive selection against a mixture of magnetically-labeled
receptor-positive particles (i e., cells) and non-labeled
receptor-negative particles.
[0066] The terms "bacteriophage" and "phage" are used
interchangeably herein and refer to viruses which infect bacteria
By the use of the terms "bacteriophage library" or "phage library"
as used herein, is meant a population of bacterial viruses
comprising heterologous DNA, i.e., DNA which is not naturally
encoded by the bacterial virus.
[0067] The term "virus vector" includes a virus into which
heterologous DNA has been inserted. The virus vector may be a
bacteriophage or may be a eukaryotic virus.
[0068] By the term "target cell" as used herein, is meant a cell
which expresses an antigen against which the desired antibody is
sought.
[0069] By the term "panning" or "panned" as used herein, is meant
the process of selecting phage which encode the desired
antibody.
[0070] By the term "Fab/phage" as used herein, is meant a phage
particle which expresses the Fab portion of an antibody.
[0071] By the term "scFv/phage" are used herein, is meant a phage
particle which expresses the Fv portion of an antibody as a single
chain.
[0072] By "excess non-labeled cells" is meant an amount of
non-labeled cells which exceeds the number of labeled cells.
Preferably, the ratio of labeled cells to non-labeled cells is
about 1:2. More preferably, the ratio of labeled cells to
non-labeled cells is greater than about 1:4. Even more preferably,
the ratio of labeled cells to non-labeled cells is greater than
about 1:10.
[0073] While the method of the invention as exemplified herein
describes the generation of phage which encode the Fab portion of
an antibody molecule, the method should not be construed to be
limited solely to the generation of phage encoding Fab antibodies.
Rather, phage which encode single chain antibodies (scFV/phage
antibody libraries) are also included in the invention. Fab
molecules comprise the entire Ig light chain, that is, they
comprise both the variable and constant region of the light chain,
but include only the variable region and first constant region
domain (CH1) of the heavy chain. Single chain antibody molecules
comprise a single chain of protein comprising the Ig Fv fragment.
An Ig Fv fragment includes only the variable regions of the heavy
and light chains of the antibody, having no constant region
contained therein. Phage libraries comprising scFV DNA may be
generated following the procedures described in Marks et al., 1991,
J. Mol. Biol. 222:581-597. Panning of phage so generated for the
isolation of a desired antibody is conducted as described herein
for phage libraries comprising Fab DNA.
[0074] The invention should also be construed to include synthetic
phage display libraries in which the heavy and light chain variable
regions may be synthesized such that they include nearly all
possible specificities. Therefore, antibody-displaying libraries
can be "natural" or "synthetic" (Barbas, 1995, Nature Medicine
1:837-839; de Kruif et al. 1995, J. Mol. Biol. 248:97-105).
Antibody-displaying libraries comprising "natural" antibodies are
generated as described in the experimental example section.
Antibody-displaying libraries comprising "synthetic" antibodies are
generated following the procedure described in Barbas (1995, supra)
and the references cited therein.
[0075] The method of the invention should be further construed to
include generation of phage display libraries comprising phage
other than M13 as exemplified herein. Other bacteriophage, such as
lambda phage, may also be useful in the method of the invention.
Lambda phage display libraries have been generated which display
peptides encoded by heterologous DNA on their surface (Sternberg et
al., 1995, Proc. Natl. Acad. Sci. USA 92:1609-1613). Moreover, it
is contemplated that the method of the invention may be extended to
include viruses other than bacteriophage, such as eukaryotic
viruses. In fact, eukaryotic viruses may be generated which encode
genes suitable for delivery to a mammal and which encode and
display an antibody capable of targeting a specific cell type or
tissue into which the gene is to be delivered. For example,
retroviral vectors have been generated which display functional
antibody fragments (Russell et al., 1993, Nucl. Acids Res.
21:1081-1085).
[0076] The red blood cell antibodies to which antibodies may be
generated include, but are not limited to, Rh antigens, including
Rh(D), Rh(C), Rh(c), Rh(E), Rh(e), and other non-Rh antigens,
including red blood cell antigens in the Kell, Duffy, Lutheran and
Kidd blood groups.
[0077] Thus, the method of the invention is not limited solely to
the isolation of DNA encoding anti-Rh(D) antibodies, but rather may
be used for the isolation of DNA encoding antibodies directed
against any RBC antigen or other cell antigen, such as, but not
limited to, tumor-specific antigen, bacterial antigens, and the
like. The method of the invention is also usefull for typing
platelets by generating phage antibodies specific for a number of
clinically important platelet antigens, notably,
p1.sup.A1/p1.sup.A2, Bak.sup.a/Bak.sup.b, Pen.sup.A/Pen.sup.B, and
the like.
[0078] The invention is further usefull for typing donor white
blood cells for HLA antigens for the purposes of matching donors
and recipients for potential transplant matching in the case of
both solid (for example, kidney, heart, liver, lung) and non-solid
(for example, bone marrow) organ or tissue transplanting.
[0079] To detect binding of phage expressing antibody directed
against one of these non-red blood cell antigens, the non-red blood
cells may be agglutinated or trapped following the procedures
described herein for agglutination or trapping of red blood cells.
Prior to agglutination or trapping, the cells may be rendered
"visible" by staining or other labeling technique in order that
agglutination or trapping is apparent to the naked eye or
scanner.
[0080] The method of the invention is most useful for the
generation of a protein which binds to an antigen-bearing moiety,
where the antigen-bearing moiety is not easily purified in soluble
form. Thus, the antigen-bearing moiety includes antigens which are
associated with other structures, usually membranes in the cell
such as cell membranes or cell organelle membranes.
[0081] In accordance with the present invention, the
antigen-bearing moiety may be a protein, a lipid, a carbohydrate or
a nucleic acid, or it may be a complex of at least two of a
protein, a lipid, a carbohydrate and a nucleic acid, it being
appreciated that many antigen-bearing moieties in cells are not
comprised of one of these components alone. Preferably, the
antigen-bearing moiety is a membrane bound protein, such as an
antigen or a receptor protein. However, when the antigen-bearing
moiety is a carbohydrate, it may be a carbohydrate expressed on a
glycolipid, for example, a P blood group antigen or other
antigen.
[0082] By the term "antigen-bearing moiety" as used herein, is
meant a molecule to which an antibody binds.
[0083] By the term "antigen-binding protein" as used herein, is
meant a polypeptide molecule, such a an antibody, a fragment
thereof or an antibody fusion protein, which is capable of
specifically binding to another molecule.
[0084] By the term "antibody fusion protein" as used herein, is
meant a polypeptide molecule having an amino acid sequence which
comprises the amino acid sequence of a portion of an
antigen-binding protein. The portion of the antigen-binding protein
may, for example, be an entire antibody or a fragment thereof.
[0085] The method of the invention is also useful for the
generation of autoimmune antibodies such as those involved in
autoimmune hemolytic anemia (AIHA) (Siegel et al., 1994, Structural
analysis of red cell autoantibodies, Garratty (ed) Immunobiology of
Transfusion Medicine, Dekker, New York, N.Y.). Autoimmune
antibodies that are directed against cell antigens which are cell
surface membrane associated or cell organelle membrane associated
may be isolated using the technology described herein. Autoimmune
diseases and their associated antigens to which antibodies may be
isolated include, but are not limited to the following: Myasthenia
gravis (acetylcholine receptor; neurons), chronic inflammatory
demyelinating polyneuropathy (myelin; neurons), autoimmune thyroid
disease (thyroid stimulating hormone receptor; thyroid cells),
primary biliary cirrhosis (mitochondrial autoantigens; liver
mitochondria), idiopathic thrombocytopenic purpura (platelet
membrane integrins; platelets), pemphigus vulgaris (epidermal
antigens; epidermis), and Goodpasture's syndrome (basement membrane
antigens; kidney or lung cells).
[0086] In fact, the method of the invention is useful for the
isolation of DNA clones encoding any antibody directed against-an
antigen expressed on a cell, which cell can be labeled with a
magnetic label and which cell can be obtained in sufficient
quantities in an non-labeled form so as to provide an excess of
non-labeled cells as required in the assay.
[0087] Further, the method of the invention is not limited to the
isolation of DNA encoding antibodies but rather may also be used
for the isolation of DNA encoding other peptides or proteins having
specificity for cell proteins, such as, for example, but not
limited to, ligands which bind cell receptor proteins, peptide
hormones, and the like.
[0088] The invention should also not be construed as being limited
to the use of biotin as the cell-labeling agent. Other labels may
be used provided their addition to a cell does not disturb the
structural integrity of any surface proteins expressed thereon and
provided such labels permit the addition of a paramagnetic
microbead or other magnetic substance thereto. Other such labels
include, but are not limited to, cell surface proteins or
carbohydrates which can be directly derivitized with magnetic beads
that possess activated amine, carboxyl, or thiol groups. In
addition, dyes such as fluorescein or rhodamine may also be
covalently attached to cells in a manner similar to biotin and
magnetic beads coated with anti-dye antibodies may be attached
thereto.
[0089] The invention also includes a screening method which may be
used to isolate a DNA encoding a multi-subunit protein which binds
to an antigen-bearing moiety or, alternately, to isolate the
multi-subunit protein itself. The multi-subunit protein may, for
example, be an antibody or another immunoglobulin. It is well known
that antibodies and other immunoglobulins comprise multiple
subunits, often designated heavy and light chains.
[0090] According to this screening method, a phage display library
is generated, either as described herein or using other generally
known or hereafter-developed methods. The library comprises a
plurality of virus vectors, including a first virus vector which
comprises a first heterologous DNA encoding a subunit of the
protein. The first virus vector expresses the subunit on its
surface, either by itself or in association with one or more other
subunits of the protein. The library also comprises a second virus
vector which comprises a second heterologous DNA encoding a
different subunit of the protein. The second virus vector expresses
the different subunit on its surface, either by itself or in
association with one or more other subunits of the protein. A
magnetic label is added to cells bearing the antigen-bearing moiety
on their surface, and the labeled cells are incubated with the
phage display library in the presence of an excess of non-labeled
cells which do not express the antigen-bearing moiety. The first
and second virus vectors bind to the magnetically labeled cells,
owing to interaction(s) between the antigen and the subunits of the
protein expressed on the surface of the vectors.
[0091] After incubating the phage display library with the mixture
of cells, magnetically labeled cells are isolated from the mixture.
First and second virus vectors bound to the magnetically labeled
cells are thereby also isolated from the mixture. The virus vectors
are separated from the magnetically labeled cells (e.g. by
culturing the cells in a manner in which the virus vectors are
produced in the culture supernatant), and heterologous DNA is
obtained from virus vectors that adhered to the magnetically
labeled cells. The DNA may optionally be purified at this stage.
DNA isolated from the virus vectors that adhered to the
magnetically labeled cells includes the first heterologous DNA and
the second heterologous DNA.
[0092] At least the portion of the first heterologous DNA encoding
the subunit is ligated to at least the portion of the second
heterologous DNA encoding the different subunit to form a hybrid
heterologous DNA. For this purpose, it is advantageous that the
virus vector be constructed in such a way that the portion of the
first heterologous DNA encoding the subunit, the portion of the
second heterologous DNA encoding the different subunit, or both,
are flanked or surrounded by defined restriction endonuclease
cleavage sites. In such constructs, the portion of the first
heterologous DNA encoding the subunit may be removed, for example,
by treating the first heterologous DNA with restriction
endonucleases which specifically cleave the specific sites. This
portion may then be ligated, for example either directly or after
ligating a linker DNA thereto, to all or a portion of the second
heterologous DNA to generate the hybrid heterologous DNA.
[0093] The hybrid heterologous DNA is then used to generate a
hybrid virus vector comprising the hybrid heterologous DNA. The
hybrid virus vector expresses the subunit and the different subunit
of the protein on its surface. For example, if the first
heterologous DNA encodes an antibody light chain and the second
heterologous DNA encodes an antibody heavy chain, then the hybrid
virus vector may express an antibody comprising equal numbers of
heavy and light chains on its surface.
[0094] The hybrid virus vector is then incubated with the mixture
of magnetically labeled cells having the antigen-bearing moiety on
their surface and non-magnetically labeled cells which do not have
the antigen-bearing moiety on their surface. Owing to interactions
between the antigen and the subunits of the protein expressed on
the surface of the hybrid virus vector, the hybrid virus vector
binds with the magnetically labeled cells, and may therefore be
isolated from the mixture of cells by isolating magnetically
labeled cells from the mixture.
[0095] As described herein, hybrid virus vector particles are
isolated from the magnetically labeled cells. The isolated hybrid
virus vectors may be used as a source for obtaining either the
multi-subunit protein or the hybrid heterologous DNA (which encodes
the subunits of the protein), using standard methods.
[0096] The invention includes proteins and DNA encoding the same
which are generated using the methods described herein. To isolate
DNA encoding an antibody, for example, DNA is extracted from
antibody expressing phage obtained according to the methods of the
invention. Such extraction techniques are well known in the art and
are described, for example, in Sambrook et al. (supra).
[0097] The invention includes a number of isolated or substantially
purified, DNAs encoding antigen-binding proteins, such as
Rh(D)-binding proteins. For example, a DNA having a nucleotide
sequence comprising at least one of SEQ ID NOs: 70-138 and 182-224,
as described herein, is included. The isolated or substantially
purified nucleic acid may have a nucleotide sequence selected from
the group consisting of SEQ ID NOs: 70-138 and 182-224.
[0098] An "isolated DNA", as used herein, refers to a DNA sequence,
segment, or fragment which has been purified from the sequences
which flank it in a naturally occurring state, e.g., a DNA fragment
which has been removed from the sequences which are normally
adjacent to the fragment, e.g., the sequences adjacent to the
fragment in a genome in which it naturally occurs. The term also
applies to DNA which has been substantially purified from other
components which naturally accompany the DNA, e.g., RNA or DNA or
proteins which naturally accompany it in the cell.
[0099] The invention also includes a number of isolated or
substantially purified proteins, such as Rh(D)-binding proteins.
For example, a protein having an amino acid sequence comprising at
least one of SEQ ID NOs: 169 and 139-181, as described herein, is
included. The isolated or substantially purified protein may have
an amino acid sequence selected from the group consisting of SEQ ID
NOs: 1-69 and 139-181. The protein may be an antigen-binding
protein, such as an antibody which comprises a heavy chain having
an amino acid sequence selected from the group consisting of SEQ ID
NOs: 1-28 and 139-153, a light chain having an amino acid sequence
selected from the group consisting of SEQ ID NOs 29-69 and 154-181,
or both. The protein may also be, for example, an antibody fusion
protein.
[0100] An "isolated protein" as used herein, means a protein or
polypeptide which has been separated from components which
naturally accompany it in a cell. Typically, a protein or
polypeptide is isolated when at least 10%, more preferably at least
20%, more preferably at least 50% of the total material (by volume,
by wet or dry weight, or by mole percent or mole fraction) in a
sample is the protein or polypeptide of interest.
[0101] The invention should also be construed to include DNAs which
are substantially homologous to the DNA isolated according to the
method of the invention. Preferably, DNA which is substantially
homologous is about 50% homologous, more preferably about 70%
homologous, even more preferably about 80% homologous and most
preferably about 90% homologous to DNA obtained using the method of
the invention.
[0102] "Homologous" as used herein, refers to the subunit sequence
similarity between two polymeric molecules, e.g., between two
nucleic acid molecules, e.g., two DNA molecules or two RNA
molecules, or between two polypeptide molecules. When a subunit
position in both of the two molecules is occupied by the same
monomeric subunit, e.g., if a position in each of two DNA molecules
is occupied by adenine, then they are homologous at that position.
The homology between two sequences is a direct function of the
number of matching or homologous positions, e.g., if half (e.g.,
five positions in a polymer ten subunits in length) of the
positions in two compound sequences are homologous then the two
sequences are 50% homologous, if 90% of the positions, e.g., 9 of
10, are matched or homologous, the two sequences share 90%
homology. By way of example, the DNA sequences 3' ATTGCC 5' and 3'
TATGCG 5' share 50% homology.
[0103] To obtain a substantially pure preparation of a protein
comprising, for example, an antibody, generated using the methods
of the invention, the protein may be extracted from the surface of
the phage on which it is expressed. The procedures for such
extraction are well known to those in the art of protein
purification. Alternatively, a substantially pure preparation of a
protein comprising, for example, an antibody, may be obtained by
cloning an isolated DNA encoding the antibody into an expression
vector and expressing the protein therefrom. Protein so expressed
may be obtained using ordinary protein purification procedures well
known in the art.
[0104] As used herein, the term "substantially pure" describes a
compound, e.g., a protein or polypeptide which has been separated
from components which naturally accompany it. Typically, a compound
is substantially pure when at least 10%, more preferably at least
20%, more preferably at least 50%, more preferably at least 60%,
more preferably at least 75%, more preferably at least 90%, and
most preferably at least 99% of the total material (by volume, by
wet or dry weight, or by mole percent or mole fraction) in a sample
is the compound of interest. Purity can be measured by any
appropriate method, e.g., in the case of polypeptides by column
chromatography, gel electrophoresis or HPLC analysis. A compound,
e.g., a protein, is also substantially purified when it is
essentially free of naturally associated components or when it is
separated from the native contaminants which accompany it in its
natural state.
[0105] As used herein, amino acids are represented by the full name
thereof, by the three letter code corresponding thereto, or by the
one-letter code corresponding thereto, as indicated in the
following table: TABLE-US-00001 Full Name Three-Letter Code
One-Letter Code Aspartic Acid Asp D Glutamic Acid Glu E Lysine Lys
K Arginine Arg R Histidine His H Tyrosine Tyr Y Cysteine Cys C
Asparagine Asn N Glutamine Gln Q Serine Ser S Threonine Thr T
Glycine Gly G Alanine Ala A Valine Val V Leucine Leu L Isoleucine
Ile I Methionine Met M Proline Pro P Phenylalanine Phe F Tryptophan
Trp W
[0106] The present invention also provides for analogs of proteins
or peptides obtained according to the methods of the invention.
Analogs can differ from naturally occurring proteins or peptides by
conservative amino acid sequence differences or by modifications
which do not affect sequence, or by both.
[0107] For example, conservative amino acid changes may be made,
which although they alter the primary sequence of the protein or
peptide, do not normally alter its function. Conservative amino
acid substitutions typically include substitutions within the
following groups:
[0108] glycine, alanine;
[0109] valine, isoleucine, leucine;
[0110] aspartic acid, glutamic acid;
[0111] asparagine, glutamine;
[0112] serine, threonine;
[0113] lysine, arginine;
[0114] phenylalanine, tyrosine.
[0115] Modifications (which do not normally alter primary sequence)
include in vivo, or in vitro chemical derivatization of
polypeptides, e.g., acetylation, or carboxylation. Also included
are modifications of glycosylation, e.g., those made by modifying
the glycosylation patterns of a polypeptide during its synthesis
and processing or in further processing steps; e.g., by exposing
the polypeptide to enzymes which affect glycosylation, e.g.,
mammalian glycosylating or deglycosylating enzymes. Also embraced
are sequences which have phosphorylated amino acid residues, e.g.,
phosphotyrosine, phosphoserine, or phosphothreonine.
[0116] Also included in the invention are polypeptides which have
been modified using ordinary molecular biological techniques so as
to improve their resistance to proteolytic degradation or to
optimize solubility properties. Analogs of such polypeptides
include those containing residues other than naturally occurring
L-amino acids, e.g., D-amino acids or non-naturally occurring
synthetic amino acids. The peptides of the invention are not
limited to products of any of the specific exemplary processes
listed herein.
[0117] In addition to substantially full length polypeptides, the
present invention provides for active fragments of the
polypeptides. A specific polypeptide is considered to be active if
it binds to an antigen-bearing moiety, for example, if a fragment
of an antibody binds to its corresponding antigen in the same
manner as the full length protein.
[0118] As used herein, the term "fragment," as applied to a
polypeptide, will ordinarily be at least about fifty contiguous
amino acids, typically at least about one hundred contiguous amino
acids, more typically at least about two hundred continuous amino
acids and usually at least about three hundred contiguous amino
acids in length.
[0119] The invention is further described in detail by reference to
the following experimental examples. These examples are provided
for purposes of illustration only, and are not intended to be
limiting unless otherwise specified. Thus, the invention should in
no way be construed as being limited to the following examples, but
rather, should be construed to encompass any and all variations
which become evident as a result of the teaching provided
herein.
EXAMPLE 1
Isolation of Cell Surface-Specific Human Monoclonal Antibodies
Using Phage Display and Magnetically-Activated Cell Sorting
[0120] The experiments described in this Example provide procedures
and results for the isolation and production of anti-Rh(D) red
blood cell antibodies using Fab/phage display.
[0121] A method is described in FIG. 1 for the isolation of
filamentous phage-displayed human monoclonal antibodies specific
for non-purifiable cell surface expressed molecules. To optimize
the capture of antigen-specific phage and minimize the binding of
irrelevant phage antibodies, a simultaneous positive and negative
selection strategy was employed. Cells bearing the antigen of
interest are pre-coated with magnetic beads and are diluted into an
excess of unmodified antigen-negative cells. Following incubation
of the cell admixture with a Fab/phage library, the antigen
positive cell population is retrieved using magnetically-activated
cell sorting, and antigen-specific Fab/phage are eluted and
propagated in bacterial culture. When this protocol was used with
magnetically-labeled (Rh(D)-positive and excess non-labeled
Rh(D)-negative human red blood cells and a Fab/phage library
constructed from human peripheral blood lymphocytes, dozens of
unique, clinically useful .sub..gamma.1.kappa. and
.sub..gamma.1.lamda. anti-Rh(D) antibodies were isolated from a
single alloimmunized individual.
[0122] The cell-surface selection method of the present invention
is readily adaptable for use in other systems, such as for the
identification of putative tumor-specific antigens, and provides a
rapid (less than one month), high yield approach for isolating
self-replicative antibody reagents directed at novel or
conformationally-dependent cell-surface epitopes.
[0123] Creation of Fab/Phage Display Libraries
[0124] Separate .sub..gamma.1.kappa. and .sub..gamma.1.lamda. phage
libraries were constructed from 2.times.10.sup.7 mononuclear cells
derived from the peripheral blood from an Rh(D)-negative individual
previously hyperimmunized with Rh(D)-positive red blood cells
(RBCs). The phagemid vector pComb3 (Barbas, 1991, Proc. Natl. Acad.
Sci. USA 88:7978-7982) was used to create the libraries utilizing
previously published methods (Barbas et al., 1991, Combinatorial
immunoglobulin libraries on the surface of phage (Phabs): Rapid
selection of antigen-specific Fabs. Methods: A Companion to Methods
in Enzymology 2:119-124; Siegel et al., 1994, Blood
83:2334-2344).
[0125] Briefly, cDNA was prepared from the mRNA of the donor cells
and heavy chain and light chain immunoglobulin (Ig) cDNA segments
were amplified using the polymerase chain reaction (PCR) and the
battery of human Ig primers described by Kang et al. (1991,
"Combinatorial Immunoglobulin Libraries on the Surface of Phage
(Phabs): Rapid Selection of Antigen-Specific Fabs. Methods: A
Companion to Methods" in Enzymology 2:111-118) supplemented by
those of Silverman et al. (1995, J. Clin. Invest. 96:417426). The
heavy and light chain PCR products were cloned into pComb3 and
electroporated into E. coli. Upon co-infection with VCSM13 helper
phage (Stratagene, La Jolla, Calif.), Ig DNA was packaged into
filamentous phage particles which express human Fab molecules fused
to the gene III bacteriophage coat protein.
[0126] Panning Fab Phage Display Libraries for Anti-Rh(D)
Clones
[0127] Rh(D)-positive RBCs were cell-surfaced biotinylated by
incubating cells at a hematocrit of 10% with 500 .mu.g/ml
sulfo-NHS-LC-biotin (Pierce Chemical, Rockford, Ill.) for 40
minutes at room temperature (RT). Following 5 washes with
phosphate-buffered saline (PBS), 8.times.10.sup.6 biotinylated
Rh(D)-positive RBCs were incubated with 10 .mu.l of
streptavidin-coated paramagnetic microbeads (MACS Streptavidin
Microbeads, Mitenyi Biotec, Sunnyvale, Calif.) for 1 hour at RT in
a total volume of 100 .mu.l PBS. Non-reacted beads were removed by
washing and then the magnetic bead-coated, Rh(D)-positive RBCs were
mixed with a 10-fold excess {8.times.10.sup.7) of the Rh(D)negative
(unmodified) RBCs and .about.3.times.10.sup.11 colony-forming units
(cfu) of either the .sub..lamda.1.kappa. and .sub..gamma.1.lamda.
Fab/phage libraries (prepared as described above) in a final volume
of 40 .mu.l PBS containing 2% non-fat dry milk (MPBS, Carnation,
Nestle Food Products, Glendale, Calif.).
[0128] Following a 2 hour incubation at 37.degree. C., the
RBC/phage suspension was loaded at a flow rate of 10 .mu.l/minute
onto a MiniMACS magnetic type MS column (Mitenyi Biotec, Sunnyvale,
Calif.) that was pre-equilibrated with 2% MPBS. This loading step
was performed without a magnetic field around the column so as to
prevent magnetic bead-coated RBCs from instantly adhering to the
very top of the column, clogging it, and causing the trapping of
Rh(D)negative non-biotinylated RBCs. Loading the RBC/phage
incubation mixture in the absence of a magnetic field causes the
antigen-negative and antigen-positive RBCs to distribute evenly
throughout the column without running off since the excluded volume
of the column is slightly greater than 40 .mu.l. Once loaded, the
column was placed in a magnetic field (MiniMACS magnetic separation
unit, Mitenyi Biotec, Sunnyvale, Calif.) for 2 minutes to allow the
Rh(D)-positive RBCs to adhere, and a series of 500 .mu.l washes
were performed with ice-cold MPBS followed by a final wash with
PBS. A total of 3 washes were performed for the first 2 rounds of
panning and a total of 6 washes were performed for all subsequent
pannings. For each panning, the first wash was carried out at a
flow rate of 10 .mu.l/minute during which time the bulk of
Rh(D)-negative RBCs washed off the column. All subsequent washes
were performed at 200 .mu.l/minute. Following the last wash, the
column was removed from the magnetic field and the
bead-coated/phage-coated Rh(D)-positive RBCs were flushed off the
column with 500 .mu.l PBS using the plunger from a 5 cc syringe
(Becton-Dickinson, Franklin Lakes, N.J.).
[0129] The RBCs were immediately centrifuged for 5 seconds at
13,000.times. g and were then resuspended in 200 .mu.l of 76 mM
citrate, pH 2.4, to denature the Rh(D) antigen and elute bound
phage. Following a 10 minute incubation period at RT with
intermittent vortexing, the phage eluate and cellular debris were
neutralized with 18 .mu.l 2 M Tris base and were added to 10 ml of
O.D.=1.0 XL1-Blue strain of E. coli (Stratagene, La Jolla, Calif.)
grown in super broth (SB) (Barbas et al., 1991, supra) supplemented
with 10 .mu.g/ml tetracycline. After incubation for 15 minutes at
RT, during which time the phage library enriched for Rh(D) binders
was allowed to infect the bacterial culture, 10 ml of pre-warmed,
37.degree. C. SB containing 40 .mu.g/ml carbenicillin/10 .mu.g/mil
tetracycline was added to give final antibiotic concentrations of
20 .mu.g/ml and 10 .mu.g/ml, respectively. A small aliquot of
culture (.about.100 .mu.l) was immediately removed and titered on
Luria broth/carbeniclllin plates to determine the number of phage
contained in the total eluate. The balance of the culture was
shaken at 37.degree. C. for 1 hour at 300 RPM. Additional
antibiotics, additional SB, and VCSM13 helper phage were
subsequently added and the culture was grown overnight at
30.degree. C. as described (Siegel et al., 1994, supra).
[0130] Phagemid particles were purified from the culture
supernatant by polyethylene glycol 8000 (PEG) precipitation (Barbas
et al., 1991, supra), resuspended in 1% bovine serum albumin
(BSA)/PBS, and dialyzed overnight to remove residual PEG that may
lyse RBCs during subsequent rounds of panning. Thus, the resultant
phage preparation serves as the input for the next round of
panning. The .sub..gamma.1.kappa. and .sub..gamma.1.lamda. phage
libraries were panned separately to prevent any bias in light chain
isotype replication possibly introduced by bacterial
amplification.
[0131] Screening Polyclonal Fab/Phage Libraries and Individual
Phage Colonies for Anti-Rh(D) Reactivity
[0132] The specificity of Fab/phage for the Rh(D) antigen was
assessed using anti-M13 antibody as a bridging antibody to induce
agglutination between RBCs that have bound anti-Rh(D) Fab/phage.
One hundred .mu.l aliquots of polyclonal Fab/phage from rounds of
panning, or monoclonal Fab/phage derived from individual Fab/phage
eluate clones, were incubated with 50 .mu.l of a 3% suspension of
RBCs of defined phenotype (ie., Rh(D)-negative or -positive).
[0133] Following 1 hour incubation at 37.degree. C., the RBCS were
washed 3 times with 2 ml cold PBS to remove unbound Fab/phage. The
resultant RBC pellets were resuspended in 100 .mu.l of a 10
.mu.g/ml solution of sheep anti-M13 antibody (5-Prime 3-Prime,
Boulder, Colo.) and transferred to the round-bottomed wells of a
96-well microtiter plate. Plates were left undisturbed (.about.2
hours) and were then read. Wells having a negative reaction exhibit
sharp .about.2 millimeter diameter RBC spots whereas in wells
having positive reactions, i.e., agglutination, the RBCs in
agglutinated wells form a thin carpet coating the entire floor of
the well.
[0134] For hemagglutination assays utilizing mini-column gel cards
(ID-Micro-Typing System, Ortho Diagnostics, Raritan, N.J.)
(Lapierre et al., 1990, Transfusion 30:109-113), 25 .mu.l of
Fab/phage clones were mixed with 50 .mu.l aliquots of RBCs (0.8%
suspensions in Micro Typing System buffer, Ortho Diagnostics). The
mixtures were placed in the reservoirs above the mini-columns which
contain dextran-acrylamide beads previously suspended in 100
.mu.l/ml anti-M13 antibody. After incubation at 37.degree. C., the
gel cards were centrifuged at 70.times. g for 10 minutes and were
read.
[0135] Miscellaneous Methods
[0136] Preparation of fluorescently-labeled RBCs for flow cytometry
was performed as described herein and samples were analyzed using a
FACScan microfluorimeter equipped with Lysis II (Ver 1.1) software
(Becton-Dickinson, Mountain View, Calif.). Plasmid DNA was prepared
from bacterial clones (Qiawell Plus, Qiagen, Chatsworth, Calif.).
Double-stranded DNA was sequenced using light chain or heavy chain
Ig constant region reverse primers or unique pComb3 vector primers
that anneal 5-prime to the respective Ig chain (Barbas et al.,
1991, supra; Roben et al., 1995, J. Immunol. 154:6437-6445) and
automated fluorescence sequencing (Applied Biosystems, Foster City,
Calif.). Sequences were analyzed using MacVector Version 5.0
sequencing software (Oxford Molecular Group, Oxford, UK) and the
Tomlinson database of Ig germline genes (Tomlinson et al., 1996, V
Base Sequence Directory. MRC Center for Protein Engineering,
Cambridge, UK).
[0137] Experimental design for cell incubation and separation
protocols The experimental conditions described above for panning
Fab/phage libraries for anti-RBC-reactive phage were determined
after performing a series of initial studies aimed at optimizing
the cell separation process and ultimate yield of antigen-specific
Fab/phage. The main parameters investigated included:
[0138] Biotinylation Conditions were sought that would biotinylate
the RBC surface in a manner such that a sufficient number of
streptavidin-coated magnetic beads would bind to the cells causing
the RBCs to be retained by a magnetic column. In this case,
over-biotinylation that might destroy the antigenicity of the Rh(D)
antigen or might make the cells non-specifically absorb antibody is
to be avoided. To address this issue, Rh(D)-positive/Kell-negative
RBCs (Kell being a RBC antigen; (Walker, ed. 1993, In: Technical
Manual 11.sup.th Ed., Bethesda, Md., American Association of Blood
Banks) were incubated with a range of sulfo-NHS-LC-biotin
concentrations and the degree of biotinylation was assessed by flow
cytometry utilizing fluorescein-conjugated streptavidin.
[0139] To assess the degree of cell-surface biotinylation, 5 .mu.l
aliquots of 3% suspensions of Rh(D)-positive/Kell-negative RBCs
biotinylated at varying biotin reagent concentrations were
incubated with 200 .mu.l of a 1/100 dilution of FITC-streptavidin
(Jackson ImmunoResearch, Bar Harbor, Me.) for 30 min at 4.degree.
C. (FIG. 2). The mixture was washed with phosphate buffered saline
(PBS) and analyzed by flow microfluorimetry (-.quadrature.-).
Aliquots of cells were also analyzed for retention of
Rh(D)-antigenicity (-.DELTA.-) (i.e., specific staining) or for
lack of non-specific staining (-.largecircle.-) by incubating the
cells with 100 .mu.l of either anti-Rh(D) or anti-Kell typing sera,
respectively, washing the cells and then staining them with a 1/100
dilution of FITC-goat anti-human IgG (Jackson ImmunoResearch).
[0140] A linear, non-saturating response was observed (FIG. 2).
Retention of Rh(D) antigenicity was assessed using anti-Rh(D)
typing serum and was found to be unaffected by the derivatization
of cell-surface proteins with biotin at all biotin concentrations
tested (FIG. 2). Furthermore, the Kell-negative RBCs did not
non-specifically adsorb anti-Kell antibodies.
[0141] Each biotinylated RBC sample was then incubated with an
excess of streptavidin-coated magnetic microbeads and applied to a
magnetic separation column. It was determined that as many as 108
RBCs could be retained by the column for RBC samples biotinylated
with greater than or equal to 500 .mu.g/ml biotin reagent. Since
the actual RBC/phage panning experiments were designed to use only
.about.10.sup.7 Rh(D)-positive cells (see below), RBC biotinylation
at 500 .mu.g/ml was determined to be sufficient.
[0142] Concentration of Rh(D)-Positive and Rh(D)-Negative RBCs in
Incubation Mixture
[0143] Prior to performing Fab/phage panning experiments, the
ability of the magnetically-activated cell separation technique to
separate Rh(D)-positive and Rh(D)-negative cells was assessed using
anti-Rh(D) typing serum and flow cytometry (FIG. 3).
Streptavidin-microbead coated, biotinylated Rh(D)-positive RBCs
(8.times.10.sup.6 cells) were mixed with a 10-fold excess of
Rh(D)-negative non-coated RBCs (8.times.10.sup.7 cells) in a 40
.mu.l volume of PBS containing 2% non-fat dry milk (MPBS) and the
mixture was applied to a MiniMACS column. The column was washed and
the bound cells were eluted as described herein. Aliquots of RBCs
contained in the original admixture (panel a), the column wash
(panel b), and the column eluate (panel c) were stained with
anti-Rh(D) typing serum and FITC-goat anti-human IgG as described
in FIG. 2. The flow cytograms show that although -90% of the cells
in the column load were Rh(D)-negative (panel a), nearly all of
them washed off of the column (panel b), yielding a column eluate
that was almost entirely Rh(D)-positive cells (panel c). Since only
.about.6% of the final eluate comprise Rh(D)-negative cells (panel
c), and Rh(D)-negative cells were initially present in a 10-fold
excess to Rh(D)-positive cells, only 0.6% of the initial
antigen-negative immunosorbent cells contaminated the final
antigen-positive preparation. This efficiency of the cell
separation was deemed adequate for subsequent panning experiments
with Fab/phage.
[0144] In the above-described experiment, to avoid clogging the
magnetic separation column, it was necessary to load the column in
the absence of a magnetic field. This necessitated a reaction
volume of less than or equal to 40 .mu.l so that none of the
material would run off the column. On theoretical grounds
(Kretzschmar et al., 1995, Anal. Biochem. 224:413419), one can
calculate the appropriate concentration of cells required in a 40
.mu.l volume to capture greater than 50% of Fab/phage specific for
a given cell surface antigen. Such a calculation is a function of
the number of antigen sites per cell and the dissociation constant
(K.sub.D) of the bound Fab/phage. Using a value of .about.100,000
Rh(D) antigen sites per RBC (phenotype "-D-/-D-") (Mollison et al.,
1993, In: Blood Transfusion in Clinical Medicine," Oxford,
Blackwell Scientific Publications) and the desired Fab/phage
affinity in the K.sub.D=10.sup.-8 to 10.sup.-9 M range, then
8.times.10.sup.6 Rh(D)-positive RBCs in a 40 .mu.l reaction volume
would be required. Given this number of Rh(D)-positive cells, a
10-fold excess of Rh(D)-negative RBCs was found to be the maximum
amount of antigen-negative cells that could be effectively
separated from antigen-positive RBCs by the magnetic column (FIG.
3).
[0145] Construction and Panning of Fab/Phage Libraries
[0146] .sub..gamma.1.kappa. and .sub..gamma.1.lamda. phage
libraries were prepared as described herein and were found to
contain 7.times.10.sup.7and 3.times.10.sup.8 independent
transformants, respectively. Table 1 tabulates the panning results
for the libraries.
[0147] An RBC agglutination assay utilizing anti-M13 secondary
antibody as bridging antibody was used to detect anti-Rh(D)
Fab/phage activity in the panned polyclonal libraries and the
individual randomly-picked Fab/phage clones (FIG. 4). The results
shown are a representative example of the assay depicting negative
reactivity to Rh(D)-negative RBCs and strongly positive reactivity
to Rh(D)-positive RBCs for the .sub..gamma.1.kappa. library
(panning #2) out to a dilution of 1/2048.
[0148] In the case of the .sub..gamma.1.kappa. library, significant
enrichment for binding phage appears to occur after only one round
of panning, whereas significant enrichment for the
.sub..gamma.1.lamda. , library occurs during the second round. This
is reflected by both the sharp increase in the percent of phage
bound during a given round of panning as well as the ability of the
polyclonal .sub..gamma.1.kappa. and .sub..gamma.1.lamda. Fab/phage
libraries to agglutinate Rh(D)-positive RBCs after 1 and 2 rounds
of panning, respectively (Table 1, FIG. 4).
[0149] Monoclonal Fab/phage were prepared from randomly-picked
individual bacterial colonies obtained during each round of
panning. It was apparent that by the third round of panning, all
clones have anti-Rh(D) specificity (Table 1). To confirm that these
Fab/phage have anti-Rh(D) specificity and are not binding to other
unrelated antigens that may coincidentally be present on the
particular Rh(D)-positive RBC and absent on the particular
Rh(D)-negative RBC used in the agglutination assays, clones were
screened against a panel of 11 Rh(D)negative and-positive RBCs of
varying blood group specificities to verify their anti-Rh(D)
specificity (Walker, 1993, supra).
[0150] Clonal Analysis at the Genetic Level
[0151] To investigate the genetic diversity among the randomly
picked anti-Rh(D) clones, plasmid DNA was prepared from each of the
clones and the corresponding heavy and light chain Ig nucleotide
sequences were identified. In Table 2 there is listed a number of
attributes for each clone including the name of the most
closely-related germline heavy or light chain Ig gene. More
detailed analysis at the nucleotide level revealed that among all
of the anti-Rh(D) binding clones, there were a large number of
unique heavy and light chain DNA sequences (Table 3). Because of
the random shuffling of heavy and light chain gene segments which
occurs during the creation of a Fab/phage display library (Barbas
et al., 1991, supra), it is evident that these heavy chains and
light chains combined to form nearly 50 different anti-Rh(D)
antibodies.
[0152] A detailed multiple alignment analysis of the predicted
amino acid sequences revealed a total of twenty-five unique heavy
chain, eighteen unique kappa light chain and twenty-three unique
lambda light chain proteins. Due to the combinatorial effect during
library construction, these heavy and light chain gene segments
paired to produce fifty unique Fab antibodies
(20.sub..gamma.1.kappa. and 30.sub..gamma.1.lamda.). Of interest,
all twenty five unique heavy chains and nearly all of the eighteen
unique kappa light chains were derived from only 5 V.sub.HIII or
four V.kappa.I germline genes, respectively, while the lambda light
chains were derived from a more diverse set of germline genes.
Analysis of heavy and light chain nucleotide sequences from over
sixty negative clones from the non-panned libraries were performed
to verify the heterogeneity in variable region family
representation before selection. Clones representing V.sub.H
families I (13%), III (36%), IV (31%), V(15%) and VI (5%); V.kappa.
families I (43%), II (14%), III (29%) and IV (14%); and V.gamma.
families I (48%), II (4%), III (9%), IV (4%), V (9%), VI (17%) and
VII (9%) were present.
[0153] Clonal Analysis at the Protein Level
[0154] To investigate the diversity in fine specificity (Rh(D)
antigen epitope specificity) among the anti-Rh(D) clones,
agglutination experiments were performed with selected clones and
with sets of rare Rh(D)-positive RBCs which were obtained from
individuals whose RBCs produce Rh(D) antigen lacking certain
epitopes. Examining the pattern of agglutination of a particular
anti-Rh(D) antibody with such sets of mutant RBCs enables the
identification of the specific epitope on Rh(D) to which the
antibody is directed (Mollison et al., 1993, supra). A
representative example of such an experiment is shown in FIG. 5 and
the Rh(D) epitopes for selected anti-Rh(D) Fab/phage clones are
tabulated in Table 2.
[0155] Agglutination experiments were performed with
anti-Rh(D)-negative RBCs (rr), Rh(D)-positive RBCs
(R.sub.2R.sub.2), and "partial" Rh(D)-positive RBCs (mosaics IIIa,
IVa, Va, VI, VII). The results shown are a representative example
of the assay for 5 randomly-picked anti-Rh(D) Fab/phage clones
(FIG. 5). TABLE-US-00002 TABLE 1a .sub..gamma.1.kappa.FAB/PHAGE
LIBRARY PANNING RESULTS .phi.INPUT .phi.OUTPUT % BOUND.sup.4 AGGLU
T BINDERS/ PANNING.sup.1 (CFUs).sup.2 (CFUs).sup.3
(.times.10.sup.-4) ENRICHMENT.sup.5 TITER.sup.6 TOTAL(%).sup.7 0 0
0/16 (0) 1 2.94 .times. 10.sup.11 6.04 .times. 10.sup.5 2.1 1/16
0/16 (0) 2 2.15 .times. 10.sup.11 1.68 .times. 10.sup.7 78.3
38.0.times. 1/2048 15/15 (100) 3 1.72 .times. 10.sup.11 1.44
.times. 10.sup.8 840.0 10.7.times. 1/2048 12/12 (100)
[0156] TABLE-US-00003 TABLE 1b .sub..gamma.1.lamda.FAB/PHAGE
LIBRARY PANNING RESULTS .phi.INPUT .phi.OUTPUT % BOUND.sup.4 AGGLU
T BINDERS/ PANNING.sup.1 (CFUs).sup.2 (CFUs).sup.3
(.times.10.sup.-4) ENRICHMENT.sup.5 TITER.sup.6 TOTAL(%).sup.7 0 0
0/16 (0) 1 2.28 .times. 10.sup.11 3.48 .times. 10.sup.5 1.5 0 2
5.51 .times. 10.sup.11 1.34 .times. 10.sup.6 2.4 1.6.times. 1/128
32/36 (89) 3 3.93 .times. 10.sup.11 3.86 .times. 10.sup.8 980.0
404.0.times. 1/512 24/24 (100) 4 2.87 .times. 10.sup.11 3.08
.times. 10.sup.8 1100.0 1.1.times. 1/1024 .sup.1panning round,
where "0" represents the initial, non-panned Fab/phage library
.sup.2number of colony-forming units (CFUs) of phage (.phi.)
incubated with Rh(D)-positive/-negative RBC admixture .sup.3total
number of CFUs of .phi. contained in eluate .sup.4(.phi.
output/.phi.input) .times. 100 .sup.5fold increase in % bound from
compared to previous round of panning .sup.6agglutination titer;
see text and FIG. 4 .sup.7number of Rh(D)-binding Fab/phage clones
per total number of clones screened from panning round; see Table 2
for details
[0157] TABLE-US-00004 TABLE 2a ANALYSIS OF
.sub..gamma.1.kappa.FAB/PHAGE CLONES AGGL VH V.kappa. CLONE.sup.1
U.sup.2 FAM.sup.3 VH GENE.sup.4 FAM.sup.5 V.kappa. GENE.sup.6 D
EPITOPE.sup.7 KPO-1 neg 3 DP-47/V3-23 4 DPK24/VklVKlobeck KPO-2 neg
3 DP-31/V3-9P 3 DPK22/A27 KPO-3 neg 3 DP-58/hv3d1EG 4
DPK24/VklVKlobeck KPO-4 neg 4 3d279d+ -- no light chain KPO-5 neg 3
DP-29/12-2 1 LFVK431 KPO-6 neg 4 DP-79/4d154 1 DPK9/012 KPO-7 neg 3
V3-48/hv3d1 4 DPK24/VklVKlobeck KPO-8 neg 4 DP-70/4d68 2 DPK18/A17
KPO-9 neg 1 DP-14/V1-18 1 DPK9/012 KPO-10 neg 4 DP-70/4d68 1
DPK9/012 KPO-11 neg 5 DP-73/V5-51 1 DPK9/012 KPO-12 neg 3
DP-54/V3-7 2 DPK18/A17 KPO-13 neg 3 V3-48/hv3d1 1 Vb' KPO-14 neg 6
DP-74/VH-VI 1 DPK6/Vb'' KPO-15 neg 3 DP-46/3d216 3 Vg/38K KPO-16
neg 6 DP-74/VH-VI 1 DPK9/012 KP1-1 neg 4 V71-4+ 3 DPK22/A27 KP1-2
neg 4 3d279d+ 1 DPK8/Vd+ KP1-3 neg 1 4M28 1 DPK9/012 KP1-4 neg 4
DP-79/4d154 3 Vg/38K KP1-5 neg 3 DP-38/9-1 3 DPK22/A27 KP1-6 neg 4
DP-70/4d68 1 L12a/PCRdil6-5 KP1-7 neg 5 DP-73/V5-51 2 DPK15/A19
KP1-8 neg 4 DP-70/4d68 3 DPK22/A27 KP1-9 neg -- no heavy chain --
no light chain KP1-10 neg -- no heavy chain 3 DPK22/A27 KP1-11 neg
1 DP-15/V1-8+ 1 DPK9/012 KP1-12 neg 3 b28e -- no light chain KP1-13
neg 3 DP-47/V3-23 4 DPK24/VklVKlobeck KP1-14 neg 3 DP-31/V3-9P 3
DPK21/humkv328h5 KP1-15 neg 1 DP-7/21-2 4 DPK24/VklVKlobeck KP1-16
neg 5 DP-73/V51 3 DPK22/A27 KP2-1 pos 3 DP-50/hv3019b9 1 DPK9/012
epD6/7 KP2-2 pos 3 DP-50/hv3019b9 1 DPK9/012 epD6/7 KP2-3 pos 3
DP-50/hv3019b9 1 DPK9/012 epD6/7 KP2-4 pos 3 b28m 1 DPK9/012 epD2
KP2-5 pos 3 b28m 1 DPK9/012 epD1 KP2-6 pos 3 DP-50/hv3019b9 1
DPK9/012 epD6/7 KP2-7 pos 3 DP-50/hv3019b9 1 DPK9/012 epD5 KP2-8
pos 3 DP-50/hv3019b9 1 DPK9/012 KP2-9 pos 3 DP-50/hv3019b9 1
DPK9/012 epD2 KP2-10 pos 3 DP-50/hv3019b9 1 DPK9/012 epD2 KP2-11
pos 3 DP-50/hv3019b9 1 DPK9/012 epD2 KP2-12 pos 3 DP-50/hv3019b9 1
DPK9/012 epD1 KP2-13 pos 3 DP-50/hv3019b9 1 DPK9/012 epD6/7 KP2-14
pos 3 DP-50/hv3019b9 2 DPK15/A19 epD2 KP2-15 pos 3 DP-50/hv3019b9 1
DPK9/012 epD6/7 KP3-1 pos 3 DP-50/hv3019b9 1 DPK9/012 KP3-2 pos 3
DP-50/hv3019b9 1 DPK9/012 epD6/7 KP3-3 pos 3 DP-50/hv3019b9 1
DPK9/012 KP3-4 pos 3 DP-49/1.9111 1 DPK9/012 epD5 KP3-5 pos 3
DP-50/hv3019b9 1 DPK9/012 KP3-6 pos 3 DP-50/hv3019b9 1 A30/SG3+
epD6/7 KP3-7 pos 3 DP-50/hv3019b9 1 DPK8/Vd+ epD6/7 KP3-8 pos 3
DP-50/hv3019b9 1 DPK9/012 epD6/7 KP3-9 pos 3 DP-50/hv3019b9 1
DPK9/012 KP3-1O pos 3 DP-50/hv3019b9 1 DPK9/012 KP3-11 pos 3
DP-50/hv3019b9 1 DPK9/012 KP3-12 pos 3 DP-46/3d216 1 DPK9/012
.sup.1nomenclature: prefix "KPO" denotes
".sub..gamma.1.kappa.Fab/phage library, panning 0", "KP1" denotes
".sub..gamma.1.kappa.Fab/phage library, panning 1", etc.
.sup.2agglutination negative or positive against Rh(D)-positive RBC
.sup.3Ig heavy chain variable region gene family per Tomlinson et
al., supra .sup.4closest related Ig heavy chain variable region
gene per Tomlinson et al. supra .sup.5Ig light chain variable
region gene family per Tomlinson et al., supra .sup.6closest
related Ig light chain variable region gene per Tomlinson et al.,
supra .sup.7Rh(D) epitope as defined by rare RBC agglutination
pattern (see FIG. 5 and text)
[0158] TABLE-US-00005 TABLE 2b ANALYSIS OF
.sub..gamma.1.lamda.FAB/PHAGE CLONES AGGL VH V.kappa. CLONE.sup.1
U.sup.2 FAM.sup.3 VH GENE.sup.4 FAM.sup.5 V.kappa. GENE.sup.6 D
EPITOPE.sup.7 LPO-1 neg 4 DP-65/3d75d 1 DPL7/IGLV1S2 LPO-4 neg 4
DP-70/4d68 6 IGLV8A1 LPO-3 neg 6 DP-74/VH-VI 7 DPL18/VL7.1 LPO-4
neg 3 DP-29/12-2 1 DPL3/Iv122 LPO-5 neg 3 DP-38/9-1 6
IGLV6S1/LV6SW-G LPO-6 neg 1 4M28 1 DPL3/Iv122 LPO-7 neg 1 8M27 1
DPL2/Iv1L1 LPO-8 neg 5 DP-58/V5-51 6 IGLV6S1/LV6SW-G LPO-9 neg 5
DP-73/V5-51 1 DPL7/IGLV1S2 LPO-10 neg 3 DP-38/9-1 1 DPL2/Iv1L1
LPO-11 neg 3 DP-31/V3-9P 3 DPL23/VLIII.1 LPO-12 neg -- no heavy
chain 1 DPL7/IGLV1S2 LPO-13 neg 3 DP-47/V3-23 -- no light chain
LPO-14 neg 4 DP-71/3d197d 6 IGLV6S1/LV6SW-G LPO-15 neg 4 DP-70/4d68
4 IGLV8A1 LPO-16 neg 3 DP-54/V3-7 7 DPL19 LP2-1 pos 3
DP-50/hv3019b9 1 DPL2/Iv1L1 epD2 LP2-2 pos 3 DP-77/WHG16 1
DPL3/Iv122 LP2-3 pos 3 DP-49/1.9111 1 DPL3/Iv122 epD1 LP2-4 neg 4
3d279d+ 1 DPL2/Iv1L1 LP2-5 pos 3 DP-49/1.9111 3 DPL16/IGLV3S1 epD5
LP2-6 pos 3 DP-50/hv3019b9 1 DPL7/IGLV1S2 epd2 LP2-7 pos 3 b28m 1
DPL7/IGLV1S2 epD2 LP2-8 pos 3 DP-49/1.9111 3 IGLV3S2=Iv318 epD1
LP2-9 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 epD2 LP2-10 pos 3
DP-77/WHG16 1 DPL3/LV122 LP2-11 neg 1 DP-75-VI-2 1 DPL5/LV117d
LP2-12 pos 3 DP-77/WHG16 1 DPL2/LV1L1 epD2 LP2-13 pos 3
COS-8/hv3005f3 4 IGLV8A1 LP2-14 pos 3 DP-49/1.9111 1 DPL7/IGLV1S2
epD5 LP2-15 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 LP2-16 pos 3
DP-49/1.9111 2 Iv2046 epd1 LP2-17 pos 3 DP-77/WHG16=V3-21+ 1
DPL3/Iv122 epD3/9 LP2-18 pos 3 DP-49/1.9111 2
VL2.1.about.DPL10/Iv2066 epD1 LP2-19 pos 3 DP-50/hv3019b9 3
DPL16/IGLV3S1 epD2 LP2-20 neg 3 V3-49+ 3 DPL16/IGLV3S1 LP2-21 pos 3
DP-50/hv3019b9 7 DPL18/VL7.1 epD6/7 LP2-22 pos 3 DP-49/1.9111 2
Iv2046 LP2-23 pos 3 DP-49/1.9111 3 DPL16/IGLV3S1 epD5 LP2-24 pos 3
DP-77/WHG16 1 DPL3/Iv122 LP2-25 pos 3 DP-50/hv3019b9 7 DPL18/VL7.1
epD6/7 LP2-26 pos 3 DP-49/1.9111 3 DPL16/IGLV3S1 LP2-27 neg 3
COS-6/DA-8 2 VL2.1 LP2-28 pos 3 COS-8/hv3005f3 4 IGLV8A1 LP2-29 pos
3 DP-49/1.9111 DPL13 LP2-30 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1
LP2-31 pos 3 DP-50/hv3019b9 7 DPL18/VL7.1 LP2-32 pos 3 DP-49/1.9111
1 DPL2/Iv1L1 LP2-33 pos 3 DP-50/hv3019b9 7 DPL18/VL7.1 LP2-34 pos 3
DP-50/hv3019b9 7 DPL18/VL7.1 LP2-35 pos 3 DP-50/hv3019b9 3
DPL16/IGLV3S1 LP2-36 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 LP3-1 pos
3 DP-50/hv3019b9 3 DPL16/IGLV3S1 epD2 LP3-2 pos 3 DP-49/1.9111 3
DPL16/IGLV3S1 epD1 LP3-3 pos 3 DP-49/1.9111 3 DPL16/IGLV3S1 LP3-4
pos 3 DP-50/hv3019b9 7 DPL18/VL7.1 epD6/7 LP3-5 pos 3 DP-49/1.9111
1 DPL5/LV117d epD5 LP3-6 pos 3 DP-49/1.9111 1 DPL5/LV117d epD1
LP3-7 pos 3 DP-77/WHG16 1 DPL2/Iv1L1 epD5 LP3-8 pos 3 b28m 1
DPL7/IGLV1S2 epD2 LP3-9 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 epD2
LP3-1O pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 LP3-11 pos 3
DP-50/hv3019b9 3 DPL16/IGLV3S1 epD2 LP3-12 pos 3 COS-8/hv3005f3 4
IGLV8A1 epD6/7 LP3-13 pos 3 DP-50/hv3019b9 1 DPL2/Iv1L1 epD2 LP3-14
pos 3 DP-49/1.9111 3 DPL16/IGLV3S1 LP3-15 pos 3 DP-77/WHG16 1
DPL3/Iv122 epD1 LP3-16 pos 3 DP-49/1.9111 1 DPL2/Iv1L1 epD5 LP3-17
pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 LP3-18 pos 3 DP-50/hv3019b9 3
DPL16/IGLV3S1 LP3-19 pos 3 DP-50/hv3019b9 3 DPL16/IGLV3S1 epD5
LP3-20 pos 3 DP-50/hv3019b9 1 DPL2/Iv1L1 LP3-21 pos 3 DP-49/1.9111
1 DPL3/Iv122 LP3-22 pos 3 COS-8/hv3005f3 1 DPL2/Iv1L1 LP3-23 pos 3
DP-49/1.9111 3 DPL16/IGLV3S1 LP3-24 pos 3 DP-50/hv3019b9 3
DPL16/IGLV3S1 .sup.1nomenclature: prefix "LPO" denotes
".sub..gamma.1.lamda.Fab/phage library, panning 0", "LP1" denotes
".sub..gamma.1.lamda.Fab/phage library, panning 1", etc.
.sup.2agglutination negative or positive against Rh(D)-positive RBC
.sup.3Ig heavy chain variable region gene family per Tomlinson et
al., supra .sup.4closest related Ig heavy chain variable region
gene per Tomlinson et al., supra .sup.5Ig light chain variable
region gene family per Tomlinson et al., supra .sup.6closest
related Ig light chain variable region gene per Tomlinson et al.,
supra .sup.7Rh(D) epitope as defined by rare RBC agglutination
pattern (see FIG. 5 and text)
[0159] TABLE-US-00006 TABLE 3 SUMMARY OF FAB/PHAGE CLONAL ANALYSIS
Number of unique heavy chains 25 Number of unique .kappa. light
chains 18 Number of unique .lamda. light chains 23 Number of
.sub..gamma.1.kappa. antibodies 20 Number of .sub..gamma.1.lamda.
antibodies 30 Number Rh(D) epitope specificities represented 5
Use of Fab/Phage Antibodies as Blood Bank Typing Reagents
[0160] The ability of the anti-Rh(D) Fab/phage preparations to
accurately distinguish Rh(D)-negative from Rh(D)-positive RBCs in
microplate hemagglutination assays (FIGS. 4 and 5) provided
evidence that a gel test (Lapierre et al., 1990, Transfusion
30:109-1130) used by blood banks to phenotype RBCs using
conventional antisera could be adapted for use with Fab/phage.
[0161] The gel test comprises a plastic card of approximately
5.times.7 cm, containing 6 mini-columns each filed with about 20
.mu.l of dextran-acrylamide beads suspended in anti-human globulin
(Coombs reagent). Red cells to be typed are incubated with the
desired human anti-sera and are centrifuged through the gel. RBCs
which are positive for antigens to which the antisera is directed
agglutinate as they encounter the anti-human globulin and become
trapped in or above the gel matrix.
[0162] Unreactive RBCs sediment through the gel particles and form
a pellet at the bottom of the microtube. Because the gel test
offers a number of advantages over traditional blood banking
methods for RBC phenotyping including decreased reagent volumes,
the elimination of a cell washing step and a more objective
interpretation of results, many blood bank facilities have adapted
this new technology. As shown in FIG. 6, anti-Rh-(D) Fab/phage can
be used with gel cards that are modified to contain anti-M13
antibody. 2 0 To perform the assay, Rh(D)-negative or -positive red
blood cells were incubated with dilutions of anti-Rh(D) Fab/phage
(.gamma..sub.1.kappa. library, panning #2) and were centrifuged
into micro-columns containing beads suspended in anti-M13
antibody.
[0163] Undiluted Fab/phage stock had a titer of 5.times.10.sup.12
cfu/ml similar to that in the microplate settling assay (FIG. 4).
Because the volume of Fab/phage used in this assay is one-fourth of
that in the microplate assay, the amount of Fab/phage present in
the 1/625 dilution is approximately equal to that present in the
1/2048 dilution in FIG. 4. Therefore, the number of Fab/phage
required to yield a positive result is essentially equivalent in
both assays.
[0164] In other assays which were performed as just described, when
anti-M13 antibody was eliminated from the assay, no agglutination
of red blood cells was observed. In addition, anti-IgG antibody
does not react with recombinant Fabs expressed on the surface of
the bacteriophage. Only Rh-positive cells which were reacted with
anti-Rh phage were agglutinated when anti-M 13 antibody was present
in the assay. It should be noted that when high concentrations of
anti-M13 antibody were used, even Rh-negative cells appeared to be
agglutinated. This is an artifact resulting from the cross-linking
of unbound (i.e., non-reacted) phage which becomes crosslinked in
the presence of high amounts of anti-M13 antibody and forms a
semi-impenetrable mat through which not all the Rh-negative cells
can traverse. In the experiments described herein, an anti-M13
concentration of about 100 .mu.g/ml was considered to be optimal
for agglutination and for the prevention of false positive results.
Depending on the precise concentrations of reagents and cells used
in the assay, the concentration of anti-M13 may deviate from this
number.
[0165] To assess the relative sensitivity of an anti-M13 modified
Micro Typing System, the columns of the Micro Typing System cards
had added to them 100 .mu.g/ml of anti-M13 antibody. Rh-negative or
Rh-positive red blood cells were incubated with undiluted or with
five-fold serial dilutions (1/5, 1/25, 1/125, 1/625 and 1/3125) of
anti-Rh phage antibodies. The cards were centrifuged and samples
were assessed for agglutination. The modified Micro Typing System
card assay was capable of detecting anti-Rh agglutination at a
dilution of between 1/625 and 1/3125.
[0166] Procedures for Isolation of Tumor-Specific Antibodies
[0167] Fab/phage specific for tumor cells are useful for in vitro
diagnosis (lab assays of biopsy, fluid, or blood samples), in vivo
labeling of tumor/metastasis (coupling of antibody to imaging
probe), or for treatment of malignancy (coupling of antibodies to
chemical or radioactive toxins). Tumor-specific antibodies are also
useful for the identification of novel antigens or markers on tumor
cells which may form the basis for anti-tumor vaccines. Further,
tumor-specific antibodies useful for the generation of
anti-idiotypic antibodies may also form the basis for anti-tumor
vaccines.
[0168] Anti-tumor antibodies are generated essentially as described
herein for the generation of anti-Rh antibodies. Tumor cells, for
example, but not limited to, malignant melanoma cells, are
cell-surface biotinylated, labeled with streptavidin-magnetic
microbeads, and are then mixed with excess normal melanocytes.
Fab/phage libraries are generated from peripheral blood lymphocytes
of melanoma patients who possess therapeutically useful anti-tumor
antibodies. A number of melanoma patients who have "cured"
themselves apparently have done so by mounting a humoral (i.e.,
antibody) immune response. These Fab/phage libraries are incubated
with the admixture of cells. Fab/phage which are directed against
epitopes specific for malignant cells will bind to the malignant
cells and may then be isolated utilizing the magnetic column
panning approach.
[0169] Isolation of Fab/Phage that Identify Bacterial Virulence
Factors
[0170] The approach described herein may be used to isolate
Fab/phage capable of detecting differences between the virulent
bacteria and their nonpathogenic counterparts. In this case, the
virulent strain of bacteria is magnetically labeled, diluted with
the non-pathogenic counterpart, and an Fab/phage library which is
generated from lymphocytes obtained from individuals infected with
the virulent strain is added.
[0171] Fab/phage which are isolated in this manner may be useful
for the identification of novel bacterial antigens against which
antibacterial compounds and/or vaccines may be developed.
EXAMPLE 2
Genetic and Immunological Properties of Phage-Displayed Human
Anti-Rh(D) Antibodies
[0172] Clinically, the human Rh(D) antigen is the most important
red blood cell (RBC) membrane protein in transfusion medicine. The
autoimmune response against Rh(O) produces high affinity IgG
antibodies which cause hemolytic transfusion reactions and
hemolytic disease of the newborn (HDN). The prophylactic use of
Rh(D)-immune globulin in pregnant Rh(D)-negative women has been a
major advance in the prevention of HDN, yet the mechanism by which
the drug exerts its immune modulatory effect is not well
understood.
[0173] Monoclonal antibodies derived from the B cells of
Rh(D)-immune globulin donors have defined several dozen Rh(D)
epitopes (Scott, 1996, Transfus. Clin. Biol. 3:333). Paradoxically,
the Rh(D) antigen, a circa 30 kD transmembrane. protein, has
minimal extracellular mass and presents a very limited surface area
for epitope expression. Because molecular cloning of a large
repertoire of anti-Rh(D) antibodies has not previously been
performed, these observations remain non-reconciled.
[0174] Rational development of recombinant formulations of
Rh(D)-immune globulin would be facilitated by molecular cloning of
a large number of anti-Rh(D) antibodies. Such cloning would also
aid in the design of therapeutic agents that block antibody
binding. Furthermore, comprehensive genetic analysis of anti-Rh(D)
antibodies within a given alloimmunized individual would serve as a
paradigm for human immune repertoire development, an area of which
limited information is currently available. Previously, no more
than 8 IgG anti-Rh(D) human monoclonal antibodies have been derived
from a single individual (Boucher et al., 1997, Blood 89:3277).
[0175] In Example 1, a technique useful for isolating Fab/phage
antibodies directed against antigens expressed on cell surfaces was
described. Using this technique and intact human red blood cells
(RBCs), highly diverse .gamma..sub.1.kappa. and
.gamma..sub.1.lamda.. Fab/phage libraries against the Rh(D) antigen
from the B cells of a single Rh(D)-immune globulin donor were
generated.
[0176] In this Example, a detailed genetic and serological analysis
of 53 unique anti-Rh(D) antibodies derived from 83 randomly chosen
clones is presented. These data demonstrate extensive genetic
homology between antibodies directed against different Rh(D)
epitopes. Evidence is provided herein that antibodies directed
against different epitopes can be clonally related. Finally, a
model is described which reconciles the serological diversity of
anti-Rh(D) antibodies with the topological constraints imposed by
the Rh(D) antigen.
[0177] The materials and methods used in the experiments presented
in this Example are now described.
Production of Monoclonal Anti-Rh(p) Phage-Displayed and Soluble Fab
Molecules
[0178] Methods for the isolation of human anti-Rh(D)-specific
antibodies from .gamma..sub.1.kappa. and .gamma..sub.1.kappa.
Fab/phage display libraries using the pComb3H phagemid vector and a
cell-surface panning protocol have been described (Siegel et al.,
1997, J. Immunol. Meth. 206:73). Soluble anti-Rh(D) Fab
preparations for inhibition studies were produced from bacterial
cultures transfected with plasmid DNA from which the M13 gene III
coat protein sequence had been excised as described (Siegel et al.,
1994, Blood 83:2334; Barbas et al., 1991, Methods: A Companion to
Meth. Enzymol. 2:119). Cultures were grown by shaking at 300 RPM at
37.degree. C. in superbroth (30 g/L tryptone, 20 g/L yeast, 10 g/L
MOPS, pH 7.00) containing 20 mM MgCl.sub.2 and 50 mg/ml
carbenicillin to an OD.sub.600 of 0.5.
Isopropyl-.beta.-D-thiogalactopyranoside (IPTG) was added to 1 mM
and cultures were shaken overnight at 30.degree. C. Bacterial
pellets were harvested and resuspended in 1/50th of the initial
culture volume with osmotic shock buffer (500 mM sucrose, 1 mM
EDTA, 100 mM Tris, pH 8.00), incubated for 30' at 4.degree. C., and
centrifuged at 16,000.times. g for 15' at 4.degree. C.
Fab-containing supernatants were dialyzed against PBS and used in
agglutination experiments without further purification.
Anti-Rh(D) Antibody Binding Assays
[0179] The binding of anti-Rh(D) Fab/phage or soluble Fab molecules
to normal or partial Rh(D) antigens was assessed by indirect
agglutination assays as described (Siegel et al., 1994, Blood
83:2334; Siegel et al., 1997, J. Immunol. Meth. 206:73). Briefly,
100-.mu.l aliquots of phage-displayed Fabs or soluble Fabs were
incubated with 50 .mu.l of a 3% suspension of RBCs. Following a
one-hour incubation at 37.degree. C., the RBCs were washed 3 times
with 2 ml of cold PBS to remove unbound antibody. The resulting RBC
pellets were resuspended in 100 .mu.l of a 10 .mu.g/ml solution of
sheep anti-M13 antibody (5 Prime-3 Prime, Boulder, Colo.) for
Fab/phage experiments or goat anti-human .kappa. or .lamda. light
chain antibody (Tago, Burlingame, Calif.) for .gamma..sub.1.kappa.
or .gamma..sub.1.lamda. soluble Fab experiments, respectively. The
RBC suspensions were transferred to the round-bottomed wells of a
96-well microplate and left undisturbed for 5 2 hours. Negative
reactions show sharp .about.2 millimeter diameter RBC spots whereas
the RBCs in agglutinated wells form a thin carpet coating the
entire floor of the well (Siegel et al., 1997, J. Immunol. Meth.
206:73). Agglutination titers for recombinant antibodies were
determined by performing serial 2-fold dilutions in 1% BSA/PBS.
[0180] Typically, Fab/phage had agglutination titers of 1/1024 to
1/2048 (where "neat" is defined as 5.times.10.sup.12 cfu/ml; Siegel
et al., 1997, J. Immunol. Meth. 206:73) and soluble Fabs had
agglutination titers of 1/64 to 1/128 when prepared as described
above.
[0181] For determining Rh(D) epitope specificity for anti-Rh(D)
Fab/phage antibodies, the following reference Rh(D) variant cells
were used: O/D.sup.IIIaCce, G positive; B/D.sup.IIIcCce;
A/D.sup.IVace; A/D.sup.IVace; O/D.sup.IVaCce; O/D.sup.IVbCce;
B/D.sup.IVbCce, Go.sup.a negative, Rh32 negative; O/D.sup.VaCce;
O/D.sup.VacEe, D.sup.w positive; O/D.sup.VICce; B/D.sup.VICce;
AB/D.sup.VICce; A/D.sup.VIcEe; O/D.sup.VIICce; and O/D.sup.VIICce.
Each Fab/phage antibody was tested on at least 3 separate occasions
against at least 2 different examples of each variant cell type and
identical epitope assignments were obtained each time. For
antibodies that demonstrated not-previously-described patterns of
reactivity or repeatedly weak reactivity against one type of cell,
monoclonal Fab/phage were prepared on a least 4 separate occasions
to verify the patterns of reactivity.
[0182] For inhibition studies, the ability of antibodies with
different Rh(D) epitope specificities to compete with each other
for binding was assessed by preparing stocks of each clone in both
a soluble Fab form and a phage-displayed form. Pair-wise
combinations of soluble Fabs and Fab/phage were prepared and added
to Rh(D)-positive RBCs. The resulting incubation mixes comprised 50
.mu.l of a 3% suspension of RBCs, 100 .mu.l of undiluted soluble
Fab, and 100 .mu.l of Fab/phage diluted to its highest
agglutinating titer. Following a l-hour incubation at 37.degree.
C., RBCs were washed, resuspended in anti-M13 antibody, and placed
in microplate wells as described above. That the amount of soluble
Fab present in an incubation mixture was sufficient to compete away
a Fab/phage that shared the same binding site was determined by
verifying that each soluble Fab preparation could block its own
Fab/phage.
[0183] Inhibition experiments were also performed using pair-wise
combinations of soluble Fabs instead of soluble Fab and Fab/phage
combinations. In this type of experiment, pairs of soluble Fabs
specific for different epitopes were chosen such that one Fab
contained a .lamda. light chain and the other a .kappa. light
chain.
[0184] Incubations with RBCs were performed with one Fab in excess
and the other in limiting amounts. Blocking of the latter antibody
was assessed using a secondary antibody (anti-.lamda. or
anti-.kappa.) specific for its light chain isotype.
Nucleotide Sequencing and Analysis
[0185] Plasmid DNA for sequencing was prepared using the
Qiawell.TM. system (Qiagen, Chatsworth Calif.). Double-stranded DNA
was sequenced using light chain or heavy chain immunoglobulin
constant region reverse primers or a set of unique is pComb3H
vector primers that anneal 5' to the respective immunoglobulin
chain (Barbas et al., 1991, Methods: A Companion to Meth. Enzymol.
2:119; Roben et al., 1995, J. Immunol. 154:6437) and automated
fluorescence sequencing (Applied Biosystems, Foster City, Calif.).
Sequence analysis and variable region germline assignments were
performed using DNAplot (Althaus et al., 1996, DNAPLOT,
http://www.mrc_cpe.cam.ac.uk/imt_doc/DNAsearch.html) and the V Base
Directory of Human V Gene Sequences (March 97 update; Tomlinson et
al., 1996, V Base Directory of Human V Gene Sequences,
http://www.mrc_cpe.cam.ac.uk/umt_doc/vbase_home_page.htrl).
Germline assignments were corroborated with the MacVector (v. 6.0)
software package (Oxford Molecular Group, Oxford, UK) against the
same database. Multiple sequence alignments and predictions of
isoelectric point were calculated using the Pileup and Isoelectric
programs of the GCG software package (v. 8.0.1; GCG, Madison Wis.).
Statistical analysis was performed with Statview (Abacus Concepts,
Berkeley Calif.).
[0186] The results of the experiments presented in this Example are
now described.
[0187] Sequence Analysis of Anti-Rh(D) Heavy and Light Chains
[0188] Example 1 describes the use of Fab/phage display and
cell-surface panning to isolate a large array of anti-Rh(D)
antibodies from the peripheral blood lymphocytes of a single
hyperimmunized donor. Separate .gamma..sub.1.kappa. and
.gamma..sub.1.lamda. Fab/phage display libraries were constructed
and contained 7.times.10.sup.7 and 3.times.10.sup.8 independent
transformants, respectively, based on electroporation efficiency.
Each library was panned independently using a simultaneous
positive/negative selection strategy with magnetically-labeled
Rh(D)-positive RBCs and unmodified Rh(D)-negative RBCs as
described. Following two rounds of panning, 32 of 36
.gamma..sub.1.lamda. and 15 of 15 .gamma..sub.1.kappa. clones were
positive for anti-Rh(D) activity. After the third round of panning,
24 out of 24 .gamma..sub.1.lamda. and 12 out of 12
.gamma..sub.1.kappa. clones were positive. Nucleotide sequencing of
the 83 positive clones revealed a total of 28 unique heavy and 41
unique light chains. Due to combinatorial effects during phage
display library construction, heavy and light chain gene segments
paired to produce 53 unique Fab antibodies.
Anti-Rh(D) Heavy Chains
[0189] All of the heavy chain sequences used V.sub.HIII
family-encoded gene products, as indicated in FIGS. 7 and 8.
Several heavy chain sequences shared identical VDJ joining regions,
and 12 unique VDJ rearrangements were identified. These
rearrangements were designated VDJ1 through VDJ12. Alignment of
these sequences against the V Base Directory of Human V Gene
Sequences revealed that only four V.sub.HIII genes were used by
these antibodies: VH3-21, VH 3-30, VH 3-33, and VH 3-30.3. VH3-21
was used by 1 of the 12 VDJs and 2 of the 28 clones; VH3-30 was
used by 1 VDJ and 6 clones; VH3-33 was used by 9 VDJs and 19
clones; and VH3-30.3 was used by 1 VDJ and 1 clone. Interestingly,
VH3-30, VH3-33, and VH3-30.3 comprise a set of closely related
genes (>98% homology; FIG. 8B) and their next nearest neighbor,
VH3-07, is only 90% homologous (FIG. 8C). Hereafter, these three
genes are referred to as the "VH3-33 superspecies". Heavy chain E1
differed from VH3-21 by six mutations and differed from VH3-48 by
ten mutations; hence, it was assigned to the former germline gene.
Because there were no common mutations among the VH3-33 clones, it
is highly probable that the donor possessed the VH3-33 germline
gene. However, we could not formally rule out gene duplication with
allelic variants of VH3-33 or the existence of variant alleles of
the other germline genes in the donor. The isolation of clones
sharing multiple VDJ joining regions strongly suggests that cloning
artifacts cannot account for the V.sub.H restrictions observed.
[0190] Neither Jnor D segments showed restriction. At least 9
different D segments were used and J.sub.H gene utilization
comprised J.sub.H6 (5 VDJs and 9 clones), J.sub.H4 (4 VDJs and 10
clones), J.sub.H3 (2 VDJs and 8 clones) and J.sub.H.sup.5 (1 VDJ
and 1 clone). All four V.sub.H genes were Chothia class 1-3
(Chothia et al., 1992, J. Mol. Biol. 227:799), and the CDR3s showed
a narrow range of length from 15 to 19 residues.
[0191] Because rearranged heavy chain genes demonstrate extensive
diversity, clones sharing identical VDJ rearrangements are
generally considered to have arisen from the same clone. Based upon
nucleotide alignment with the germline genes, the ontogeny tree in
FIG. 9 was constructed for the 12 VDJs and 28 clones. By using the
most parsimonious mutation scheme (i.e. postulating the minimum
number of mutations), putative intermediate antibodies were derived
for several of the VDJs and were designated Ca, Cb, Da, Db, and Dc
(FIGS. 8A and 9). Compared with the isolated heavy chain clones,
which had between 6 and 23 nucleotide differences from their
germline counterparts, these putative intermediates had between 3
and 12 mutations from germline. Based upon the ontogeny tree, the
number of independent mutations could be tabulated among the
clones. The most commonly mutated residues were 52a and 58 (7
independent mutations), followed by residues 30, 31 and 50 (6 25
mutations), and residue 55 (5 mutations). In the VH3-33
superspecies, residues 52a and 58 in CDR2 are tyrosine residues and
residue 52a was mutated to phenylalanine in 6 of the 11 VDJs
derived from VH3-33 superspecies V.sub.H genes. Mutations at
residue 58 comprised glutamate (3), aspartate (2), histidine (1)
and asparagine (1). The AGY serines at residues 30, 31 and 55 were
mutated to a number of different amino acids, although the AGY
serine at 82b was conserved in all clones. The valine at residue 50
in the VH3-33 superspecies also had a diverse set of mutations.
This distribution of "hot spots" is similar to that seen with
non-productive rearrangements as previously reported by Dorner et
al (1997, J. Immunol. 158:2779).
Anti-Rh(D) Light Chains
[0192] Seventeen of the 18 .kappa., light chains were from the
V.sub..kappa.I family and the remaining light chain originated from
a V.sub..kappa.II family member germline gene (FIG. 10). Only four
V.sub..kappa. germline genes were used (15 clones were derived from
DPK9 alone), and the .kappa. light chain clones had between 1 and
49 mutations from their corresponding V.sub..kappa. germline genes.
All five of the known J.sub..kappa. genes were used and were each
joined to the DPK9 gene in one or more clones. Because the light
chains showed considerably less diversity in their joining regions
than the heavy chains, it was difficult to assign common clonal
origins. However, an ontogeny tree was constructed by grouping
common V and J gene segments along with common mutations. Based
upon this analysis, the 18 .kappa. chains comprised at least 10
different recombination events.
[0193] .lamda. light chains were restricted by their J.sub..lamda.
gene usage but showed no restriction in their use of V.sub..lamda.
genes (FIG. 11). The 23 .lamda. light chains all used the
J.sub..lamda.2Vasicek gene but were derived from V.sub..lamda.I (12
clones), V.sub..lamda.III (5), V.sub..lamda.VII (3),
V.sub..lamda.II (2) and V.sub..lamda.IV (1) family genes. The
number of mutations ranged from 2 to 41 from the nearest germline
V.sub..lamda. gene. Based upon common joining regions and
mutations, these 23 1 light chains were derived from at least 13
different B cells.
Assessment of the Diversity of the Non-Panned Libraries
[0194] In order to determine whether the apparent restriction in
gene usage of the anti-Rh(D) antibodies could have been due to
pre-selection factors (i.e. cloning artifacts), the diversity of
the non-panned .gamma..sub.1.kappa. and .gamma..sub.1.lamda.
Fab/phage libraries was assessed. By sequencing 39 randomly-picked
clones, we determined that there were no duplicate heavy or light
chain sequences, and that there was significant heterogeneity in V
gene family representation before selection (FIG. 12). In fact, the
variable region gene family distribution was not unlike that found
by other investigators for IgG-secreting lymphocytes in adult
peripheral blood (Stollar, 1995, Ann. NY Acad. Sci. 764:547).
Furthermore, of the 14 V.sub.HIII-encoded negative clones, only one
used a VH3-33 superspecies germline gene (VH3-30.3); the other 13
were encoded by VH3-07 (3), 3-09 (2), 3-15 (2), 3-48 (2), 3-72 (2),
3-23 (1), and DP-58 (1). Therefore, the restriction of the 83
anti-Rh(D) clones to the VH3-33, 3-30, 3-30.3 and 3-21 genes is
significant and not a result of skewed representation of certain
germline genes within the originally constructed
.gamma..sub.1.kappa. and .gamma..sub.1.lamda. Fab/phage
libraries.
Heavy and Light Chain Contribution to Rh(D) Epitope Specificity
[0195] Because of the conformational dependency of Rh(D)
antigenicity, Rh(D) "epitopes" have been classically defined
through the use of RBCs obtained from rare individuals whose cells
appear to produce Rh(D) antigens "lacking" certain epitopes.
Examining the pattern of agglutination of a particular anti-Rh(D)
monoclonal antibody with such sets of partial Rh(D) RBCs enables
one to categorize that antibody's fine specificity.
[0196] Monoclonal Fab/phage preparations were prepared in
triplicate for each of the 53 anti-Rh(D) clones and tested against
a panel of Rh(D) category cells IIIa/c, IVa, IVb, Va, VI, and VII.
This panel of cells can differentiate between the Rh(D) epitope
specificities as described by Lomas et al. (1989, Vox Sang 57:261;
designated epitopes epD1, epD2, epD3, epD4, epD5, and epD6/7).
Agglutination experiments using the Fab/phage clones demonstrated
five different patterns of reactivity, including a new pattern
which had not been described in the original study by Lomas et al.
or in the more recently-described (Scott, 1996, Transfus. Clin.
Biol. 3:333; Stollar, 1995, Ann. NY Acad. Sci. 764:547) 9-, 30-, or
37-epitope systems (as indicated by the data depicted in FIGS. 13
and 14). Although nearly all Fab/phage gave unequivocal
agglutination reactions, a few antibodies gave repeatedly weak
patterns of reactivity against one of the panel cells. For these
reactions, monoclonal Fab/phage were prepared on at least 4
separate occasions to verify the patterns of reactivity.
[0197] The most commonly-recognized epitope was epD6/7, against
which 13 of the clones described herein were directed.
Interestingly, monoclonal anti-Rh(D) clones isolated using
conventional tissue culture methods are most often specific for
epD6/7 (Mollison et al., 1993, In: Blood Transfusions in Clinical
Medicine 9th ed., Blackwell Scientific, Oxford, U.K.). EpD2, epD1,
and epD3 were recognized by 10, 7, and 2 clones, respectively. Six
clones agglutinated cells of categories ma/c, IVa, and VII, but not
of categories IVb, Va, and VI, and were designated anti-"epDX" .
This pattern is identical to epD1, except that the IVa cell is
agglutinated. Three clones gave intermediate reactions with cell
IVa, but otherwise showed patterns consistent with epDX or epD1.
These clones were designated epDX.sup.1 or epD1.sup.X depending on
whether this reactivity against cell IVa was stronger or weaker,
respectively (see FIG. 14). Similarly, reaction patterns for epD1
and epD2 differ by a positive reaction with the category Va cell;
therefore, one clone was given epD2.sup.1 specificity because it
gave only moderate reactivity against that cell. Such variable
reactions against one or more partial Rh(D) cells have been
observed for anti-Rh(D) monoclonal antibodies produced through
conventional tissue culture methods Tippett et al., 1996, Vox Sang.
70:123).
[0198] Because of the reassortment of heavy and light chain gene
segments that occurs during the construction of a phage display
library, a number of clones were isolated that shared either a
heavy (e.g. E1) or light (e.g. M3) chain sequence (FIG. 14). Some
heavy chains were found to have paired with both .kappa. and
.lamda. light chains (e.g. C1, D20) and each demonstrated
anti-Rh(D) specificity. Interestingly, some heavy chains (e.g. E1,
D12) mapped to different epitopes depending upon the light chains
with which they were paired. In particular, the light chains of two
such clones, E1/M2 and E1/M3, differed by only three amino acid
residues (FIG. 11) and these differences appear to confer
specificity for epD2 vs. epD3.
Inhibition Studies
[0199] To investigate the topological relationships among the Rh(D)
epitopes, inhibition studies were performed. Gorick et al. (1988,
Vox Sang. 55:165) used pairs of non-labeled and .sup.125I-labeled
anti-Rh(D) monoclonal antibodies to demonstrate that antibodies to
at least three different Rh(D) epitopes (subsequently identified as
epD1, D6 and D7; Lomas et al., 1989, Vox Sang. 57:261) could
inhibit one another. Recombinant antibodies to five Rh(D) epitopes
were used to confirm and extend those findings (FIG. 15). In one
series of experiments, the ability to express each antibody in both
a soluble Fab as well as phagelisplayed form was exploited to
determine whether a soluble Fab directed against one epitope would
inhibit the agglutination induced by an Fab/phage directed against
a different epitope. Reciprocal pairs of soluble Fab and Fab/phage
specific for epD1, epD2, epD3, epD6/7, and epDX were tested. All
ten combinations showed mutual inhibition patterns (illustrated in
FIG. 15A for an anti-epD3/anti-epD6/7 combination). To show that
this inhibition was not due to non-specific factors, a control with
an irrelevant RBC-binding recombinant antibody (an anti-blood group
B antibody) was performed (FIG. 1 SB). That sufficient inhibitory
amounts of soluble Fab was present were first verified by
demonstrating that each soluble Fab could inhibit its own Fab/phage
(FIGS. 15A and 15B; samples on diagonal). Similar results were
obtained using pairs of soluble Fabs which differed in their light
chain isotype composition (FIG. 15C).
Isoelectric Point Analysis of Anti-Rh(D) Antibodies
[0200] The restriction in V.sub.H germline gene usage to only four
V.sub.HIII family members was intriguing in light of their ability
to confer specificity to a number of Rh(D) epitopes. V.sub.H
germline gene segments used to encode anti-Rh(D) antibodies are
among the most cationic segments available in the human V.sub.H
repertoire which may be used to account for the relatively high pI
of polyclonal anti-Rh(D)-containing antisera (Boucher et al., 1997,
Blood 89:3277; Abelson et al., 1959, J. Immunol. 83:49; Frame et
al., 1969, Immunology 16:277). Although the cationic nature of the
antibodies may be important for binding to Rh(D), a constitutive
net positive charge may be necessary to permeate the highly
negative RBC .zeta. potential, thus permitting antibody to contact
antigen (Mollison et al., 1993, In: Blood Transfusion in Clinical
Medicine, 9th ed., Blackwell Scientific, Oxford, U.K). In either
case, analysis of the predicted pI for the 28 heavy chains and 41
light chains isolated here showed an interesting phenomenon for the
heavy chains, as compared with the light chains. Using the pI
interval scale of Boucher et al. (1997, Blood 89:3277), the average
pI of the four germline V.sub.H segments used to encode the 28
heavy chains is high (9.87.+-.0.15), significantly higher than that
of 39 randomly-picked, non-Rh(D) binding clones from the original
non-panned libraries (9.24.+-.0.80, P<10.sup.-5). Similar to the
results of Boucher et al., the addition of D and J.sub.H segments
and the introduction of somatic mutation did not significantly
change the pI of the average anti-Rh(D) heavy chain (9.81.+-.0.33,
P<0.37). However, for the light chains, the average pI of their
germline counterparts was not cationic, but the light chains became
so through the addition of J.sub.L segments and somatic mutation.
Overall, for all 18.kappa. and 23.lamda. light chains, paired
t-test analyses before and after somatic mutation showed a
significant increase in net positive charge when comparing germline
V.sub.L (6.63.+-.1.47) with expressed V.sub.L (7.28.+-.1.51 ,
P<10.sup.-3) or germline V.sub.LJ.sub.L (7.43.+-.1.47) with
expressed V.sub.LJ.sub.L (8.55.+-.1.35, P<10.sup.-7). There was
no significant increase in a similar analysis of 16 non-Rh(D)
binding clones (P<0.59 and P<0.19, respectively). Examination
of the light chain sequences listed in FIGS. 10 and 11 revealed
that this increase in pI resulted from mutations that not only
introduced positively-charged residues, but also eliminated some
negatively-charged residues. There were 31 such events, 29 (91%) of
which occurred in the light chain CDR regions.
Conventional and Phage-Displayed Anti-Rh(D) Monoclonal
Antibodies
[0201] The phage-display derived anti-Rh(D) clones were compared
with those produced by conventional tissue culture techniques
(EBV-transformation and cell fusion). Despite the relatively small
number of previously-published sequences for IgG anti-Rh(D)
antibodies (N=21) and the fact that they were derived from over 10
different donors, there was surprisingly good correlation between
the two groups, as indicated in Table 3. Both cohorts demonstrated
a predominance of V.sub.HIII-family encoded germline genes,
particularly those of the VH3-33 superspecies. CDR3 regions had
similar lengths ranging from 15-19 residues for Fab/phage
antibodies and 16-20 for conventional monoclonal antibodies,
although one heterohybridoma was an outlier, having a CDR3 length
of 28 residues. .kappa. light chains were biased towards
V.sub..kappa.I family members and .lamda. light chains demonstrated
the preferential use of the J.sub..lamda.2Vasicek gene. The only
qualitative discrepancy was in V.sub..lamda. family usage where
Fab/phage clones demonstrated a slight preference for
V.sub..lamda.I vs. V.sub..lamda.III family members for conventional
monoclonal antibodies. However, in both cohorts, DPL16 was used
more often than any other A light chain gene. TABLE-US-00007 TABLE
3 Comparison of IgG Fab/phage library-derived anti-Rh(D) monoclonal
antibodies prepared as described herein with those previously
produced by conventional tissue culture methods Previously Current
Study Attribute Published* (by clone).dagger. (by VDJ) Heavy Chains
VH3 family derived 12/21 (57%) 28/28 (100%) 12/12 (100%) VH3-33
10/12 (83%) 26/28 (93%) 11/12 (92%) superspecies.dagger-dbl./VH3
VH3-33/VH3 9/12 (75%) 19/28 (68%) 9/12 (75%) VH3-21/VH3 1/12 (8%)
2/28 (7%) 1/12 (8%) VH4-34 derived 2/21 (10%) 0/28 (0%) 0/12 (0%)
JH6 usage 15/21 (71%) 9/28 (32%) 5/12 (42%) CDR3 length 16-20
(28.sctn.) 15-19 .kappa. Light Chains V.kappa.1 family 8/12 (67%)
17/18 (94%) derived/total .kappa. J.kappa.1 usage/total .kappa.
4/12 (33%) 6/18 (33%) J.kappa.2 usage/total .kappa. 4/12 (33%) 6/18
(33%) .lamda. Light Chains V.lamda.1 family 2/8 (25%) 12/23 (52%)
derived/total .lamda. V.lamda.3 family 5/8 (63%) 5/23 (22%)
derived/total .lamda. DPL16 derived/ 3/5 (60%) 4/5 (80%) V13 family
J.lamda.2Vasicek 6/8 (75%) 23/23 (100%) usage/total .lamda. Notes
for Table 3 *Compiled from a total of 21 sequences of IgG
anti-Rh(D) antibodies isolated from multiple subjects originally
published by Bye et., Hughes -Jones et al., Chouchane et al., and
Boucher et al. and available from Genbank. One light chain (Oak-3)
was not available in Genbank and was not included in the
assessment. .dagger.For heavy chains, left column tabulates each
clone separately; right column tabulates clones on the basis of
shared V-D-J joining regions .dagger-dbl.VH3-33 superspecies
defined as the group of VH3 family germline genes comprising
VH3-33, VH3-30, and VH30.3. .sctn.CDR3 length outlier
[0202] It has been suggested in the literature that the VH4-34
(VH4.21) germline gene, a gene used by many autoantibodies and cold
agglutinins, may play an important role in the immune response to
Rh(D) (Silberstein et al., 1991, Blood 78:2377; Pascuel et al.,
1991, J. Immunol. 146:4385; Silverman et al., 1988, J. Exp. Med.
168:2361; Thompson et al., 1991, Scand. J. Immunol. 34:509).
However, these conclusions arose from the analysis of IgM
monoclonal antibodies and only 2 of the 21 published anti-Rh(D) IgG
sequences used VH4-34 (Bye et al., 1992, J. Clin. Invest. 90:2481).
In a related series of experiments, aliquots of the
.gamma..sub.1.kappa. and .gamma..sub.1.lamda. libraries obtained
after the second and third rounds of selection were pooled and then
panned against the VH4-34 specific rat anti-idiotypic monoclonal
antibody (9G4; Stevenson et al., 1989, Br. J. Haematol. 72:9).
Although VH4-34 encoded antibodies were successfully enriched, the
Fab/phage were not specific for Rh(D) and displayed serological
characteristics similar to those of cold agglutinins.
Rh(D) Epitones and Significance of Antibody Sequences
[0203] Since the initial report by Argall et al. in 1953 (J. Lab.
Clin. Med. 41:895), it has been recognized that rare individuals
who type as Rh(D)-positive can produce allo-anti-Rh(D) antibodies
in response to Rh(D) immunization by transfusion or pregnancy. This
phenomenon was explained by hypothesizing that the Rh(D) antigen is
a "mosaic structure" and that these individuals were producing
alloantibodies to parts of the mosaic they lack. By systematically
examining patterns of reactivity between their cells and sera, RBCs
expressing partial Rh(D) antigens were divided into categories,
each presumed to have a different abnormality in their Rh(D)
antigen.
[0204] Through the subsequent use of index panels of monoclonal
anti-Rh(D) antibodies, a series of epitopes were defined of which
the number and combination varied from one Rh(D) category to
another. As new monoclonal antibodies were produced, their
reactivity profiles against these partial Rh(D) RBCs became the
standard method for determining Rh(D) antibody epitope specificity.
Molecular analyses of partial Rh(D) phenotypes have shown that the
Rh(D) genes in these individuals have either undergone intergenic
recombination with the highly homologous Rh(CE) gene, or, less
commonly, have sustained point mutation(s) (Cartron et al., 1996,
Transfus. Clin. Biol. 3:497).
[0205] As noted earlier, to investigate the topological
relationships among Rh(D) epitopes, Gorick et al. performed
competition experiments with Rh(D) monoclonal antibodies and
observed varying degrees of inhibition (Gorick et al., 1988, Vox
Sang. 55:165). These results, when combined with those of Lomas et
al. (1989, Vox Sang. 57:261), suggested a model for Rh(D) in which
epitopes are spatially distinct yet demonstrate a certain degree of
overlap as illustrated in FIG. 16A. This model explained how
antibodies to two different Rh(D) epitopes (in this case epD2 and
epD3) could inhibit each other's binding to wild type Rh(D), and
how a change in the structure of Rh(D) in category VI RBCs
(asterisk in FIG. 16A) would cause the loss of epD2. However, based
upon this concept of Rh(D) epitopes as distinct domains, one would
expect that antibodies against different epitopes of Rh(D) would be
structurally and genetically distinct as well. Thus, it was
surprising that the anti-Rh(D) clones described herein demonstrated
such marked restriction in gene usage. For example, only two
superspecies of V.sub.H genes were used despite specificities for 4
of the original 6 Rh(D) epitopes described by Lomas et al. (1989,
Vox Sang. 57:261). Furthermore, multiple specificities could arise
from a single heavy chain depending upon the light chain with which
it was paired (e.g. El with M2, M3, L3, or L4). In addition, other
clones repeatedly demonstrated variable weak reactivity against
certain Rh(D) category RBCs that would affect the epitope
specificities to which they were assigned (e.g. C1 with O1, M1, or
J5).
[0206] Several hypotheses could account for these findings. The
most simplistic interpretation is that the heavy chain does not
directly interact with the antigen, but rather is responsible for
bringing the antibody in close proximity with the antigen. The
specific interactions between the light chain and the antigen would
then determine the epitope specificity for that antibody. In this
regard, the data presented herein are consistent with the
observations of Boucher et al. (1997, Blood 89:3277) on the
relative cationic nature of anti-Rh(D) heavy chains. However,
because it was determined during the studies described herein that
light chains become cationic during somatic mutation, the charge of
the entire antibody may play a role in its ability to bind,
resulting in the selection and expansion of particular B-cell
clones.
[0207] A more compelling hypothesis is that Rh(D) epitopes do not
differ spatially but differ only in the number and arrangement of
contact residues presented, as illustrated in FIG. 16B. In other
words, the "footprints" of most, if not all, anti-Rh(D) antibodies
are essentially identical to one another. The genetic events which
produce partial Rh(D) molecules result in the loss of certain
critical key points of contact necessary for some antibodies to
bind; alternatively, they result in the formation of new structures
that interfere with the binding of other anti-Rh(D)
immunoglobulins. For example, the introduction of a "ledge" in
Rh(D) category VI cells (asterisk in FIG. 166B) does not interfere
with the binding of an anti-epD3 antibody, but does prevent the
binding of anti-epD2. Therefore, category VI RBCs are said to have
epD3 but "lack" epD2.
[0208] This model is consistent with the inhibition experiments
described herein (e.g. FIG. 15) and with those of Gorick et al.
(1988, Vox Sang. 55:165) and offers an explanation for the marked
restriction in heavy chain gene usage. This model also reconciles a
mechanism by which one heavy chain (e.g. E1) can confer binding to
multiple epitopes and why some of the recombinant anti-Rh(D)
antibodies described herein, as well as some
conventionally-produced monoclonal antibodies (e.g. Tippett et al.,
1996, Vox Sang. 70:123), display variable reactivity against
certain categories of partial Rh(D) RBCs. From the antigen's
perspective, this model explains how a single point mutation in
Rh(D) can result in the loss of multiple Rh(D) epitopes (such as
T2831 in category HMi RBCs) and how the residues associated with
the expression of some epitopes appear to be distributed among
nearly all the extracellular loops of Rh(D). It also provides an
understanding as to how .gtoreq.37 "epitopes" can fit on the
relatively small extracellularly-exposed surface of the Rh(D)
molecule.
[0209] This concept of "coincident" epitopes is best exemplified by
comparing the E1/M2 and E1/M3 clones described herein. The only
difference between the reactivity of E1/M2 and E1/M3 is the ability
of the latter antibody to agglutinate Rh(D) category VI cells, as
depicted in FIG. 13. Hence, E1/M2 is classified as an anti-epD2 and
E1/M3 as an anti-epD3 antibody. Light chains M2 and M3 differ by
only 3 residues: D82A, G95aA, and W96V, as indicated in FIG. 11.
Therefore, some combination of these three residues confers
reactivity against category VI cells. In other words, epD2 and
epD3, as seen by the E1/M2 and E1/M3 antibodies, differ by the
binding constraints imposed by at most three mutations. If the
model depicted in FIG. 16A were correct and the epitopes were
independent, these mutations would have to cause enough structural
alteration in the antibody combining site so that a completely
separate epitope on the same antigen would be recognized. It would
seem unlikely that these 3 mutations could cause such a change,
especially given the lack of internal homology domains in Rh(D).
Therefore, it is concluded that it is far more plausible that the
footprints of these 2 antibodies are essentially identical, and
that one or more of these mutations (e.g. the tryptophan in CDR3 of
M2) prevent(s) the interaction of E1/M2 with category VI RBCs.
Since other clones demonstrate that the light chain can confer
specificity against epD1, epD2, or epD3 (with the E1 heavy chain);
epD1 or epDX (with CS); and epD1, epD2, and epD6/7 (with D12), we
suggest that all 5 of these epitopes have similar antibody
combining sites.
Immunologic and Clinical Implications of Proposed Model
[0210] The model depicted in FIG. 16B leads to additional
predictions concerning the Rh(D) immune response beyond simply
clarifying what is meant by an Rh(D) epitope. It is commonly stated
in the transfusion medicine literature that individuals whose RBCs
express partial Rh(D) antigens are free to make antibodies to the
Rh(D) epitopes they lack (Mollison et al., 1993, In: Blood
Transfusion in Clinical Medicine, 9th ed. Blackwell Scientific,
Oxford, U.K.). Therefore, an individual who produces category VI
RBCs should be able to make anti-epD2 but not anti-epD3. If these
epitopes were truly independent, then the immune repertoire of the
anti-epD2 antibodies made by a category VI individual would be
similar to those produced by an Rh(D)-negative person. However, to
the immune system, epD2 and epD3 are not independent.
[0211] It is herein postulated that somatic mutation of an
anti-epD3 antibody can change its fine specificity to that of epD2
(or vice-versa, see FIG. 16C). Suppose that the preferred way of
making an anti-epD2 antibody is through an anti-epD3 intermediate.
To an Rh(D)-negative individual, this process can take place
unimpeded. However, in a category VI individual, this route would
be unfavorable because an anti-epD3 antibody would be
self-reactive. As a result, such an individual would have to make
anti-epD2 antibodies by alternative routes or by tolerating some
degree of auto-reactivity in the process. With respect to the
latter point, it is of interest to note that a transient production
of auto-anti-Rh(D) frequently precedes or accompanies the early
production of allo-anti-Rh(D) in individuals who express partial
Rh(D) antigens (Chown et al., 1963, Vox Sang. 8:420; Macpherson et
al., 1966, J. Clin. Pathol. 45:748; Beard et al., 1971, Med. Genet.
8:317; Cook 1971, Br. J. Haematol. 20:369; Holland et al.,
Transfusion 13:363 (Abstract); Issit, 1985, In: Applied Blood Group
Serology 3rd ed., Montgomery Scientific, Miami Fla.). It is
predicted, therefore, that the anti-epD2 antibodies from a category
VI individual would be different in composition (i.e. gene usage)
and quite possibly quantitatively depressed as compared to an
Rh(D)-negative individual. This may be analogous to the antibodies
of the ABO blood group system in which it has been observed that
anti-A and anti-B titers in blood group O individuals are
significantly higher than in blood group B or A individuals,
respectively (Ichikawa, 1959, Jap. J. Med. Sci. Biol. 12:1). Blood
group O individuals are unconstained in creating their anti-A and
anti-B immune repertoires while individuals who produce A s or B
antigens (2 nearly identical structures) must do so in a manner
that avoids self-reactivity.
[0212] In the case of antibodies E1/M2 and E1/M3, they appear to
have arisen from a common precursor B cell rather than directly
from each other (FIG. 11). To test the framework of the hypothesis
presented herein, i.e. somatic mutation resulting in "epitope
migration" of an antibody, one may construct the precursors and
potential intermediates between the M2 and M3 light chains and then
determine what Rh(D) epitope specificities (if any) they express.
This concept of epitope migration has been previously reported for
murine anti-cryptococcal and anti-type II collagen antibodies
(Mukheijee et al., 1995, J. Exp. Med. 181:405; Mo et al., 1996, J.
Immunol. 157:2440).
[0213] If the model proposed herein for Rh(D) epitopes is correct,
then the question of the number of epitopes may be obsolete. There
may be as many epitopes as can be differentiated by the number of
cell categories, i.e. 2.sup.n epitopes where n is the number of
distinct partial Rh(D) RBCs.
[0214] A more important question is the interrelationships between
the various epitopes. For example, are some epitopes "further away"
than others--not in the topological sense, but in terms of the
number of mutational hits an antibody needs to receive in order to
change its serologic reactivity. Furthermore, does the humoral
immune response in a partial Rh(D) individual differ from that in
an Rh(D)-negative individual in the manner predicted by this model?
One may find that allo-anti-Rh(D) antibodies made by partial Rh(D)
individuals are not as clinically significant, i.e. capable of
inducing hemolysis. This may explain why hemolytic disease of the
newborn due to anti-Rh(D) produced by pregnant individuals with
partial Rh(D) phenotypes is so rare even when taking into account
the low prevalence of the partial Rh(D) phenotypes (Mollison et
al., 1993, In: Blood Transfusion in Clinical Medicine, 9th ed.
Blackwell Scientific, Oxford, U.K). A better understanding of the
immune response to Rh(D) in these patients may alleviate concerns
regarding the need to identify such individuals to ensure that they
only receive Rh(D)-negative blood products for transfusion and
Rh(D)-immune globulin during pregnancy (Jones et al., 1995, Trans.
Med. 5:171). Furthermore, with respect to the design of recombinant
Rh(D)-immune globulin for use in Rh(D)-negative patients, it may
not be necessary to formulate cocktails of monoclonal antibodies
containing multiple Rh(D) epitope specificities.
Sequence Data
[0215] Genbank accession numbers for anti-Rh(D) heavy chains are as
follows: B01, AF044419; C01, AF044420; C03, AF044421; C04,
AF044422; C05, AF044423; C08, AF044424; C10, AF044425; D01,
AF044426; D03, AF044427; D04, AF044428; D05, AF044429; D07,
AF044430; D08, AF044431; D09, AF044432; D10, AF044433; D11,
AF044434; D12, AF044435; D13, AF044436; D14, AF044437; D15,
AF044438; D16, AF044439; D17, AF044440; D18, AF044441; D20,
AF044442; D30, AF044443; D31, AF044444; E01, AF044445; E03,
AF044446.
[0216] Genbank accession numbers for anti-Rh(D) .kappa. light
chains are as follows: F01, AF044447; G01, AF044448; H01, AF044449;
I01, AF044450; I02, AF044451; I03, AF044452; I04, AF044453; I05,
AF044454; I06, AF044455; I07, AF044456; I08, AF044457; I09,
AF044458; I10, AF044459; I11, AF044460; I12, AF044461; I13,
AF044462; I15, AF044463; I16, AF044464.
[0217] Genbank accession numbers for anti-Rh(D) .lamda. light
chains are as follows: J01, AF044465; J02, AF044466; J04, AF044467;
J05, AF044468; K01, AF044469; K02, AF044470; K03, AF044471; L01,
AF044472; L03, AF044473; L04, AF044474; L05, AF044475; M01,
AF044476; M02, AF044477; M03, AF044478; N01, AF044479; N02,
AF044480; O01, AF044481; O02, AF044482; O03, AF044483; P01,
AF044484; Q01, AF044485; R01, AF044486; S01, AF044487.
Amino Acid Sequences of Anti-Rh(D) Heavy and Light Chains
[0218] The amino acid sequences of various anti-Rh(D) chains are
represented using single letter amino acid codes, as described
herein.
[0219] The amino acid sequence of the anti-Rh(D) chain B01 is
TABLE-US-00008 EVQLLESGGGVVQPGRSLRLSCAASGFTFRSYAMHWV (SEQ ID NO:1)
RQAPGKGLEWVAATAYDGKNKYYADSVKGRFTISRDN
SKNTLFLQMNSLRAEDTAVFYCARGGFYYDSSGYYGL RHYFDSWGQGTLVTVSS.
[0220] The amino acid sequence of the anti-Rh(D) chain C01 is
TABLE-US-00009 EVQLLESGGGVVQPGRSLRLSCAASGFSFSSYGMHWV (SEQ ID NO:2)
RQAPGKGLEWVSVISYDGHHKNYADSVKGRFTISRDN
SKKTLYLQMNSLRPEDTAVYYCANLRGEVTRRASVPF DIWGPGTMVTVSS.
[0221] The amino acid sequence of the anti-Rh(D) chain C03 is
TABLE-US-00010 EVQLLESGGGVVQHGRSLRLSCAASGFSFSSYGMHWV (SEQ ID NO:3)
RQAPGKGLEWVSVISYDGHHKNYADSVKGRFTISRDN
SKKTLYLQMNSLRPEDTAVYYCANLRGEVTRRASVPF DIWGPGTMVTVSS.
[0222] The amino acid sequence of the anti-Rh(D) chain C04 is
TABLE-US-00011 (SEQ ID NO: 4) EVQLLESGGGVVQPGRSLRLSCAASGFSFSTYGMHWV
RQAPGKGLEWVSVISYDGHNKNYADSVKGRFTISRDN
SKKTLYLQMNSLRPEDTAVYYCANLRGEVTRRASIPF DIWGQGTMVTVSS.
[0223] The amino acid sequence of the anti-Rh(D) chain C05 is
TABLE-US-00012 (SEQ ID NO: 5) EVQLLESGGGVVQPGRSLRLSCAASGFSFSSYGMHWV
RQAPGKGLEWVAVISYDGTNKYFADSVKGRFTISRDN
SKKTLYLQMTSLRPEDTAVYFCANLRGEVTRRASVPL DIWGQGTMVTVSS.
[0224] The amino acid sequence of the anti-Rh(D) chain C08 is
TABLE-US-00013 (SEQ ID NO: 6) EVQLLESGGGVVQPGRSLRLSCAASGFSFSSYGMHWV
RQAPGKGLEWVAVISYDGTNKYFADSVKGRFTISRDN
SKKTLYLQMTSLRPEDTAVYFCANLRGEVTRRASVPL DIWGQGTMVTVSS.
[0225] The amino acid sequence of the anti-Rh(D) chain CIO is
TABLE-US-00014 (SEQ ID NO: 7) EVQLLESGGGVVQPGRSLRLSCAASGFSFSSYGMHWV
RQAPGKGLEWVSVISYDGHHKNYADSVKGRFTISRDN
SKKTLYLQMNSLRPEDTAVYYCANLRGEVTRRASVPF DIWGPGTLVTVSS.
[0226] The amino acid sequence of the anti-Rh(D) chain D01 is
TABLE-US-00015 (SEQ ID NO: 8) EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHWV
RQAPGKGLEWVAVIWFDGSNKYYADSVKGRFTISRDN
SKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRYLYY FDYWGQGTLVTSS.
[0227] The amino acid sequence of the anti-Rh(D) chain D03 is
TABLE-US-00016 (SEQ ID NO: 9) EVQLLESGGGVVQPGRSLRLSCAASGFTFSTYGMHWV
RQAPGKGLEWVAVIWFDGSNKEYADSVKGRFTVSRDN
SKNTLYLQMNSLRAEDTAVYYCAREEVVRGVILWSRK FDYWGQGTLVTVSS.
[0228] The amino acid sequence of the anti-Rh(D) chain D04 is
TABLE-US-00017 EVQLLESGGGVAQPGRSLRLSCVASGFSLRSYGMHW (SEQ ID NO: 10)
VRQAPGKGLEWVADIWFDGSNKDYADSVKGRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCARDWRVRAFSSG WLSAFDIWGQGTTVSVSS.
[0229] The amino acid sequence of the anti-Rh(D) chain D05 is
TABLE-US-00018 EVQLLEESGGGVAQPGRSLRLSCVASGFSLRSYGMH (SEQ ID NO: 11)
WVRQAPGKGLEWVADIWFDGSNKDYADSVKGRFTIS
RDNSKNTLYLQMNSLRAEDTAVYYCARDWRVRAFSS GWLSAFDIWGQGTTVSVSS.
[0230] The amino acid sequence of the anti-Rh(D) chain D07 is
TABLE-US-00019 EVQLLESGGGVVQPGRSLRLSCAVSGFTLTNYGMHW (SEQ ID NO: 12)
VRQAPGKGLEWVAHVWYDGSKTEYADSVKGRFAVSR
DKSKNTLFLQMNSLTAEDTAIYYCARERREKVYILF YSWLDRWGQGTLVTVSS.
[0231] The amino acid sequence of the anti-Rh(D) chain D08 is
TABLE-US-00020 EVQLLEESGGGVVQPGRSLRLSCAASGTFTSSYGMH (SEQ ID NO: 13)
WVRQAPGRGLEWVALIWYDGGNKEYADSVKGRFSIS
RDNSKNTLYLQVNSLRADDTAVYYCARDQRAAAGIF YYSRMDVWGQGTTVTVSS.
[0232] The amino acid sequence of the anti-Rh(D) chain D09 is
TABLE-US-00021 EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGMHW (SEQ ID NO: 14)
VRQAPGKGLEWVAFIWFDGSNKYYEDSVKGRFTVSR
DNSKNTLYLQMNSLRAEDTAVYYCAREGSKDVALSR YYYYMDVWGQGTTVTVSS.
[0233] The amino acid sequence of the anti-Rh(D) chain D10 is
TABLE-US-00022 EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGMHW (SEQ ID NO: 15)
VRQAPGKGLEWVAFIWFDGSNKYYEDSVKGRFTVSR
DNSKNTLYLQMNSLRAEDTAVYYCAREVSKKVALSR YYYYMDVWGQGTTVTVSS.
[0234] The amino acid sequence of the anti-Rh(D) chain D11 is
TABLE-US-00023 EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGMHW (SEQ ID NO: 16)
VRQAPGEGLEWVAFIWFDGSNKYYADSVKGRFTVSR
DNSKNTLYLQMNSLRAEDTAVYYCAREVSKKLALSR YYYYMDVWGQGTTVTVSS.
[0235] The amino acid sequence of the anti-Rh(D) chain D12 is
TABLE-US-00024 EVQLLESGGGVVQPGRSLRLACAASGFSFRSYGMHW (SEQ ID NO: 17)
VRQAPGRGLEWVAFTWFDGSNKYYVDSVKGRFTISR
DNSKNTLYLEMNSLRVDDTAVYYCAREASMLRGISR YYYAMDVWGPGTTVTVSS.
[0236] The amino acid sequence of the anti-Rh(D) chain D13 is
TABLE-US-00025 EVQLLESGGGVVQPGRSLRLSCAASGFTFSTYGMHW (SEQ ID NO: 18)
VRQAPGKGLEWVAVIWFDGSNRDYAESVKGRFTISR
DKSKNTLYLQMNSLRAEDSAVYYCARENVARGGGGV RYKYYFDYWGQGTLVTVSS.
[0237] The amino acid sequence of the anti-Rh(D) chain D14 is
TABLE-US-00026 EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYGMHW (SEQ ID NO: 19)
VRQAPGKGLEWVAVIWFDGSKRDYAESVKGRFTISR
DNSKNTLYLQMNSLRAEDSAVYYCARENVARGGGGI RYKYYFDYWGQGTLVTVSS.
[0238] The amino acid sequence of the anti-Rh(D) chain D15 is
TABLE-US-00027 EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHW (SEQ ID NO: 20)
VRQAPGKGLEWVAVIWFDGSNKYYADSVKGRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRY LYYFDYWGQGTLVTVSS.
[0239] The amino acid sequence of the anti-Rh(D) chain D16 is
TABLE-US-00028 EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHW (SEQ ID NO: 21)
VRQAPGKGLEWVAVIWFDGSNKYYADSVKGRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRY LYYFDYWGQGTLVTVSS.
[0240] The amino acid sequence of the anti-Rh(D) chain D17 is
TABLE-US-00029 EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHW (SEQ ID NO: 22)
VRQAPGKGLEWVAVIWFDGSNKYYADSVKGRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRY LYYFDYWGQGTLVTVSS.
[0241] The amino acid sequence of the anti-Rh(D) chain D18 is
TABLE-US-00030 EVQLLESGGGVVQPGRSLRLSCVVSGFTFNNYGMHW (SEQ ID NO: 23)
VRQASGKGLEWVAVIWFDGSNKYYADSVKGRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCARENQIKLWSRY LYYFDYWGQGTLVTVSS.
[0242] The amino acid sequence of the anti-Rh(D) chain D20 is
TABLE-US-00031 EVQLLESGGGVVQPGRSLRLSCAASGFTFSTYGMHW (SEQ ID NO: 24)
VRQAPGKGLEWVAVIWFDGSNKEYADSVKGRFTISR
DNSKNTLYLQMNSLRAEDTAVYYCAREEVVRGVILW SRKFDYWGQGTLVTVSS.
[0243] The amino acid sequence of the anti-Rh(D) chain D30 is
TABLE-US-00032 EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMRW (SEQ ID NO: 25)
VRQAPGKGLEWVAVVYYDGSNKHYSDSVKGRFTISR
DNSKNTLYLQMDSLRAEDTAVYYCARERNFRSGYSR YYYGMDVWGPGTTVTVSS.
[0244] The amino acid sequence of the anti-Rh(D) chain D31 is
TABLE-US-00033 EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGMHW (SEQ ID NO: 26)
VRQAPGKGLEWVAVVYYDGSNKHYSDSVKGRFTISR
DNSKNTLYLQMDSLRAEDTAVYYCARERNFRSGYSR YYYGMDVWGPGTTVTVSS.
[0245] The amino acid sequence of the anti-Rh(D) chain E01 is
TABLE-US-00034 EVQLLESGGGLVKPGGSLRLSCAASGFTFSSYSMHW (SEQ ID NO: 27)
VRQAPGKGLEWVSSISNSNTYIYYADAVKGRFTISR
DNAKNSLYLQMNSLRAEDTAVYYCARDSRYSNFLRW VRSDGMDVWGQGTTVIVSS.
[0246] The amino acid sequence of the anti-Rh(D) chain E03 is
TABLE-US-00035 EVQLLESGGGVVQPGRSLRLSCAASGFTFSTYGMHW (SEQ ID NO: 28)
VRQGPGKGLEWVSSISNSNTYIYYADAVKGRFTISR
DNAKNSLYLQMNSLRAEHTAVYYCARDSRYSNFLRW VRSDGMDVWGQGTTVIVSS.
[0247] The amino acid sequence of the anti-Rh(D) chain F01 is
TABLE-US-00036 AELTQSPSSLSASVGDRVTITCRASQGFRNDLGWYQ (SEQ ID NO: 29)
QKPGKAPKRLIYATSSLQSGVPSRFSGSGSGTEFTL
TINSLQPEDSATYYCLQHNSFPWTFGQGTKVEIKR.
[0248] The amino acid sequence of the anti-Rh(D) chain G01 is
TABLE-US-00037 AELTQSPLSLPVTPGEPASISCRSSQSLLHSSGFNF (SEQ ID NO: 30)
LDWYLQKPGQSPQLLIYMGSNRASGVPDRFSGSGSG
TDFTLKINRVEAEDVGVYYCMQALQFPLTFGGGTKV EIKR.
[0249] The amino acid sequence of the anti-Rh(D) chain H01 is
TABLE-US-00038 AELTQSPSFLSASVGDRVTITCRASQGITSYLAWYQ (SEQ ID NO: 31)
QKPGKAPKLLIYAASTLQSGVPSRFSGSGSGTEFTL
TIASLQPDDFATYYCQQLNNYPPFTFGPGTKVDIKR .
[0250] The amino acid sequence of the anti-Rh(D) chain I01 is
TABLE-US-00039 AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQ (SEQ ID NO: 32)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQSYSTPPYTFGQGTKLEIKR .
[0251] The amino acid sequence of the anti-Rh(D) chain I02 is
TABLE-US-00040 AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQ (SEQ ID NO: 33)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQSYSTLWTFGQGTKVEIKR.
[0252] The amino acid sequence of the anti-Rh(D) chain I03 is
TABLE-US-00041 AELTQSPSSLSASVADRVTITCRTSRNINRYLNWYQ (SEQ ID NO: 34)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTL
TITSLQPEDFATYYCQQSYSTPFTFGPGTKVDLKR.
[0253] The amino acid sequence of the anti-Rh(D) chain I04 is
TABLE-US-00042 AELTQSPSSLSASVGDRVTITCRASQNIRRSLNWYQ (SEQ ID NO: 35)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQSSNTPWTFGQGTKVEIKR.
[0254] The amino acid sequence of the anti-Rh(D) chain I05 is
TABLE-US-00043 AELTQSPSSLSASVGDRVTITCRASQSIRRYLNWYQ (SEQ ID NO: 36)
HKPGKAPKLLIFAASSLQSGVPSRFTGSGSGTDFTL
TISSLQPEDFATYYCQQSYSTPQTFGQGTKVEIKR.
[0255] The amino acid sequence of the anti-Rh(D) chain I06 is
TABLE-US-00044 AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQ (SEQ ID NO: 37)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQSYSTPITFGQGTRLEIKR.
[0256] The amino acid sequence of the anti-Rh(D) chain I07 is
TABLE-US-00045 AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQ (SEQ ID NO: 38)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQSYSTPRTFGGGTKVEIKR.
[0257] The amino acid sequence of the anti-Rh(D) chain I08 is
TABLE-US-00046 AELTQSPFSLSASVGDRVTITCRASQTISRSLNWYQ (SEQ ID NO:39)
HKPGEAPKLLIYAASSLQRGVPPRFSGSGSGTDFTL
TISSLQPEDFATYFCQQSYSTPPYSFGQGTKLEIKR .
[0258] The amino acid sequence of the anti-Rh(D) chain I09 is
TABLE-US-00047 AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQ (SEQ ID NO:40)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDSTL
TISSLQPEDFATYYCQQLNSYPYTFGQGTKLEIKR.
[0259] The amino acid sequence of the anti-Rh(D) chain I10 is
TABLE-US-00048 AELTQSPSSLSASVGDRVTITCRASQNISSYLNWYQ (SEQ ID NO:41)
QKPGKAPKLLIYAASSLQSGVLSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQSYTPPYSFGQGTKLEIKR.
[0260] The amino acid sequence of the anti-Rh(D) chain I11 is
TABLE-US-00049 AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQ (SEQ ID NO:42)
QKPGKAPTLLINAASSLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFAIYYCQQRETFGQGTKLEIKR.
[0261] The amino acid sequence of the anti-Rh(D) chain I12 is
TABLE-US-00050 AELTQSPSSLSASVGDRVTITCRASQSISSYLNWYQ (SEQ ID NO:43)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQSYSTPPYTFGQGTKLEIKR .
[0262] The amino acid sequence of the anti-Rh(D) chain I13 is
TABLE-US-00051 AELTQSPSSLSASVGDRVTITCRASQSISRYLNWYQ (SEQ ID NO: 44)
QKPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTL
TISSLQPEDFATYYCQQSYGTPHSFGRGTKLEIKR.
[0263] The amino acid sequence of the anti-Rh(D) chain I15 is
TABLE-US-00052 AELTQSPSSLSASVGDRVTITCRANQNIRRSLNWYQ (SEQ ID NO: 45)
QKPGKAPNLLIYAASTLQGGVPSRFSGSGSGTDFTL
TISSLQLADFATYYCQQTSATPWTFGQGTKVEIKR.
[0264] The amino acid sequence of the anti-Rh(D) chain I16 is
TABLE-US-00053 AELTQSPSSLPASVGDRVTITCRASQTIGFNLNWYQ (SEQ ID NO: 46)
QTSGKPPKLLIYGVSKLQNGVPSRFSGSGSGTEFTL
TISSLQPEDFATYYCQQTNDALWTFGQGTKVEVRR.
[0265] The amino acid sequence of the anti-Rh(D) chain J01 is
TABLE-US-00054 AELQDPVVSVALGQTVRITCQGDGLRSYYASWYQQK (SEQ ID NO: 47)
PGQAPKLVMYGRNNRPSGIPGRFSGSSSGQTAALTI
TGTQAEDEADYYCQSRATSGNPVVFGGGTKLTVL.
[0266] The amino acid sequence of the anti-Rh(D) chain J02 is
TABLE-US-00055 AELQDPVVSVALGQTVRITCQGDGLRSYYASWYQQK (SEQ ID NO: 48)
PGQAPKLVMYGRNNRPSGIPDRFSGSSSGQTAALTI
TGTQAEDEADYYCQSRATSGNPVVFGGGTKLTVL.
[0267] The amino acid sequence of the anti-Rh(D) chain J04 is
TABLE-US-00056 AELQDPVVSVALGQTVRITCQGDSLRSYYASWYQQK (SEQ ID NO:49)
PGQAPVLVIYGKNSRPSGIPDRFSGSSSGNTASLTI
TGAQAEDEADYYCSSRGSPHVAFGGGTKLTVL.
[0268] The amino acid sequence of the anti-Rh(D) chain J05 is
TABLE-US-00057 AELQDPVVSVALGQTVKITCQGDSLRKYYASWYQQK (SEQ ID NO:50)
PGQAPVLVFYARNSRPSGIPDRFSGSNSGTTASLTI
AGARAEDEADYYCHSRDSNGHHRVFGGGTKLTVL.
[0269] The amino acid sequence of the anti-Rh(D) chain K01 is
TABLE-US-00058 AELTQEPSLTVSPGGTVTLTCASSTGAVTSRYFPNW (SEQ ID NO: 51)
FQQKPGQAPRPLIYSASNKHSWTPARFSGSLLGGKA
ALTLSGVQPEDEAEYYCLLYYSGAWVFGGGTKLTVL .
[0270] The amino acid sequence of the anti-Rh(D) chain K02 is
TABLE-US-00059 AELTQEPSLTVSPGGTVTLTCASSTGAVTSRYFPNW (SEQ ID NO: 52)
FQQKPGQAPRPLIYSASNKHSWTPARFSGSLLGGKA
ALTLSGVQPEDEAEYYCLLYYSGAWVFGGGTKLTVL .
[0271] The amino acid sequence of the anti-Rh(D) chain K03 is
TABLE-US-00060 AELTQPPSLTVSPGGTVTLTCASSTGAVTSRYFPNW (SEQ ID NO:53)
FQQKPGQAPRALIYGSNNKHSWTPARFSGSLLGGKA ALTLSGVQPEDEAEYYCLLFYAGAWAFGGW
TKLTVL.
[0272] The amino acid sequence of the anti-Rh(D) chain L01 is
TABLE-US-00061 AELTQPPSASGTPGQRVTISCSGGSSNIASNTVNWY (SEQ ID NO:54)
QQLPGTAPKLLIYSNNQRPSGVPDRFSGSKSGTSAT
LVITGLQTGDEADYYCGTWDHSRSGAVFGGGTKLTV L.
[0273] The amino acid sequence of the anti-Rh(D) chain L03 is
TABLE-US-00062 AELTQPPSASGTPGQRVTISCSGSSSNIGNNHVSWY (SEQ ID NO:55)
QQLPGMAPKLLIYSNGQRPSGVPDRFSGSKSGTSAS
LAISGLQSEDEADYYCAAWHDSLYGPVFGGGTKLTV L.
[0274] The amino acid sequence of the anti-Rh(D) chain L04 is
TABLE-US-00063 AELTQPPSASGTPGQRVSISCSGSSSNIGSNTVNWY (SEQ ID NO:56)
QQLPGTAPKLLISTNNQGPSGVPDRFSGSKSGTSSS
LAISGLRSEAEDDYYCAAWDDTLNGVVFGGGTKLTV L.
[0275] The amino acid sequence of the anti-Rh(D) chain L05 is
TABLE-US-00064 AELTQPPSASGTPGLRVTISCSGSSSNIGSNIVNWY (SEQ ID NO:57)
QQLPGTAPKLLIFSNNKRPSGVPRFSGSKSGTSASL
AISGLQSEDEADYYCATWDDSLNGRVFGGGTKLTVL .
[0276] The amino acid sequence of the anti-Rh(D) chain M01 is
TABLE-US-00065 AELTQPPSASGTPGQRVTISCSGSNFNIGSNYVFWY (SEQ ID NO: 58)
QHVPGTAPKLLIYNNNQRPSGVPDRLSGSKSGASAS
LAINGLRSDDEADYYCTGWDDRLSGLIFGGGPKVTV L.
[0277] The amino acid sequence of the anti-Rh(D) chain M02 is
TABLE-US-00066 AELTQPPSASGTPGQRVTISCSGSSSNIGSNYVYWY (SEQ ID NO: 59)
QQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSAS
LAISGLRSEDEADYYCAAWDDSLSGWVFGGGTKLTV L.
[0278] The amino acid sequence of the anti-Rh(D) chain M03 is
TABLE-US-00067 AELTQPPSASGTPGQRVTISCSGSSSNIGSNYVYWY (SEQ ID NO: 60)
QQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSAS
QQLPGTAPKLLIYRNNQRPSGVPDRFSGSKSGTSAS
LAISGLRSEAEADYYCAAWDDSLSAVVFGGGTKLTV LL.
[0279] The amino acid sequence of the anti-Rh(D) chain N01 is
TABLE-US-00068 AELTQPPSVSAAPGQKVTISCSGSSSNIDSNYVSWY (SEQ ID NO: 61)
QQLPGTAPKLLIFDNYRRPSGIPDRFSGSKSGTSAT
LGITGLQTGDEADYYCATWDDSLNGRVFGGGTKLTV L.
[0280] The amino acid sequence of the anti-Rh(D) chain N02 is
TABLE-US-00069 AELTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWY (SEQ ID NO: 62)
QQLPGTAPKLLIYDNNKRPSGIPDRFSGSKSGTSAT
LGITGLQTGDEADYYCGTWDSSLSAGRVRRMFGGGT KLTVLG.
[0281] The amino acid sequence of the anti-Rh(D) chain O01 is
TABLE-US-00070 AELTQPPSVSGAPGQRVTISCTGSSSNIGAPYGVHW (SEQ ID NO: 63)
YQQFPGTAPKLVIYNDNNRPSGVPDRFSGSKSGTSA
SLAITGLQAEDEADYYCQSYDSSLSGRVFGGGTKLT VL.
[0282] The amino acid sequence of the anti-Rh(D) chain O02 is
TABLE-US-00071 AELTQPPSVSGAPGQTVTISCTGSSSSIGARYDVHW (SEQ ID NO: 64)
YQHLPGTAPKLLIYGNHNRPSGVPDRFSGSKSGTSA
SLAITGLQAEDEAEYYCQSYDNSLSGSSVFFGGGTK LTVL.
[0283] The amino acid sequence of the anti-Rh(D) chain O03 is
TABLE-US-00072 AELTQPPSGAPGQTVTISCTGSSSNIGAGYDVHWYQ (SEQ ID NO: 65)
QLPGTAPKLLIYGNSNRPSGVPDRFSGSKSGTSASL
AITGLQAEDEADYYCQSYDSSLSGPYVVFGGGTKLT VL.
[0284] The amino acid sequence of the anti-Rh(D) chain P01 is
TABLE-US-00073 AELTQPPSVSVAPRQTARITCGGDKIGSNTVHWYRQ (SEQ ID NO: 66)
MSGQAPVLVIYEDKKRPPGIPERFSGSTSGTTATLS
ISGAQVEDEADYYCYSRDNSGDQRRVFGAGTKLTVL .
[0285] The amino acid sequence of the anti-Rh(D) chain Q01 is
TABLE-US-00074 AELTQPPSATASLGGSVKLTCILQSGHRNYAVAWHH (SEQ ID NO: 67)
QEAGKGPRFLMTVTNDGRHIKGDGIPDRFSGSASGA
ERYLSISGLQSEDEGDYYCQTWGTGMHVFGGGTKLT VL.
[0286] The amino acid sequence of the anti-Rh(D) chain R01 is
TABLE-US-00075 AELTQPPSASGSPGQSVTISCTGASSDVGAYKHVSW (SEQ ID NO: 68)
YQQHPGKAPKLLTHEGTKRPSGVPDRFSGSKSGNTA
SLTVSGLQAEDEADYYCSSFAGNSVIFGGGTKLTVL .
[0287] The amino acid sequence of the anti-Rh(D) chain S01 is
TABLE-US-00076 AELTQPPSVSGSPGQSITISCSDVGNYNLVSWYQQY (SEQ ID NO: 69)
PGKAPKLIIYEGSKRPSGVSSRFSGSRSGNTASLTI
SGLQAEDEADYHCCSYAISSRIFGGGTKLTVL.
Nucleotide Sequences of Anti-Rh(D) Heavy and Light Chains
[0288] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain B01 is TABLE-US-00077
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 70)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCC
TCTGGATTCACCTTCAGGAGCTATGCTATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGCTACAGCATATGATGGAAAAAATAAATACTAC
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACGCTGTTTCTGCAAATGAAC
AGCCTGAGAGCTGAGGACACGGCTGTGTTTTACTGT
GCGAGAGGCGGATTTTACTATGATAGTAGTGGTTAT
TACGGCTTGAGGCACTACTTTGACTCCTGGGGCCAG GGAACCCTGGTCACCGTCTCCTCA.
[0289] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain CO is TABLE-US-00078
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 71)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCC
TCTGGATTCTCCTTCAGTAGCTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
TCAGTTATATCATATGATGGACATCATAAAAACTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAAAACGCTGTACCTGCAAATGAAC
AGCCTGAGACCTGAGGACACGGCTGTATATTACTGT
GCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCT
GTTCCCTTTGATATCTGGGGCCCAGGGACAATGGTC ACCGTCTCTTCA.
[0290] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain C03 is TABLE-US-00079
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGTGTGGTC (SEQ ID NO: 72)
CAGCATGGGAGGTCCCTGAGACTGTCCTGTGCAGCC
TCTGGATTCTCCTTCAGTAGCTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
TCAGTTATATCATATGATGGACATCATAAAAACTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAAAACGCTGTACCTGCAAATGAAC
AGCCTGAGACCTGAGGACACGGCTGTATATTACTGT
GCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCT
GTTCCCTTTGATATATGGGGCCCAGGGACAATGGTC ACCGTGTCTTCA.
[0291] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain C04 is TABLE-US-00080
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 73)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCC
TCTGGATTCTCCTTCAGTACCTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
TCAGTTATATCATATGATGGACATAATAAAAACTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAAAACGCTGTACCTGCAAATGAAC
AGCCTGAGACCTGAGGACACGGCTGTGTATTACTGT
GCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCT
ATTCCTTTTGATATCTGGGGCCAAGGGACAATGGTC ACCGTCTCTTCA.
[0292] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain C05 is TABLE-US-00081
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTC (SEQ ID NO: 74)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCC
TCTGGATTCAGCTTCAGTAGTTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGTTATATCGTATGATGGAACTAATAAATACTTT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAAAACGCTGTATCTGCAAATGACC
AGCCTGAGACCTGAGGACACGGCTGTGTATTTCTGT
GCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCC
GTACCTCTTGATATCTGGGGCCAAGGGACAATGGTC ACCGTCTCTTCA.
[0293] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain C08 is TABLE-US-00082
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTC (SEQ ID NO: 75)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCC
TCTGGATTCAGCTTCAGTAGTTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGTTATATCGTATGATGGAACTAATAAATACTTT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAAAACGCTGTATCTGCAAATGACC
AGCCTGAGACCTGAGGACACGGCTGTGTATTTCTGT
GCGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCT
GTACCTCTTGATATCTGGGGCCAAGGGACAATGGTC ACCGTCTCTTCA.
[0294] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain C10 is TABLE-US-00083
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 76)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCC
TCTGGATTCTCCTTCAGTAGCTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGTGGAGTGGGTGT
CAGTTATATCATATGATGGACATCATAAAAACTATG
CAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAG
ACAATTCCAAGAAAACGCTGTACCTGCAAATGAACA
GCCTGAGACCTGAGGACACGGCTGTATATTACTGTG
CGAACCTAAGGGGGGAAGTAACTCGTCGTGCGTCTG
TTCCCTTTGATATCTGGGGCCCAGGGACATTGGTCA CCGTCTCTTCA.
[0295] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D01 is TABLE-US-00084
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 77)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTG
TCTGGTTTCACCTTCAATAACTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGTTATTTGGTTTGATGGAAGTAATAAATACTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACACTGTACCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTATATTACTGT
GCGAGAGAGAACCAGATAAAGCTATGGTCCCGATAC
CTTTACTACTTTGATTACTGGGGCCAGGGAACCCTG GTCACCGTCTCCTCA.
[0296] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D03 is TABLE-US-00085
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 78)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCG
TCTGGATTCACCTTCAGTACCTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGACTGGAGTGGGTG
GCAGTTATATGGTTTGATGGAAGTAATAAGGAATAT
GCAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGA
GACAATTCCAAGAACACGCTGTATCTACAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGT
GCGAGAGAAGAAGTGGTTCGGGGAGTTATCTTATGG
TCTCGGAAGTTTGACTACTGGGGCCAGGGAACCCTG GTCACCGTCTCCTCA.
[0297] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D04 is TABLE-US-00086
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGCC (SEQ ID NO: 79)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGCG
TCTGGATTCAGCCTCAGGAGCTATGGCATGCACTGG
GTCCGCCAGGCTCCTGGCAAGGGGCTGGAGTGGGTG
GCAGATATATGGTTTGATGGAAGTAATAAAGATTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACGTTGTATCTTCAAATGAAC
AGCCTGAGAGCCGAGGATACGGCTGTGTATTATTGT
GCGAGAGATTGGAGGGTGCGGGCCTTTAGTAGTGGC
TGGTTAAGTGCTTTTGATATCTGGGGCCAAGGGACA ATGGTCACCGTCTCCTCA.
[0298] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D05 is TABLE-US-00087
GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCGTG (SEQ ID NO: 80)
GCCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTA
GCGTCTGGATTCAGCCTCAGGAGCTATGGCATGCAC
TGGGTCCGCCAGGCTCCTGGCAAGGGGCTGGAGTGG
GTGGCAGATATATGGTTTGATGGAAGTAATAAAGAT
TATGCAGACTCCGTGAAGGGCCGATTCACCATCTCC
AGAGACAATTCCAAGAACACGTTGTATCTTCAAATG
AACAGCCTGAGAGCCGAGGACACGGCTGTGTATTAT
TGTGCGAGAGATTGGAGGGTGCGGGCCTTTAGTAGT
GGCTGGTTAAGTGCTTTTGATATCTGGGGCCAAGGG ACCACGGTCAGCGTCTCCTCA.
[0299] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D07 is TABLE-US-00088
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 81)
CAGCCTGGGAGGTCCCTGAGAACTCTCCTGTGCAGT
GTCTGGATTCACCCTAACTAATTATGGCATGCACTG
GGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGT
GGCACATGTCTGGTATGATGGAAGTAAAACAGAATA
TGCAGACTCCGTCAAGGGCCGATTCGCCGTCTCCAG
AGACAAATCCAAGAACACACTGTTTCTGCAAATGAA
CAGCCTGACAGCCGAGGACACGGCTATTTATTACTG
TGCGAGAGAGAGGAGAGAGAAAGTCTATATATTGTT
CTACTCGTGGCTCGACCGCTGGGGCCAGGGAACCCT GGTCACCGTCTCCTCA.
[0300] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D08 is TABLE-US-00089
GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCGTG (SEQ ID NO: 82)
GTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCA
GCGTCTGGGTTCACCTTCAGTAGCTATGGCATGCAC
TGGGTCCGCCAGGCTCCAGGCAGGGGGCTGGAGTGG
GTGGCTCTTATATGGTACGATGGAGGTAACAAAGAG
TATGCAGACTCCGTGAAGGGCCGCTTCAGCATCTCC
AGAGACAATTCCAAGAACACTCTGTATCTGCAAGTG
AACAGCCTGAGAGCCGACGACACGGCTGTCTATTAC
TGTGCGAGAGACCAGAGAGCAGCAGCGGGTATCTTT
TATTATTCCCGTATGGACGTCTGGGGCCAAGGGACC ACGGTCACCGTCTCCTCA.
[0301] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D09 is TABLE-US-00090
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 83)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGAAGCG
TCTAAATTCACCCTCTACAATTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCATTTATATGGTTTGATGGAAGTAATAAATACTAT
GAAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGA
GACAATTCCAAGAACACGCTGTATCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGT
GCGAGAGAAGGATCTAAGAAGGTGGCACTTTCTAGG
TATTACTATTATATGGACGTCTGGGGCCAGGGGACC ACGGTCACTGTCTCGTCA.
[0302] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D10 is TABLE-US-00091
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 84)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGAAGCG
TCTAAATTCACCCTCTACAATTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCATTTATATGGTTTGATGGAAGTAATAAATACTAT
GAAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGA
GACAATTCCAAGAACACGCTGTATCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGT
GCGAGAGAAGTATCTAAGAAGGTGGCACTTTCTAGG
TATTACTACTATATGGACGTCTGGGGCCAGGGGACC ACGGTCACTGTCTCCTCA.
[0303] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D11 is TABLE-US-00092
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 85)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGAAGCG
TCTAAATTCACCCTCTACAATTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCGAAGGGCTGGAGTGGGTG
GCATTTATATGGTTTGATGGAAGTAATAAATACTAT
GCAGACTCCGTGAAGGGCCGATTCACCGTCTCCAGA
GACAATTCCAAGAACACGCTGTATCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGT
GCGAGAGAAGTATCTAAGAAGCTGGCACTTTCTAGG
TACTACTACTATATGGACGTCTGGGGCCAGGGGACC ACGGTCACTGTCTCCTCA.
[0304] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D12 is TABLE-US-00093
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTC (SEQ ID NO: 86)
CAGCCTGGGAGGTCCCTGAGACTCGCCTGTGCAGCG
TCTGGATTCAGCTTCAGGAGCTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAGGGGGCTGGAGTGGGTG
GCATTTACATGGTTTGATGGAAGCAATAAATATTAT
GTAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACGCTGTATCTGGAAATGAAC
AGCCTGAGAGTCGATGACACGGCTGTATATTACTGT
GCGAGAGAGGCGTCTATGCTTCGCGGAATTAGCAGA
TACTACTACGCGATGGACGTCTGGGGCCCAGGGACC ACGGTCACCGTCTCCTCA.
[0305] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D13 is TABLE-US-00094
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTC (SEQ ID NO: 87)
CAGCCTGGGAGGTCCCTGAGACTCGCCTGTGCAGCG
TCTGGATTCAGCTTCAGGAGCTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAGGGGGCTGGAGTGGGTG
GCATTTACATGGTTTGATGGAAGCAATAAATATTAT
GTAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACGCTGTATCTGGAAATGAAC
AGCCTGAGAGTCGATGACACGGCTGTATATTACTGT
GCGAGAGAGGGCGTCTATGCTTCGCGGAATTAGCAG
ATACTACTACGCGATGGACGTCTGGGGCCCAGGGAC CACGGTCACCGTCTCCTCA.
[0306] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D14 is TABLE-US-00095
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCTTGGTA (SEQ ID NO: 88)
CAGCCTGGGGGGTCCCTGAGACTCTCCTGTTGCAGC
GTCTGGATTCACCTTCAGTACTTATGGCATGCACTG
GGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGT
GGCAGTTATATGGTTTGATGGAAGTAAGAGAGACTA
TGCAGAGTCCGTGAAGGGCCGATTCACCATCTCCAG
AGACAACTCCAAGAACACACTGTATCTGCAAATGAA
CAGCCTGAGAGCCGAGGACTCGGCTGTGTATTACTG
TGCGAGAGAAAATGTGGCTCGTGGGGGGGGGGGCAT
TCGATACAAGTACTACTTTGACTACTGGGGCCAGGG AACCCTGGTCACCGTCTCCTCA.
[0307] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D15 is TABLE-US-00096
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 89)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTG
TCTGGATTCACCTTCAATAACTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGTTATTTGGTTTGATGGAAGTAATAAATACTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACACTGTACCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTATATTACTGT
GCGAGAGAGAACCAGATAAAGCTATGGTCCCGATAC
CTTTACTACTTTGACTACTGGGGCCAGGGAACCCTG GTCACCGTCTCCTCA.
[0308] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D16 is TABLE-US-00097
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 90)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTG
TCTGGTTTCACCTTCAATAACTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGTTATTTGGTTTGATGGAAGTAATAAATACTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACACTGTACCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTATATTACTGT
GCGAGAGAGAACCAGATAAAGCTATGGTCCCGATAC
CTTTACTACTTTGACTACTGGGGCCAGGGAACCCTG GTCACCGTCTCCTCA.
[0309] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D17 is TABLE-US-00098
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 91)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTG
TCTGGTTTCACCTTCAATAACTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGTTATTTGGTTTGATGGAAGTAATAAATACTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACACTGTACCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTATATTACTGT
GCGAGAGAGAACCAGATAAAGCTATGGTCCCGATAC
CTTTACTACTTTGACTACTGGGGCCAGGGAACCCTG GTCACCGTCTCCTCC.
[0310] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D18 is TABLE-US-00099
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 92)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGTAGTG
TCTGGTTTCACCTTCAATAACTATGGCATGCACTGG
GTCCGCCAGGCTTCAGGCAAGGGGTTGGAGTGGGTG
GCAGTTATTTGGTTTGATGGAAGTAATAAATACTAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACACTGTACCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTATATTACTGT
GCGAGAGAGAACCAGATAAAGCTATGGTCCCGATAC
CTTACTACTTTGACTACTGGGGCCAGGGAACCCTGG TCACCGTGTCCTCA.
[0311] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D20 is TABLE-US-00100
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTC (SEQ ID NO: 93)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCG
TCTGGATTCACCTTCAGTACCTATGGCATGCACTGG
GTCCGCCAGGCTCCAGGCAAGGGACTGGAGTGGGTG
GCAGTTATATGGTTTGATGGAAGTAATAAGGAATAT
GCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAATTCCAAGAACACGCTGTATCTACAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGT
GCGAGAGAAGAAGTGGTTCGGGGAGTTATCTTATGG
TCTCGGAAGTTTGACTACTGGGGCCAGGGAACCCTG GTCACCGTCTCCTCA.
[0312] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D30 is TABLE-US-00101
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGGTC (SEQ ID NO: 94)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCG
TCTGGATTCACCTTCAGTAGCTATGGCATGCGCTGG
GTCCGGCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGTTGTCTACTATGATGGAAGTAACAAACACTAT
TCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAACTCCAAGAACACGCTGTATCTACAAATGGAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGT
GCGAGAGAAAGAAATTTTCGGAGTGGTTATTCCCGC
TACTACTACGGTATGGACGTCTGGGGCCCAGGGACC ACGGTCACCGTCTCCTCA.
[0313] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain D31 is TABLE-US-00102
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGGTC (SEQ ID NO: 95)
CAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCG
TCTGGATTCACCTTCAGTAGCTATGGCATGCACTGG
GTCCGGCAGGCTCCAGGCAAGGGGCTGGAGTGGGTG
GCAGTTGTCTACTATGATGGAAGTAACAAACACTAT
TCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGA
GACAACTCCAAGAACACGCTGTATCTACAAATGGAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGT
GCGAGAGAAAGAAATTTTCGGAGTGGTTATTCCCGC
TACTACTACGGTATGGACGTCTGGGGCCCAGGGACC ACGGTCACCGTCTCCTCA.
[0314] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain E01 is TABLE-US-00103
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCCTGGTC (SEQ ID NO:96)
AAGCCTGGGGGGTCCTGAGACTCTCCTGTGCAGCCT
CTGGATTCACCTTCAGTAGCTATAGCATGCACTGGG
TCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCT
CATCCATTAGTAATAGTAATACTTACATATACTACG
CAGACGCAGTGAAGGGCCGATTCACCATCTCCAGAG
ACAACGCCAAGAACTCACTGTATCTGCAAATGAACA
GCCTGAGAGCCGAGGACACGGCTGTGTACTACTGTG
CGAGAGATTCTAGATACAGTAATTTCCTCCGTTGGG
TTCGGAGCGACGGTATGGACGTCTGGGGCCAAGGGA CCACGGTCATCGTCTCCTCA.
[0315] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain E03 is TABLE-US-00104
GAGGTGCAGCTGCTCGAGTCTGGGGTGGAGTCTGGG (SEQ ID NO:97)
GGAGGCCTGGTCAAGCCTGGGGGGTCCCTGAGACTC
TCCTGTGCAGCCTCTGGATTCACCTTCAGTAGCTAT
AGCATGCACTGGGTCCGCCAGGGTCCAGGGAAGGGG
CTGGAGTGGGTCTCATCCATTAGTAATAGTAATACT
TACATATACTACGCAGACGCAGTGAAGGGCCGATTC
ACCATCTCCAGAGACAACGCCAAGAACTCACTGTAT
CTGCAAATGAACAGCCTGAGAGCCGAGCACACGGCT
GTGTACTACTGTGCGAGAGATTCTAGATACAGTAAT
TTCCTCCGTTGGGTTCGGAGCGACGGTATGGACGTC
TGGGGCCAAGGGACCACGGTCATCGTCTCCTCA.
[0316] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain F01 is TABLE-US-00105
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTGCA (SEQ ID NO:98)
TCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCA
AGTCAGGGCTTTAGAAATGATTTAGGCTGGTATCAG
CAGAAACCAGGGAAAGCCCCTAAGCGCCTGATCTAT
GCTACATCCAGTTTGCAAAGTGGGGTCCCATCAAGG
TTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTC
ACAATCAACAGCCTGCAGCCTGAAGATTCTGCAACT
TATTACTGTCTACAGCATAATAGTTTCCCGTGGACG
TTCGGCCAAGGGACCAAGGTGGAAATCAAACGA.
[0317] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain G01 is TABLE-US-00106
GCCGAGCTCACTCAGTCTCCACTCTCCCTGCCCGTC (SEQ ID NO:99)
ACCCCTGGAGAGCCGGCCTCCATCTCCTGCAGGTCT
AGTCAGAGCCTCCTGCATAGTAGTGGATTCAACTTT
TTGGATTGGTACCTGCAGAAGCCAGGGCAGTCTCCA
CAGCTCCTGATCTATATGGGTTCTAATCGGGCCTCC
GGGGTCCCTGACAGGTTCAGTGGCAGTGGATCAGGC
ACAGATTTACACTGAAAATCAACAGAGTGGAGGCTG
AGGATGTTGGGGTTTATTACTGCATGCAAGCTCTAC
AATTTCCTCTCACTTTCGGCGGAGGGACCAAGGTGG AGATCAAACGA.
[0318] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain H01 is TABLE-US-00107
GCCGAGCTCACCCAGTCTCCATCCTTCCTGTCTG (SEQ ID NO:100)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCCAGTCAGGGCATTACGAGTTATTTAGCCTGG
TATCAGCAAAAACCAGGGAAAGCCCCTAAGCTCC
TAATCTATGCTGCATCCACTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGCGGCAGTGGATCTGGGACA
GAATTCACTCTCACAATCGCCAGCCTGCAGCCTG
ATGATTTTGCAACTTATTACTGTCAACAGCTTAA
TAATTACCCCCCTTTCACTTTCGGCCCTGGGACC AAAGTGGATATCAAACGA.
[0319] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I01 is TABLE-US-00108
GCCGAGCTCACCCAGTCTCCATCCTCCCTATCTG (SEQ ID NO:101)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCTCCGTACACTTTTGGCCAGGGGACC AAGCTGGAGATCAAACGA.
[0320] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I02 is TABLE-US-00109
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:102)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCTGTGGACGTTCGGCCAAGGGACCAAG GTGGAAATCAAACGA.
[0321] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I03 is TABLE-US-00110
GCCGAGCTCACCCAGTCTCCATCCTTCCTGTCTG (SEQ ID NO:103)
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG
CATCTGTAGCGGACAGAGTCACCATCACTTGCCG
GACAAGTCGGAACATTAACAGATACTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATTTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCACCAGTCTGCAACCTG
AAGATTTGCCACTTACTACTGTCAACAGAGTTAC
AGTACCCCTTTCACTTTCGGCCCTGGGACCAAAG TGGATCTCAACGA.
[0322] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I04 is TABLE-US-00111
GCCGAGCTCACTCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:104)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAACATTAGGAGGTCTTTAAATTGG
TATCAACAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGTC
CCATCAAGGTTCAGTGGCAGTGGATCTGGGACAG
ATTTCACTCTCACCATCAGCAGTCTGCAACCTGA
AGATTTTGCAACTTACTACTGTCAGCAGAGTTCC
AATACCCCGTGGACGTTCGGCCAAGGGACCAAGG TGGAAATCAAACGA.
[0323] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I05 is TABLE-US-00112
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:105)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGGAGGTATTTAAATTGG
TATCAGCACAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTTTGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCACTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCTCAAACGTTCGGCCAAGGGACCAAG GTGGAAATCAAACGA.
[0324] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I06 is TABLE-US-00113
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:106)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCCGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCGATCACCTTCGGCCAAGGGACACGA CTGGAGATTAAACGA.
[0325] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I07 is TABLE-US-00114
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:107)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCTCGAACTTTCGGCGGAGGGACCAAG GTGGAGATCAAACGA.
[0326] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I08 is TABLE-US-00115
GCCGAGCTCACCCAGTCTCCATTCTCCCTGTCTG (SEQ ID NO:108)
CATCTGTCGGAGACAGAGTCACCATAACTTGCCG
GGCAAGTCAGACCATTAGCAGGTCTTTAAATTGG
TATCAGCATAAACCAGGGGAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTCTGCAGCGTGGGGT
CCCACCCAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGACTTTGCGACTTACTTCTGTCAACAGAGTGT
CAGAATCCCGTACAGTTTTGGCCAGGGGACCAAG CTGGAGATCAAACGA.
[0327] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I09 is TABLE-US-00116
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:109)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTCCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTATTACTGTCAACAGCTTAA
TAGTTACCCGTACACTTTTGGCCAGGGGACCAAG CTGGAGATCAAACGA.
[0328] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I10 is TABLE-US-00117
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:110)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAACATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCTATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCTCCGTATAGTTTTGGCCAGGGGACC AAGCTGGAGATCAAACGA.
[0329] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I11 is TABLE-US-00118
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:111)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTACGCTCC
TGATCAATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATTAGCAGTCTGCAACCTG
AAGATTTCGCAATTTACTACTGTCAACAGAGAGA
AACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA CGA.
[0330] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I12 is TABLE-US-00119
GCCGAGCTCACCCAGTCTCCATCCTCCCTATCTG (SEQ ID NO:112)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCTCCGTACACTTTTGGCCAGGGGACC AAGCTGGAGATCAAACGA.
[0331] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I13 is TABLE-US-00120
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:113)
CCTCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGGTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCTAAGCTCCT
GATCTATGCTGCATCCAGTTTGCAAAGTGGGGTC
CCATCAAGGTTCAGTGGCAGTGGATCTGGGACAG
ATTTCACTCTCACCATCAGCAGTCTGCAACCTGA
AGATTTTGCAACTTACTACTGTCAACAGAGTTAC
GGTACCCCTCACAGTTTTGGCCGGGGGACCAAGC TGGAGATCAAACGA.
[0332] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I15 is TABLE-US-00121
GCCGAGCTCACCCAGTCTCCTTCCTCCCTGTCTG (SEQ ID NO:114)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAATCAGAACATTCGTAGATCTTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAACCTCC
TGATCTATGCTGCATCCACATTGCAAGGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACTTG
CGGATTTTGCAACTTACTACTGTCAACAGACTTC
CGCTACCCCGTGGACGTTCGGCCAAGGGACCAAG GTGGAAATCAAACGA.
[0333] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain I16 is TABLE-US-00122
GCCGAGCTCACCCAGTCTCCATCGTCCCTGCCTG (SEQ ID NO:115)
CATCTGTGGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGACTATTGGTTTTAATTTAAATTGG
TATCAGCAAACATCTGGGAAGCCCCCTAAACTCC
TAATCTATGGTGTTTCCAAGTTGCAAAATGGGGT
CCTTCACGGTTCAGTGGCAGTGGGTCCGGGACGG
AATTCACCCTCACAATCAGCAGTCTGCAGCCTGA
GGATTTTGCGACTTATTATTGTCAACAGACTAAC
GATGCGTTGTGGACGTTCGGCCAAGGGACCAAAG TGGAAGTCAGACGA.
[0334] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain J01 is TABLE-US-00123
GCCGAGCTCCAGGACCCTGTTGTGTCTGTGGCCT (SEQ ID NO:116)
TGGGACAGACAGTCAGGATCACTTGCCAAGGAGA
CGGCCTCAGAAGTTATTATGCAAGCTGGTACCAG
CAGAAGCCGGGACAGGCCCCGAAACTTGTCATGT
ACGGTAGAAACAACCGGCCCTCAGGGATCCCAGG
CCGATTCTCTGGCTCCAGCTCAGGGCAGACAGCT
GCCTTGACCATCACGGGGACTCAGGCGGAGGATG
AGGCTGACTATTACTGTCAGTCCCGTGCCACCAG
CGGTAACCCTGTGGTGTTCGGCGGAGGGACTAAG CTGACCGTCCTG.
[0335] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain J02 is TABLE-US-00124
GCCGAGCTCCAGGACCCTGTTGTGTCTGTGGCCT (SEQ ID NO:117)
TGGGACAGACAGTCAGGATCACTTGCCAAGGAGA
CGGCCTCAGAAGTTATTATGCAAGCTGGTACCAG
CAGAAGCCGGGACAGGCCCCGAAACTTGTCATGT
ACGGTAGAAACAACCGGCCCTCAGGGATCCCAGA
CCGATTCTCTGGCTCCAGCTCAGGGCAGACAGCT
GCCTTGACCATCACGGGGACTCAGGCGGAGGATG
AGGCTGACTATTACTGTCAGTCCCGTGCCACCAG
CGGTAACCCTGTGGTGTTCGGCGGAGGGACTAAG CTGACCGTCCTG.
[0336] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain J04 is TABLE-US-00125
GCCGAGCTCCAGGACCCTGTTGTGTCTGTGGCCT (SEQ ID NO:118)
TGGGACAGACAGTCAGGATCACATGCCAAGGAGA
CAGCCTCAGAAGCTATTATGCAAGCTGGTACCAG
CAGAAGCCAGGACAGGCCCCTGTACTTGTCATCT
ATGGTAAAAACAGCCGGCCCTCAGGGATCCCAGA
CCGATTCTCTGGCTCCAGCTCAGGAAACACAGCT
TCGTTGACCATCACTGGGGCTCAGGCGGAAGATG
AGGCGGACTATTATTGTAGTTCGCGGGGCAGCCC
CCACGTGGCATTCGGCGGAGGGACCAAACTGACC GTCCTG.
[0337] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain J05 is TABLE-US-00126
GCCGAGCTCCAGGACCCTGTTGTGTCTGTGGCCT (SEQ ID NO:119)
TGGGACAGACAGTCAAGATCACATGCCAGGGAGA
CAGCCTCAGAAAGTATTATGCAAGCTGGTACCAG
CAGAAGCCAGGACAGGCCCCTGTGCTTGTCTTCT
ATGCTAGAAATAGCCGGCCCTCAGGGATCCCAGA
CCGATTCTCTGGCTCCAACTCAGGAACCACAGCT
TCCTTGACCATCGCTGGGGCTCGGGCGGAAGATG
AGGCTGACTATTACTGTCACTCCCGGGACAGCAA
TGGTCACCATCGGGTGTTCGGCGGAGGGACCAAG CTGACCGTCCTA.
[0338] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain K01 is TABLE-US-00127
GCCGAGCTCACTCAGGAGCCCTCACTGACTGTGT (SEQ ID NO:120)
CCCCAGGAGGGACAGTCACTCTCACCTGTGCTTC
CAGCACTGGAGCAGTCACCAGTCGTTACTTTCCA
AACTGGTTCCAGCAGAAACCTGGACAAGCACCCA
GGCCACTGATTTATAGTGCAAGCAACAAACACTC
CTGGACCCCTGCCCGGTTCTCAGGCTCCCTCCTT
GGGGGCAAAGCTGCCCTGACACTGTCAGGTGTGC
AGCCTGAGGACGAGGCTGAGTATTACTGCCTGCT
CTACTATAGTGGTGCTTGGGTGTTCGGCGGAGGG ACCAAGTTGACCGTCCTT.
[0339] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain K02 is TABLE-US-00128
GCCGAGCTCACTCAGGAGCCCTCACTGACTGTGT (SEQ ID NO:121)
CCCCAGGAGGGACAGTCACTCTCACCTGTGCTTC
CAGCACTGGAGCAGTCACCAGTCGTTACTTTCCA
AACTGGTTCCAGCAGAAACCTGGACAAGCACCCA
GGCCACTGATTTATAGTGCAAGCAACAAACACTC
CTGGACCCCTGCCCGGTTCTCAGGCTCCCTCCTT
GGGGGCAAAAGCTGCCCTGACACTGTCAGGTGTG
CAGCCTGAGGACGAGGCTGAGTATTACTGCCTGC
TCTACTATAGTGGTGCTTGGGTGTTCGGCGGAGG GACCAAGCTGACCGTCCTA.
[0340] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain K03 is TABLE-US-00129
GCCGAGCTCACTCAGCCACCCTCACTGACTGTGT (SEQ ID NO:122)
CCCCAGGAGGGACAGTCACTCTCACCTGTGCTTC
CAGCACTGGAGCAGTCACCAGTCGTTACTTTCCA
AACTGGTTCCAGCAGAAACCTGGCCAGGCACCCA
GGGCACTGATTTATGGTTCAAACAACAAACACTC
CTGGACCCCTGCCCGGTTCTCAGGCTCCCTCCTT
GGGGGCAAAGCTGCCCTGACACTGTCAGGTGTGC
AGCCTGAGGACGAGGCGGAGTATTACTGCCTGCT
CTTCTATGCTGGTGCTTGGGCGTTCGGCGGATGG ACCAAGCTGACCGTCCTA.
[0341] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain L01 is TABLE-US-00130
CCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGG (SEQ ID NO:123)
AGGCAGCTCCAACATCGCAAGTAATACTGTAAAC
TGGTACCAGCAACTCCCAGGAACGGCCCCCAAAC
TCCTCATCTATAGTAATAATCAGCGGCCCTCAGG
GGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGC
ACCTCAGCCACCCTGGTCATCACCGGGCTCCAGA
CTGGGGACGAGGCCGATTATTACTGCGGAACATG
GGATCACAGCCGGAGTGGTGCGGTGTTCGGCGGA GGGACCAAACTGACCGTCTTA.
[0342] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain L03 is TABLE-US-00131
GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGA (SEQ ID NO:124)
CCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGG
CAGTAGCTCCAACATCGGAAATAATCATGTAAGC
TGGTACCAGCAACTCCCAGGAATGGCCCCCAAAC
TCCTCATCTATTCTAATGGTCAGCGGCCCTCAGG
GGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGC
ACCTCAGCCTCCCTGGCCATCAGCGGCCTCCAGT
CTGAGGATGAGGCTGATTATTATTGTGCAGCATG
GCATGACAGCCTCTATGGTCCGGTGTTCGGCGGA GGGACCAAGCTGACCGTCCTC.
[0343] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain L04 is TABLE-US-00132
GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGA (SEQ ID NO:125)
CCCCCGGGCAGAGGGTCAGCATCTCTTGTTCTGG
AAGCAGCTCCAACATCGGAAGTAATACTGTAAAC
TGGTACCAGCAGCTCCCAGGAACAGCCCCCAAAC
TCCTCATCTCTACTAATAATCAGGGGCCCTCAGG
AGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGC
ACCTCATCCTCCCTGGCCATCAGTGGGCTCCGGT
CAGAGGCTGAGGATGATTATTACTGTGCAGCATG
GGATGACACCCTGAATGGTGTGGTATTCGGCGGA GGGACCAAACTGACCGTCCTA.
[0344] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain L05 is TABLE-US-00133
GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGA (SEQ ID NO:126)
CTCCCGGGCTGAGGGTCACCATCTCTTGTTCTGG
AAGCAGCTCCAACATCGGAAGTAATATTGTAAAC
TGGTACCAGCAGCTCCCAGGAACGGCCCCCAAAC
TCCTCATCTTTAGTAATAATAAGCGGCCCTCAGG
GGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGC
ACCTCAGCCTCCCTGGCCATCAGTGGGCTCCAGT
CTGAGGATGAGGCTGATTATTACTGTGCTACATG
GGATGACAGCCTGAATGGTCGGGTGTTCGGCGGA GGGACCAAGCTGACCGTCCTA.
[0345] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain M01 is TABLE-US-00134
GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGA (SEQ ID NO:127)
CCCCCGGGCAGCGGGTCACCATCTCTTGTTCTGG
GAGCAACTTCAACATCGGAAGTAATTATGTATTC
TGGTACCAGCATGTTCCAGGAACGGCCCCAAAAC
TCCTCATCTATAATAATAATCAACGCCCCTCTGG
GGTCCCTGACCGACTCTCTGGCTCCAAGTCTGGC
GCCTCAGCCTCCCTGGCCATCAATGGGCTCCGGT
CCGATGATGAGGCTGATTATTACTGTACAGGATG
GGATGACCGCCTGAGTGGCCTGATTTTCGGCGGA GGGCCAAAAGTGACCGTCCTA.
[0346] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain M02 is TABLE-US-00135
GCCGAGCTCACGCAGCCGCCCTCAGCGTCTGGGA (SEQ ID NO:128)
CCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGG
AAGCAGCTCCAACATCGGAAGTAATTATGTATAT
TGGTACCAGCAGCTCCCAGGAACGGCCCCCAAAC
TCCTCATCTATAGGAATAATCAGCGGCCCTCAGG
GGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGC
ACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGT
CCGAGGATGAGGCTGATTATTACTGTGCAGCATG
GGATGACAGCCTGAGTGGTTGGGTGTTCGGCGGA GGGACCAAGCTGACCGTCCTA.
[0347] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain M03 is TABLE-US-00136
GCCGAGCTCACTCAGCCACCCTCAGCGTCTGGGA (SEQ ID NO:129)
CCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGG
AAGCAGCTCCAACATCGGAAGTAATTATGTATAC
TGGTACCAGCAGCTCCCAGGAACGGCCCCCAAAC
TCCTCATCTATAGGAATAATCAGCGGCCCTCAGG
GGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGC
ACCTCAGCCTCCCTGGCCATCAGTGGGCTCCGGT
CCGAGGCTGAGGCTGATTATTACTGTGCGGCATG
GGATGACAGCCTGAGTGCCGTGGTATTCGGCGGA GGGACCAAACTGACCGTCCTA.
[0348] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain N01 is TABLE-US-00137
GCCGAGCTCACGCAGCCGCCCTCAGTGTCTGCGG (SEQ ID NO:130)
CCCCAGGACAGAAGGTCACCATCTCCTGCTCTGG
AAGCAGCTCCAACATTGACAGTAACTATGTATCC
TGGTACCAGCAGCTCCCAGGAACAGCCCCCAAAC
TCCTCATTTTTGACAATTATAGGCGACCCTCAGG
GATTCCTGACCGATTCTCAGGCTCCAAGTCTGGC
ACGTCAGCCACCCTGGGCATCACCGGACTCCAGA
CTGGGGACGAGGCCGATTATTACTGTGCAACATG
GGATGACAGCCTGAATGGTCGGGTGTTCGGCGGA GGGACCAAGCTGACCGTCCTA.
[0349] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain N02 is TABLE-US-00138
GCCGAGCTCACGCAGCCGCCCTCAGTGTCTGCGG (SEQ ID NO:131)
CCCCAGGACAGAAGGTCACCATCTCCTGCTCTGG
AAGCAGCTCCAACATTGGGAATAATTATGTGTCC
TGGTACCAGCAACTCCCAGGAACAGCCCCCAAAC
TCCTCATTTATGACAATAATAAGCGACCCTCAGG
GATTCCTGACCGATTCTCTGGCTCCAAGTCTGGC
ACGTCAGCCACCCTGGGCATCACCGGACTCCAGA
CTGGGGACGAGGCCGATTATTACTGCGGAACATG
GGATAGCAGCCTGAGTGCTGGCCGCGTTCGGCGG
ATGTTCGGCGGAGGGACCAAGTTGACCGTCCTGG GT.
[0350] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain O01 is TABLE-US-00139
GCCGAGCTCACGCAGCCGCCCTCAGTGTCTGGGG (SEQ ID NO:132)
CCCCAGGGCAGAGGGTCACCATCTCCTGCACTGG
GAGCAGCTCCAACATCGGGGCACCTTATGGTGTA
CACTGGTACCAGCAGTTTCCAGGAACAGCCCCCA
AACTCGTCATCTACAATGACAACAATCGGCCCTC
AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCT
GGCACCTCAGCCTCCCTGGCCATCACTGGGCTCC
AGGCTGAGGATGAGGCTGATTATTACTGCCAGTC
CTATGACAGCAGCCTGAGTGGAAGGGTGTTCGGC GGAGGGACCAAGCTGACCGTCCTA.
[0351] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain O02 is TABLE-US-00140
GCCGAGCTCACGCAGCCGCCCTCAGTGTCTGGGG (SEQ ID NO:133)
CCCCAGGGCAGACGGTCACCATCTCCTGCACTGG
GAGCAGCTCCAGCATCGGGGCACGTTATGATGTA
CACTGGTACCAACACCTTCCAGGAACAGCCCCCA
AACTCCTCATCTATGGTAACCACAATCGGCCCTC
AGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCT
GGCACCTCAGCCTCCCTGGCCATCACTGGGCTCC
AGGCTGAGGATGAGGCTGAATATTATTGCCAGTC
CTATGACAACAGCCTGAGTGGTTCGTCTGTCTTT
TTCGGCGGAGGGACCAAGCTGACCGTCCTA.
[0352] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain O03 is TABLE-US-00141
GCCAGACGGTCACCATCTCCTGCACTGGGAGCAG (SEQ ID NO:134)
CTCCAACATCGGGGCAGGTTATGATGTACACTGG
TACCAGCAGCTTCCAGGAACAGCCCCCAAACTCC
TCATCTATGGTAACAGCAATCGGCCCTCAGGGGT
CCCTGACCGATTCTCTGGCTCCAAGTCTGGCACC
TCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTG
AGGATGAGGCTGATTATTACTGCCAGTCCTATGA
CAGCAGCCTGAGTGGTCCCTATGTGGTATTCGGC GGAGGGACCAAGCTGACCGTCCTA.
[0353] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain P01 is TABLE-US-00142
GCCGAGCTCACTCAGCCACCCTCGGTGTCAGTGG (SEQ ID NO:135)
CCCCAAGACAGACGGCCAGGATTACCTGTGGGGG
GGACAAAATCGGAAGTAACACTGTGCATTGGTAC
CGGCAGATGTCAGGCCAGGCCCCTGTTCTGGTCA
TCTATGAAGACAAAAAACGACCCCCCGGGATCCC
TGAGAGATTCTCTGGTTCCACCTCAGGGACAACG
GCCACCTTGAGTATCAGTGGGGCCCAGGTTGAGG
ATGAAGCTGACTACTACTGTTATTCAAGAGACAA
CAGTGGTGATCAGAGAAGGGTGTTCGGCGCAGGG ACCAAGCTGACCGTCCTA.
[0354] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain Q01 is TABLE-US-00143
GCCGAGCTCACTCAGCCACCCTCCGCCACTGCCT (SEQ ID NO:136)
CCCTGGGAGGCTCGGTCAAACTCACCTGCATTCT
GCAGAGTGGCCACAGAAATTACGCCGTCGCTTGG
CATCACCAAGAAGCAGGGAAGGGCCCGCGATTTT
TGATGACGGTTACCAATGATGGCAGGCACATCAA
GGGGGACGGGATCCCTGATCGCTTCTCAGGCTCC
GCCTCTGGGGCTGAACGCTACCTCTCCATCTCCG
GCCTCCAGTCTGAGGATGAGGGTGACTACTACTG
TCAGACCTGGGGCACTGGCATGCATGTGTTCGGC GGAGGGACCAAACTGACCGTCCTA.
[0355] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain R01 is TABLE-US-00144
GCCGAGCTCACTCAGCCTCCCTCCGCGTCCGGGT (SEQ ID NO:137)
CTCCTGGACAGTCAGTCACCATCTCCTGCACTGG
AGCCAGCAGTGACGTTGGTGCTTATAAGCACGTC
TCCTGGTACCAACAACACCCAGGCAAAGCCCCCA
AACTCCTGACTCATGAGGGCACTAAGCGGCCCTC
AGGGGTCCCTGATCGCTTCTCTGGCTCCAAGTCT
GGCAACACGGCCTCCCTGACCGTCTCTGGGCTCC
AGGCTGAGGATGAGGCTGATTATTACTGCAGCTC
ATTTGCAGGTAATTCCGTGATATTCGGCGGAGGG ACCAAGCTGACCGTCCTA.
[0356] The nucleotide sequence of the portion of the clone encoding
the anti-Rh(D) chain S01 is TABLE-US-00145
GCCGAGCTCACTCAGCCTCCCTCCGTGTCTGGGT (SEQ ID NO:138)
CTCCTGGACAGTCGATCACCATCTCCTGCAGTGA
TGTTGGGAATTATAACCTTGTCTCCTGGTACCAA
CAGTACCCAGGCAAGGCCCCCAAACTCATAATTT
ATGAGGGCAGTAAGCGGCCCTCAGGGGTTTCTAG
TCGCTTCTCTGGCTCCAGGTCTGGCAACACGGCC
TCCCTGACAATCTCTGGGCTCCAGGCTGAGGACG
AGGCTGATTATCACTGCTGCTCATATGCAATTAG
TAGCAGGATTTTCGGCGGAGGGACCAAGCTGACC GTCCTA.
EXAMPLE 3
Isolation of Anti-Rh(D) Monoclonal Antibodies to Conventional and
Novel Epitopes Using a Heavy Chain/Light Chain Shuffling
Approach
[0357] In view of the results obtained in Examples 1 and 2 herein,
heavy and light chains of antibodies of various Rh(D) epitope
specificities were randomly recombined in order to generate
anti-Rh(D) antibodies having additional patterns of reactivity with
Rh(D) variant cells. Using this approach, plasmid DNA obtained from
the Fab/phage display libraries described in panning rounds 2 and 3
of Example 1 was randomly recombined to generate a "shuffled"
Fab/phage display library. When the Rh(D) specificity of antibodies
of this "shuffled" library was determine, it was found that many of
these antibodies exhibited novel epitope specificity.
Significantly, antibody clones having novel Rh(D) epitope
specificity were identified, including clones which bind to wild
type and certain partial D type red blood cells but which do not
bind to D category III red blood cells. The experiments described
in this Example therefore demonstrate that the methods described in
this specification may be used to generate antibody clones useful
for diagnostic and therapeutic applications in humans.
[0358] The materials and methods used in the experiments described
in this Example are now described.
Creation of Shuffled Fab/Phage Display Library
[0359] Two microgram aliquots of DNA obtained from libraries LP2,
LP3, KP2, and KP3 (described herein in Example 1) were digested
using the restriction endonucleases Spel and XhoI (15 and 60 units,
respectively) in order to dissociate DNA segments encoding
individual (full length) heavy chains from library plasmids
encoding individual (full length) light chains. Endonuclease/DNA
mixtures were incubated overnight at 37.degree. C. After the
restriction endonucleases were removed using standard
phenol/chloroform and chloroform extraction techniques, the DNA was
precipitated using ethanol.
[0360] Equivalent amounts of DNA from each of the four libraries
(500 nanograms total) were mixed, and then the heavy chain-encoding
DNA fragments were re-ligated into the library plasmids encoding
individual light chains. This ligation was performed overnight at
20.degree. C. in the presence of 3.5 units of T4 DNA ligase in a
total reaction volume of 70 microliters. This treatment generated
re-ligated library plasmids encoding a light chain and a heavy
chain, wherein the light chain and the heavy chain were not
necessarily encoded by a single plasmid in the original library
DNA. For this reason, the library of re-ligated plasmids was
designated a "shuffled" library.
[0361] Three microliters of shuffled library suspension were mixed
with an aliquot of XL1-Blue electrocompetent cells (obtained from
Stratagene, La Jolla, Calif.), and the cells were electroporated
according to standard methods. Electroporated cells were cultured
on plates containing Luria broth comprising 100 micrograms per
milliliter carbenicillin.
Anti-Rh(D) Specificity of "Shuffled" Library Antibodies
[0362] Fifty-six randomly chosen colonies were selected, and
monoclonal Fab/phage preparations were separately produced from
each of these individual colonies, using the methods described
herein in Example 1. Rh(D) specificity was determined by indirect
agglutination using anti-M13 antibody, as described herein in
Examples 1 and 2. Plasmid DNA was separately prepared from each of
the Fab/phage preparations which exhibited Rh(D) specificity, and
the DNA sequences encoding the heavy and light chains expressed by
each preparation were determined as described herein.
[0363] The results of the experiments presented in this Example are
now described.
Anti-Rh(D) Specificity of "Shuffled" Library Antibodies
[0364] Of the 56 randomly-chosen "shuffled" library clones, 34
(61%) demonstrated specificity for Rh(D). The Rh(D) epitope
specificity, the agglutination pattern, and the heavy and light
chain sequences of these 34 clones are listed in Table 4. Of these
34 clones, 19 exhibited specificity for previously-described Rh(D)
epitopes (e.g. epD 1, epD 2, epD 6/7, and epD X), and one bound too
weakly to wild-type Rh(D)-positive red blood cells to characterize
is epitope specificity (i.e. clone SH44). However, 14 of the clones
identified in Table 4 exhibited novel Rh(D) epitope specificity.
Some of these 14 antibody clones comprised a heavy chain, a light
chain, or both, that were identified herein in Examples 1 or 2.
However, half (17/34) of the heavy chain sequences and about 80%
(28/34) of the light chain sequences had not been identified in
Examples 1 or 2.
[0365] The Rh(D)-specific antibody clones isolated from the
"shuffled" library are useful for characterizing and classifying
patient red blood cells that express variant forms of the Rh(D)
antigen. Of particular interest are clones SH18, SH20, and SH46.
These three clones agglutinate wild type red blood cells and
certain partial D-type red blood cells, but do not agglutinate D
category III red blood cells (a.k.a partial Rh(D)III cells). It is
believed that all previously-characterized human monoclonal
anti-Rh(D) antibodies agglutinate D category III red blood cells.
Therefore these three clones are particularly useful for
differentiating D category III red blood cells from other types of
red blood cells.
[0366] From a clinical perspective, it has heretofore only been
possible to retrospectively identify D category III red blood cells
in a patient after they have been erroneously presumed to have
wild-type Rh(D)-positive cells. For example, transfusion of an
individual having D category III red blood cells with wild-type
Rh(D) cells induces production of anti-Rh(D) alloantibodies in the
individual. Previously, the presence of D category III red blood
cells in patients could only be determined by the production of
such anti-Rh(D) alloantibodies in a transfusion recipient who does
not naturally harbor D category III red blood cells. Although
providing transfused blood comprising D category III red blood
cells to a patient who does not naturally harbor such cells will
not necessarily cause immediate harm to the patient, the patient
thereby becomes alloimmunized against D category III red blood
cells. Such alloimmunized individuals may develop complications
including hemolytic transfusion reactions or hemolytic disease of
the newborn. TABLE-US-00146 TABLE 4 Analysis of Anti-RH(D) Clones
Obtained by Chain Shuffling. HEAVY CHAIN LIGHT CHAIN AGGLUTINATION
PATTERN.dagger-dbl. Rh(D) CLONE SEQUENCE.dagger. SEQUENCE.dagger.
wt III IVa IVb V VI VII SPECIFICITY SH04 SEQ ID NOs: 24/93 SEQ ID
NOs: 35/104 + + + + + 0 + epD 6/7 SH08 SEQ ID NOs: 12/81 SEQ ID
NOs: 154/197 + + + + + 0 + epD 6/7 SH10 SEQ ID NOs: 139/182 SEQ ID
NOs: 47/116 + 0 0 0 0 0 0 novel SH12 SEQ ID NOs: 9/78 SEQ ID NOs:
155/198 + + + + + 0 + epD 6/7 SH13 SEQ ID NOs: 26/95 SEQ ID NOs:
156/199 + 0 0 0 0 0 0 novel SH14 SEQ ID NOs: 24/93 SEQ ID NOs:
157/200 + + + + + 0 + epD 6/7 SH16 SEQ ID NOs: 140/183 SEQ ID NOs:
158/201 + 0 + + 0 0 0 novel SH17 SEQ ID NOs: 141/184 SEQ ID NOs:
47/116 + + 0 0 0 0 + epD 1 SH18 SEQ ID NOs: 142/185 SEQ ID NOs:
159/202 + 0 + + 0 0 0 novel SH20 SEQ ID NOs: 143/186 SEQ ID NOs:
160/203 + 0 + + + 0 0 novel SH21 SEQ ID NOs: 9/78 SEQ ID NOs:
161/204 + + + 0 + 0 0 novel SH24 SEQ ID NOs: 144/187 SEQ ID NOs:
162/205 + 0 0 0 0 0 0 novel SH25 SEQ ID NOs: 145/188 SEQ ID NOs:
35/104 + + 0 0 + 0 + epD 2 SH26 SEQ ID NOs: 21/90 SEQ ID NOs:
163/206 + + + 0 0 0 0 novel SH28 SEQ ID NOs: 146/189 SEQ ID NOs:
164/207 + + 0 0 + 0 + epD 2 SH30 SEQ ID NOs: 12/81 SEQ ID NOs:
165/208 + + + + + 0 + epD 6/7 SH32 SEQ ID NOs: 147/190 SEQ ID NOs:
166/209 + 0 0 0 0 0 0 novel SH34 SEQ ID NOs: 5/74 SEQ ID NOs:
167/210 + + 0 0 0 0 + epD 1 SH36 SEQ ID NOs: 14/83 SEQ ID NOs:
168/211 + 0 0 0 0 0 0 novel SH37 SEQ ID NOs: 148/191 SEQ ID NOs:
50/119 + + + 0 0 0 + epD X.sctn. SH39 SEQ ID NOs: 149/192 SEQ ID
NOs: 169/212 + 0 0 0 0 0 0 novel SH41 SEQ ID NOs: 24/93 SEQ ID NOs:
170/213 + + + + + 0 + epD 6/7 SH44 SEQ ID NOs: 150/193 SEQ ID NOs:
171/214 w* not determined SH46 SEQ ID NOs: 13/82 SEQ ID NOs:
172/215 + 0 + + 0 0 0 novel SH47 SEQ ID NOs: 151/194 SEQ ID NOs:
173/216 + + 0 0 + 0 + epD 2 SH48 SEQ ID NOs: 6/75 SEQ ID NOs:
174/217 + + 0 0 0 0 + epD 1 SH49 SEQ ID NOs: 17/86 SEQ ID NOs:
175/218 + + 0 0 0 0 + epD 1 SH50 SEQ ID NOs: 146/189 SEQ ID NOs:
176/219 + + 0 0 0 0 + epD 1 SH51 SEQ ID NOs: 17/86 SEQ ID NOs:
177/220 + + 0 0 + 0 + epD 2 SH52 SEQ ID NOs: 24/93 SEQ ID NOs:
178/221 + + 0 0 0 0 + epD 1 SH53 SEQ ID NOs: 146/189 SEQ ID NOs:
47/116 + 0 0 0 0 0 0 novel SH54 SEQ ID NOs: 152/195 SEQ ID NOs:
179/222 + + 0 0 0 0 + epD 1 SH55 SEQ ID NOs: 21/90 SEQ ID NOs:
180/223 + + 0 0 + 0 + epD 2 SH56 SEQ ID NOs: 153/196 SEQ ID NOs:
181/224 + + 0 0 0 0 0 novel Notes for TABLE 4 .dagger."SEQ ID NOs:
A/B" means that the chain had amino acid sequence "A" and was
encoded by nucleotide sequence "B". .dagger-dbl."+" means
agglutination occurred; "0" means agglutination did not occur.
*weak .sctn.as discussed in Example 2.
Amino Acid Sequences of Anti-Rh(D) Heavy and Light Chains
[0367] The amino acid sequences of various anti-Rh(D) antibody
chains were as follows, and are represented using single letter
amino acid codes.
[0368] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH10 is TABLE-US-00147
EVQLLEESGGGVVQPGRSLRLSCAASGFTFSRNG (SEQ ID NO:139)
MHWVRQAPGKGLEWVAFIWFDGSNKYYADSVKGR
FTISRDNSKNTLYLQMNSLRADDTAVYYCAREEA LFRGLTRWSYGMDVWGQGTTVSVSS.
[0369] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH16 is TABLE-US-00148
EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGM (SEQ ID NO:140)
HWVRQAPGRGLEWVALIWYDGGNKEYADSVKGRF
SISRDNSKNTLYLQVNSLRADDTAVYYCARDQRA AAGIFYYSRMDVWGQGTTVTVSS.
[0370] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH17 is TABLE-US-00149
EVQLLESGGGLVQPGGSLRLSCGASGIPFVSSWM (SEQ ID NO:141)
AWVRQAPGKGLEWVANIKQDGSKKNYVDSVEGRF
TISRDNAKNSLYLQMDSLRAEDTRIYYCARDSLT CFDYWGQGALVTVSS.
[0371] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH18 is TABLE-US-00150
EVQLLESGGGVVQPGRSLRLSCAASGFTFRSYAM (SEQ ID NO:142)
HWVRQAPGKGLEWVAATAYDGKNKYYADSVKGRF
TISRDNSMNTLFLQMNSLRAEDTAVFYCARGGFY YDSSGYYGLRHYFDSWGQGTLVTVSS.
[0372] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH20 is TABLE-US-00151
EVQLLEESGGGVVQPGRSLRLSCAASGFTFRSYA (SEQ ID NO:143)
MHWVRQAPGKGLEWVAVISYDGSTIYYADSVKGR
FTISRANSKNTLFLQMNSLRTEDTAVYYCTRGGF YYDSSGYYGLRHYFDYWGQGTLVTVSS.
[0373] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH24 is TABLE-US-00152
EVQLLESGGGVAQPGRSLRLSCVASGFSLRSYGM (SEQ ID NO:144)
HWVRQAPGKGLEWVADIWFDGSNKDYADSVKGRF
TISRDNSKNTLYLQMNSLRAEDTAVYYCARDWRV RAFSSGWLSAFDIWGQGTMVTVSS.
[0374] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH25 TABLE-US-00153
EVQLLEESGGGVVQPGRSLRLACAASGFSFRSYG (SEQ ID NO:145)
MHWVRQAPGRGLEWVAFTWFDGSNKYYVDSVKGR
FTISRDNSKNTLYLEMNSLRVDDTAVYYCAREAP MLRGISRYYYAMDVWGPGTTVTVSS.
[0375] The amino acid sequence of the heavy chain of each of
anti-Rh(D) antibody clones SH28, SH50, and SH53 is TABLE-US-00154
EVQLLESGGGGVQPGRSLRLSCAASGFTFNSYAM (SEQ ID NO:146)
YWVRQPPGKGLEWVAAIWYDGSNKEYADFVKGRF
TISRDNSKNTLSLQMNSLRDEDTAVYYCAREANL LRGWSRYYYGMDVWGQGTTVTVSS.
[0376] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH32 is TABLE-US-00155
EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGM (SEQ ID NO:147)
HWVRQAPGKGLEWVAFIWFDGSKNYYEDSVKGRF
TVSRDNSKNTLYLQMNSLRAEDTAVYYCARELSK KVALSRYYYYMDVWGQGTTVTVSS.
[0377] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH37 is TABLE-US-00156
EVQLLESGGGVVQPGRSLRLSCEASKFTLYNYGM (SEQ ID NO:148)
HWVRQAPGKGLEWVAFIWFDGSNKYYEDSVKGRF
TVSRDNSKNTLYLQMNSLRAEDTAVYYCARELSK KVALSRYYYYMDVWGQGTTVTVSS.
[0378] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH39 is TABLE-US-00157
EVQLLEQSGGGVVQPGRSLRLSCAASGFTFSSYG (SEQ ID NO:149)
MHWVRQAPGKGLEWVAVIWFDGSNKEYADSVKGR
FTISRDNSKNTLYLQMNSLRAEDTAVYYCAREEV VRGVILWSRKFDYWGQGTLVTVSS.
[0379] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH44 is TABLE-US-00158
EVQLLESGGGVAQPGRSLRLSCVASGFSLRSYGM (SEQ ID NO:150)
HWVRQAPGKGLEWVADIWFDGSNKDYADSVKGRF
TISRDNSKNTLYLQMNSLRAEDTAVYYCARDWRV RAFSSGWLSAFDIWGQGTMVTVSS.
[0380] The amino acid sequence of the heavy chain of anti-Rh(D))
antibody clone SH47 is TABLE-US-00159
EVQLLESGGGVVQPGRSLRLSCAASGFSFSNYAM (SEQ ID NO:151)
HWVRQAPGKGLEWVAVTSFDGSIKDYADSVKGRF
TISRDNSKNTLYLQMNSLRDEDTAVYYCARERGM IVVVRRRNAFDIWGQGTMVTVSS.
[0381] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH54 is TABLE-US-00160
EVQLLESGGGVVQPGRSLRLSCAASGFTFSRNGM (SEQ ID NO:152)
HWVRQAPGKGLEWVAFIWFDGSNKYYADSVKGRF
TISRDNSKNTLYLQMNSLRADDTAVYYCAREEAL FRGLTRWSYGMDVWGQGTTVSVSS.
[0382] The amino acid sequence of the heavy chain of anti-Rh(D)
antibody clone SH56 is TABLE-US-00161
EVQLLESGGGVVQPGRSLRLSCAASGFTFSSYGM (SEQ ID NO:153)
HWVRQAPGKGLEWVAVVYYDGSNKHYSDSVKGRF
TIFRDNSKNTLYLQMSLRAEDTAVYYCARERNFR SGYSRYYYGMDVWGPGTTVTVSS.
[0383] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH8 is TABLE-US-00162
AELTQSPSSLAASVGDRVTITCRANQTIRTSLNW (SEQ ID NO:154)
YQQRPGKAPNLLIYGASRLHSGVPSRFSGGISGA
DFTLTISSLQPEDFATYYCQQTYGYSRTFGQGTK VDIKR.
[0384] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH12 is TABLE-US-00163
AELTQSPFSLSASVGDRVTITCRASHNIYRSLNW (SEQ ID NO:155)
FQHKPGEAPKLLVYAASSLQRGVPTRFSGSGSGT
DFTLTISSLQPEDSATYFCQQSVTFPYTFGQGTK LEIRR.
[0385] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH13 is TABLE-US-00164
AELTQSPSSLSASVGDRVTITCRASQSISSYLNW (SEQ ID NO:156)
YQQKPGKAPKLLIYAASSLRSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSTPYTFGQGTK LEIKR.
[0386] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH14 is TABLE-US-00165
AELTQSPSSLSASVGDRVTITCRASQNIRRSLNW (SEQ ID NO:157)
YQHKPGRAPRLLIYAASTLQSGVPSRFRGSGSGT
DFTLTINSLQPADFATYYCQQSSNTPWTFGHGTK VEIKR.
[0387] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH16 is TABLE-US-00166
AELTQSPSSLSASVGDRVTITCRASQSISSYLNW (SEQ ID NO:158)
YQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSTPPTFGGGTK VEIKR.
[0388] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH18 is TABLE-US-00167
AELTQSPSSLSASVGDRVTITCRASQSISIALNW (SEQ ID NO:159)
YQQRPGKAPKLLMYATSTLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQYYNKPTFGPGTKV DIKR.
[0389] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH20 is TABLE-US-00168
AELTQSPFSLSASVGDRVTITCRASQSISRSLNW (SEQ ID NO:160)
YQHKPGEAPKLLIYAASSLQRGVPPRFSGSGSGT
DFTLTISSLQPEDFATYFCQQSVRIPYSFGQGTK LEIKR.
[0390] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH21 is TABLE-US-00169
AELTQSPSFLSASVGDRVTITCRASQGIRSYLAW (SEQ ID NO:161)
YQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGT
EFTLTIASLQPDDFATYYCQQLNNYPPFTFGPGT KVDIKR.
[0391] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH24 is TABLE-US-00170
AELTQSPSSLSASVGDRVTITCRASQSISTYLNW (SEQ ID NO:162)
YQQRPGKAPNLLIYAASTLQRGVPSRFTGSGSGT
DFTLTISSLQPEDFATYYCQQSYTTLWTFGQGTK MEIRR.
[0392] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH26 is TABLE-US-00171
AELTQSPSSLSASVGDRVTITCRASQSISSYLNW (SEQ ID NO:163)
YQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSFRRYSFGQGT KLEIKR.
[0393] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH28 is TABLE-US-00172
AELTQSPSSLSASVGDRVTITCRADQNIRRSLNW (SEQ ID NO:164)
FQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSSSTPWTFGRGTK VEIKR.
[0394] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH30 is TABLE-US-00173
YQQSPGKTPKLLIYAASSLQSGVPSRFSGSGSGT (SEQ ID NO:165)
DFTLTISSLQPEDFATYYCQQSYSTLTFGGGTKV EIKR.
[0395] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH32 is TABLE-US-00174
AELTQEPSLTVSPGGTVTLTCASSTGAVTSRYFP (SEQ ID NO:166)
NWFQQKPGQAPRALIYGSNNKHSWTPARFSGSLL
GGKAALTLSGVQPEDEAEYYCLLFYAGAWAFGGG TKLTVL.
[0396] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH34 is TABLE-US-00175
AELTQSPSSLSASVGDRVTITCRASQSISSYLNW (SEQ ID NO:167)
YQQKPGKAPKLLIYAASGLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSTPPYTFGQGT KLEIKR.
[0397] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH36 is TABLE-US-00176
AELTQSPSSLSASVGDRVTITCRASQSISSYLNW (SEQ ID NO:168)
YQQKPGKSPKLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSTPPAFGPGTK VDIKR.
[0398] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH39 is TABLE-US-00177
AELTQSPSSLSASVGDRVTITCRASQTIGRYLNW (SEQ ID NO:169)
YQQRPGKAPKLLVYAVSSLQSGAPSRFSGSGSGT
HFTLTITSLQPEDFATYCQQSYSSPFTFGQGTKV EIKR.
[0399] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH41 is TABLE-US-00178
AELTQSPSSLSASVGDRVTITCRASQNIRRSLNW (SEQ ID NO:170)
YQHKPGRAPRLLIYAASTLQSGVPSRFRGSGSGT
DFTLTINSLQPADFATYYCQQSSNTPWTFGHGTK VEIKR.
[0400] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH44 is TABLE-US-00179
AELTQSPSSLSASVGDRVIITCRASQTIPRFLNW (SEQ ID NO:171)
YQQKPGKAPVLLIHSISSLQSGVPSRFSASGSGT
EFTLTISSLQPEDFATYYCQQSYSNLSFGPGTTV DIRR.
[0401] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH46 is TABLE-US-00180
AELTQSPSSLSASVGDRVTITCRASQYISSYLNW (SEQ ID NO:172)
YQQKPGKAPNLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQTYSSPSTFGPGTK VDIKR.
[0402] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH47 is TABLE-US-00181
AELTQSPSSLSASVGDRVTITCRASQSISNYLNW (SEQ ID NO:173)
YQQKPGKAPNLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSYPRTFGQGQGTK VEIRR.
[0403] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH48 is TABLE-US-00182
AELTQSPSSLSASVGDRVTITCRASQYISSYLNW (SEQ ID NO:174)
YQQKPGKAPNLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQTYSSPSTFGPGTK VDIKR.
[0404] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH49 is TABLE-US-00183
AELTQSPSSLSASVGDRVTVTCRASQSISSYLNW (SEQ ID NO:175)
YQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSTPWFTGQGTK VEIKR.
[0405] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH50 is TABLE-US-00184
AELTQSPSSLSASVGDRVTVTCRASQSISSYLNW (SEQ ID NO:176)
YQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSTPWTFGQGTK VEIKR.
[0406] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH51 is TABLE-US-00185
AELTQSPSFLSASVGDRVTITCRASQGIRSYLAW (SEQ ID NO:177)
YQQKPGKAPKLLIYAASTLQSGVPSRFSGSGSGT
EFTLTISSLQPEDFATYYCQQLNNYPPFTFGPGT KVDIKR.
[0407] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH52 is TABLE-US-00186
AELTQSPGTLSLSPGERATLSCRASQSISSSYLA (SEQ ID NO:178)
WYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSG
TDFTLTISRLEPEDFAVYYCQQYGSSPWTFGQGT KVEIKR.
[0408] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH54 is TABLE-US-00187
AELTQSPSSMSASVGDRVTITCRASQSIGTYLNW (SEQ ID NO:179)
YQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQQSYSTPWTFGQGTK VEIKR.
[0409] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH55 is TABLE-US-00188
AELTQPPSASGTPGQRVTISCSGSSSNIGSKYVY (SEQ ID NO:180)
WYQQLPGTAPKLLIYSNNQRPSGVPDRFSAFKSG
TSASLAITGLQAEDEANYYCQSYDSGLSGWVFGG GTKLTVL.
[0410] The amino acid sequence of the light chain of anti-Rh(D)
antibody clone SH56 is TABLE-US-00189
AELTQSPSSLSASVGDRVTITCRASQSISRYLNW (SEQ ID NO:181)
YQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSGT
DFALTISSLLPEDFATYYCQQGYSTPPYSFGQGT KLEIKR.
Nucleotide Sequences of Anti-Rh(D) Heavy and Light Chains
[0411] The nucleotide sequences encoding various anti-Rh(D)
antibody clone chains were as follows.
[0412] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH10 is TABLE-US-00190
GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCG (SEQ ID NO:182)
TGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTG
TGCAGCGTCTGGGTTCACCTTCAGTAGGAATGGC
ATGCACTGGGTCCGCCAGGCTCCTGGCAAGGGGC
TGGAGTGGGGTGGCATTTATATGGTTTGATGGAA
GTAATAAATACTATGCAGACTCCGTGAAGGGCCG
ATTCACCATCTCCAGAGACAATTCCAAGAACACG
CTGTATCTGCAAATGAACAGCCTGAGAGCCGACG
ACACGGCTGTGTATTACTGTGCGAGAGAGGAGGC
TCTGTTTCGGGGACTTACTCGGTGGTCCTACGGC
ATGGACGTCTGGGGCCAAGGGACCACGGTCAGCG TCTCCTCA.
[0413] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH16 is TABLE-US-00191
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGG (SEQ ID NO:183)
TCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGC
AGCGTCTGGGTTCACCTTCAGTAGCTATGGCATG
CACTGGGTCCGCCAGGCTCCAGGCAGGGGGCTGG
AGTGGGTGGCTCTTATATGGTACGATGGAGGTAA
CAAAGAGTATGCAGACTCCGTGAAGGGCCGCTTC
AGCATCTCCAGAGACAACTCCAAGAACACTCTGT
ATCTGCAAGTGAACAGCCTGAGAGCCGACGACAC
GGCTGTCTATTACTGTGCGAGAGACCAGAGAGCA
GCAGCGGGTATCTTTTATTATTCCCGTATGGACG
TCTGGGGCCAAGGGACCACGGTCACCGTCTCCTC A.
[0414] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH17 is TABLE-US-00192
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCTTGG (SEQ ID NO:184)
TCCAGCCGGGGGGGTCCCTGAGACTCTCCTGTGG
TGCCTCTGGAATCCCCTTTGTTTCCTCTTGGATG
GCCTGGGTCCGCCAGGCCCCAGGGAAGGGGCTGG
AGTGGGTGGCCAACATAAAACAAGATGGAAGTAA
GAAAAACTATGTGGACTCTGTGGAGGGCCGATTC
ACCATCTCCAGAGACAACGCGAAGAACTCACTTT
ATCTGCAAATGGACAGCCTGAGAGCCGAGGACAC
GCGGATATATTACTGTGCGCGAGATTCACTTACT
TGTTTTGACTACTGGGGCCAGGGAGCCCTGGTCA CCGTCTCCTCA.
[0415] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH18 is TABLE-US-00193
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGG (SEQ ID NO:185)
TCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGC
AGCCTCTGGATTCACCTTCAGGAGCTATGCTATG
CACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG
AGTGGGTGGCAGCTACAGCATATGATGGAAAAAA
TAAATACTACGCAGACTCCGTGAAGGGCCGATTC
ACCATCTCCAGAGACAATTCCATGAACACGCTGT
TTCTGCAAATGAACAGCCTGAGAGCTGAGGACAC
GGCTGTGTTTTACTGTGCGAGAGGCGGATTTTAC
TATGATAGTAGTGGTTATTACGGCTTGAGGCACT
ACTTTGACTCCTGGGGCCAGGGAACCCTGGTCAC CGTCTCCTCA.
[0416] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH20 is TABLE-US-00194
GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCG (SEQ ID NO:186)
TGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTG
TGCAGCCTCTGGATTCACCTTCAGAAGTTATGCT
ATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGC
TGGAGTGGGTGGCGGTTATATCATATGATGGAAG
TACTATATACTACGCAGACTCCGTGAAGGGCCGA
TTCACCATCTCCAGAGCCAATTCCAAGAACACGC
TGTTTCTGCAAATGAACAGCCTCAGAACTGAGGA
CACGGCTGTATATTACTGTACGAGAGGGGGGTTT
TACTATGACAGTAGTGGTTATTACGGGTTGAGGC
ACTACTTTGACTACTGGGGCCAGGGAACCCTGGT CACCGTCTCTTCA.
[0417] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH24 is TABLE-US-00195
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGG (SEQ ID NO:187)
CCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGT
AGCGTCTGGATTCAGCCTCAGGAGCTATGGCATG
CACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGG
AGTGGGTGGCAGATATATGGTTTGATGGAAGTAA
TAAAGATTATGCAGACTCCGTGAAGGGCCGATTC
ACCATCTCCAGAGACAATTCCAAGAACACGTTGT
ATCTTCAAATGAACAGCCTGAGAGCCGAGGACAC
GGCTGTGTATTATTGTGCGAGAGATTGGAGGGTG
CGGGCCTTTAGTAGTGGCTGGTTAAGTGCTTTTG
ATATCTGGGGCCAAGGGACAATGGTCACCGTCTC TTCA.
[0418] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH25 is TABLE-US-00196
GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCG (SEQ ID NO:188)
TGGTCCAGCCTGGGAGGTCCCTGAGACTCGCCTG
TGCAGCGTCTGGATTCAGCTTCAGGAGCTATGGC
ATGCACTGGGTCCGCCAGGCTCCAGGCAGGGGGC
TGGAGTGGGTGGCATTTACATGGTTTGATGGAAG
CAATAAATATTATGTAGACTCCGTGAAGGGCCGA
TTCACCATCTCCAGAGACAATTCCAAGAACACGC
TGTATCTGGAAATGAACAGCCTGAGAGTCGATGA
CACGGTGGTATATTACTGTGCGAGAGAGGCGCCT
ATGCTTCGCGGAATTAGCAGATACTACTACGCGA
TGGACGTCTGGGGCCCAGGGACCACGGTCACCGT CTCCTCA.
[0419] The nucleotide sequence encoding the heavy chain of each of
anti-Rh(D) antibody clones SH28, SH50, and SH53 is TABLE-US-00197
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGGGG (SEQ ID NO:189)
TCCAGCCTGGGAGGTCCCTGCGACTCTCCTGTGC
GGCGTCTGGATTCACCTTCAATAGTTATGCCATG
TACTGGGTCCGCCAGCCTCCAGGCAAGGGGCTGG
AGTGGGTGGCAGCTATATGGTATGATGGAAGTAA
TAAAGAATATGCAGATTTTGTGAAGGGCCGCTTC
ACCATCTCCAGAGACAATTCCAAGAACACGCTGT
CTCTGCAAATGAACAGCCTGAGAGACGAGGACAC
GGCTGTGTATTACTGTGCGAGAGAGGCGAATCTC
CTCCGTGGCTGGTCTCGATACTACTACGGTATGG
ACGTCTGGGGCCAAGGGACCACGGTCACCGTCTC CTCA.
[0420] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH32 is TABLE-US-00198
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGG (SEQ ID NO:190)
TCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGA
AGCGTCTAAATTCACCCTCTACAATTATGGCATG
CACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG
AGTGGGTGGCATTTATATGGTTGATGGAAGTAAT
AAATACTATGAAGACTCCGTGAAGGGCCGATTCA
CCGTCTCCAGAGACAATTCCAAGAACACGCTGTA
TCTGCAAATGAACAGCCTGAGAGCCGAGGACACG
GCTGTGTATTACTGTGCGAGAGAACTATCTAAGA
AGGTGGCACTTTCTAGGTATTACTACTATATGGA
CGTCTGGGGCCAGGGGACCACGGTCACTGTCTCG TCA.
[0421] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH37 is TABLE-US-00199
GAGGTGCAGCTGCTCGAGGAGTCTGGGGGAGGCG (SEQ ID NO:191)
TGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTG
TGCAGTGTCTGGATTCACCCTAACTAATTATGGC
ATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGC
TGGAGTGGGTGGCACATGTCTGGTATGATGGAAG
TAAAACAGAATACGCAGACTCCGTCAAGGGCCGA
TTCGCCGTCTCCAGAGACAAATCCAAGAACACAC
TGTTTCTGCAAATGAACAGCCTGACAGCCGAGGA
CACGGCTATTTATTACTGTGCGAGAGAGAGGAGA
GAGAAAGTCTATATATTGTTCTACTCGTGGCTCG
ACCGCTGGGGCCAGGGAACCCTGGTCACCGTCTC CTCA.
[0422] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH39 is TABLE-US-00200
GAGGTGCAGCTGCTCGAGCAGTCTGGGGGAGGCG (SEQ ID NO:192)
TGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTG
TGCAGCGTCTGGATTCACCTTCAGTAGCTATGGC
ATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGAC
TGGAGTGGGTGGCAGTTATATGGTTTGATGGAAG
TAATAAGGAATATGCAGACTCCGTGAAGGGCCGA
TTCACCATCTCCAGAGACAATTCCAAGAACACGC
TGTATCTACAAATGAACAGCCTGAGAGCCGAGGA
CACGGCTGTGTATTACTGTGCGAGAGAAGAAGTG
GTTCGGGGAGTTATCTTATGGTCTCGGAAGTTTG
ACTACTGGGGCCAGGGAACCCTGGTCACCGTCTC CTCA.
[0423] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH44 is TABLE-US-00201
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGG (SEQ ID NO:193)
CCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGT
AGCGTCTGGATTCAGCCTCAGGAGCTATGGCATG
CACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGG
AGTGGGTGGCAGATATATGGTTTGATGGAAGTAA
TAAAGATTATGCAGACTCCGTGAAGGGCCGATTC
ACCATCTCCAGAGACAATTCCAAGAACACGTTGT
ATCTTCAAATGAACAGCCTGAGAGCCGAGGATAC
GGCTGTGTATTATTGTGCGAGAGATTGGAGGGTG
CGGGCCTTTAGTAGTGGCTGGTTAAGTGCTTTTG
ATATCTGGGGCCAAGGGACAATGGTCACCGTCTC TTCA.
[0424] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH47 is TABLE-US-00202
GAGGTGCAGCTGCTCGAGTCTGGGGGAGGCGTGG (SEQ ID NO:194)
TCCAGCCTGGGAGGTCCCTGCGACTCTCTTGTGC
AGCCTCTGGATTCAGCTTCAGTAACTATGCTATG
CACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG
AGTGGGTGGCAGTTACATCATTTGATGGAAGCAT
TAAAGACTACGCAGACTCCGTGAAGGGCCGATTC
ACCATCTCCAGAGACAATTCCAAGAACACACTAT
ATCTGCAAATGAACAGCCTGAGAGATGAGGACAC
GGCTGTATATTACTGTGCGAGAGAGCGGGGGATG
ATAGTCGTGGTCCGTCGCAGAAATGCTTTTGATA
TTTGGGGCCAAGGGACAATGGTCACCGTCTCTTC A.
[0425] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH54 is TABLE-US-00203
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGG (SEQ ID NO:195)
TCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGC
AGCGTCTGGGTTCACCTTCAGTAGGAATGGCATG
CACTGGGTCCGCCAGGCTCCTGGCAAGGGGCTGG
AGTGGGTGGCATTTATATGGTTTGATGGAAGTAA
TAAATACTATGCAGACTCCGTGAAGGGCCGATTC
ACCATCTCCAGAGACAATTCCAAGAACACGCTGT
ATCTGCAAATGAACAGCCTGAGAGCCGACGACAC
GGCTGTGTATTACTGTGCGAGAGAGGAGGCTCTG
TTTCGGGGACTTACTCGGTGGTCCTACGGTATGG
ACGTCTGGGGCCAAGGGACCACGGTCAGCGTCTC CTCA.
[0426] The nucleotide sequence encoding the heavy chain of
anti-Rh(D) antibody clone SH56 is TABLE-US-00204
GAGGTGCAGCTGCTCGAGTCGGGGGGAGGCGTGG (SEQ ID NO:196)
TCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGC
AGCGTCTGGATTCACCTTCAGTAGCTATGGCATG
CACTGGGTCCGGCAGGCTCCAGGCAAGGGGCTGG
AGTGGGTGGCAGTTGTCTACTATGATGGAAGTAA
CAAACACTATTCAGACTCCGTGAAGGGCCGATTC
ACCATCTTCAGAGACAACTCCAAGAACACGCTGT
ATCTACAAATGGACAGCCTGAGAGCCGAGGACAC
GGCTGTGTATTACTGTGCGAGAGAAAGAAATTTT
CGGAGTGGTTATTCCCGCTACTACTACGGTATGG
ACGTCTGGGGCCCAGGGACCACGGTCACCGTCTC CTCA.
[0427] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH8 is TABLE-US-00205
GCCGAGCTCACCCAGTCTCCATCCTCCCTGGCTG (SEQ ID NO:197)
CGTCTGTCGGAGACAGAGTCACCATCACTTGCCG
GGCAAATCAGACCATCAGAACCTCTTTAAATTGG
TATCAACAAAGACCTGGGAAAGCCCCTAACCTCC
TGATCTATGGTGCATCCAGGTTGCATAGTGGGGT
CCCATCAAGGTTTAGTGGCGGTATTTCTGGGGCA
GACTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAGCAGACTTA
CGGTTATTCTCGAACGTTCGGCCAAGGGACCAAG GTGGATATCAAACGA.
[0428] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH12 is TABLE-US-00206
GCCGAGCTCACCCAGTCTCCATTCTCCCTGTCTG (SEQ ID NO:198)
CATCTGTAGGAGACAGAGTCACCATAACTTGCCG
GGCAAGTCACAACATTTACAGGTCTTTAAATTGG
TTTCAGCATAAACCAGGGGAAGCCCCTAAGCTCC
TGGTCTATGCTGCATCCAGTCTGCAGCGTGGGGT
CCCAACCAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTTCAACCTG
AAGACTCTGCGACTTACTTCTGTCAACAGAGTGT
CACATTCCCCTACACTTTTGGCCAGGGGACCAAG CTGGAGATCAGACGA.
[0429] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH13 is TABLE-US-00207
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:199)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCGAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCCTACACTTTTGGCCAGGGGACCAAG CTGGAGATCAAACGA.
[0430] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH14 is TABLE-US-00208
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:200)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAACCATTAGGAGGTCTTTAAATTG
GTATCAACACAAACCAGGGAGAGCCCCTAGACTC
CTGATCTATGCTGCATCCACTTTGCAAAGTGGGG
TCCCATCAAGGTTCAGGGGCAGTGGATCTGGGAC
AGATTTCACTCTCACCATCAACAGTCTGCAACCT
GCAGATTTTGCAACTTACTACTGTCAGCAGAGTT
CCAATACCCCGTGGACGTTCGGCCATGGGACCAA GGTGGAAATCAAACGA.
[0431] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH16 is TABLE-US-00209
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:201)
CCTCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAACAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCTCCAACTTTCGGCGGAGGGACCAAG GTGGAGATCAAACGA.
[0432] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH18 is TABLE-US-00210
GCCGAGCTCACCCAGTCTCCATCCTCCCTCTCTG (SEQ ID NO:202)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGTATTAGCATCGCTTTAAATTGG
TATCAGCAGAGACCAGGGAAAGCCCCTAAGCTCC
TGATGTATGCTACATCCACTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAATATTA
CAATAAACCTACTTTCGGCCCTGGGACCAAGGTG GATATCAAACGA.
[0433] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH20 is TABLE-US-00211
GCCGAGCTCACCCAGTCTCCATTCTCCCTGTCTG (SEQ ID NO:203)
CATCTGTCGGAGACAGAGTCACCATAACTTGCCG
GGCAAGTCAGAGCATTAGCAGGTCTTTAAATTGG
TATCAACATAAACCAGGGGAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTCTGCAGCGTGGGGT
CCCACCCAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGACTTTGCGACTTACTTCTGTCAACAGAGTGT
CAGAATCCCGTACAGTTTGGCCAGGGGACCAAGC TGGAGATCAAACGA.
[0434] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH21 is TABLE-US-00212
GCCGAGCTCACCCAGTCTCCATCCTTCCTGTCTG (SEQ ID NO:204)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCCAGTCAGGGCATTAGGAGTTATTTAGCCTGG
TATCAGCAAAAACCAGGGAAAGCCCCTAAGCTCC
TAATCTATGCTGCATCCACTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGCGGCAGTGGATCTGGGACA
GAATTCACTCTCACAATCGCCAGCCTGCAGCCTG
ATGATTTTGCAACTTATTACTGTCAACAGCTTAA
TAATTACCCCCCTTTCACTTTCGGCCCTGGGACC AAAGTGGATATCAAACGA.
[0435] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH24 is TABLE-US-00213
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:205)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCACCTATTTAAATTGG
TATCAGCAGAGACCAGGGAAAGCCCCTAACCTCC
TGATCTATGCTGCATCCACTTTGCAAAGGGGGGT
CCCATCAAGGTTCACTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CACTACCCTGTGGACGTTCGGCCAAGGGACCAAG ATGGAAATCAGACGA.
[0436] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH26 is TABLE-US-00214
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:206)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGTTTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTTTCCGAAGGTACAGTTTTGGCCAGGGGACC AAGCTGGAGATCAAACGA.
[0437] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH28 is TABLE-US-00215
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:207)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAGATCAGAACATTAGGAGGTCTTTAAATTGG
TTTCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTC
CAGTACCCCGTGGACGTTCGGCCGAGGGACCAAG GTGGAAATCAAACGA.
[0438] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH30 is TABLE-US-00216
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:208)
CATCTGTTGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTCGGAGGTCTTTAAATTGG
TATCAGCAGAGTCCAGGGAAAACCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCTCACTTTCGGCGGAGGGACCAAGGTG GAGATCAAACGA.
[0439] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH32 is TABLE-US-00217
GCCGAGCTCACTCAGGAGCCCTCACTGACTGTGT (SEQ ID NO:209)
CCCCAGGAGGGACAGTCACTCTCACCTGTGCTTC
CAGCACTGGAGCAGTCACCAGTCGTTACTTTCCA
AACTGGTTCCAGCAGAAACCTGGCCAGGCACCCA
GGGCACTGATTTATGGTTCAAACAACAAACACTC
CTGGACCCCTGCCCGGTTCTCAGGCTCCCTCCTT
GGGGGCAAAGCTGCCCTGACACTGTCAGGTGTGC
AGCCTGAGGACGAGGCGGAGTATTACTGCCTGCT
CTTCTATGCTGGTGCTTGGGCGTTCGGCGGAGGG ACCAAGCTGACCGTCCTA.
[0440] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH34 is TABLE-US-00218
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:210)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCGGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCCCCGTACACTTTTGGCCAGGGGACC AAGCTGGAGATCAAACGA.
[0441] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH36 is TABLE-US-00219
GCCGAGCTCACTCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:211)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAATCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTCACTCTCACCATCAGCAGTCTGCAACCTGA
AGATTTTGCAACTTACTACTGTCAACAGAGTTAC
AGTACCCCTCCGGCTTTCGGCCCTGGGACCAAAG TGGATATCAAACGA.
[0442] The nucleotide sequence encoding the light chain of
anti-Rh(D)) antibody clone SH39 is TABLE-US-00220
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:212)
CATCTGTGGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGACCATTGGGAGGTATTTAAATTGG
TATCAGCAGAGGGCCAGGGAAAGCCCCCAAACTC
CTGGTATATGCTGTGTCCAGTTTGCAAAGTGGGG
CCCCATCAAGGTTCAGTGGCAGTGGCTCTGGGAC
ACATTTCACTCTCACCATCACCAGTCTGCAACCT
GAAGATTTTGCAACTTACTTCTGCCAACAGAGTT
ACAGTTCTCCTTTCACTTTTGGCCAGGGGACCAA GGTTGAGATCAAACGA.
[0443] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH41 is TABLE-US-00221
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:213)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAACATTAGGAGGTCTTTAAATTGG
TATCAACACAAACCAGGGAGAGCCCCTAGACTCC
TGATCTATGCTGCATCCACTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGGGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAACAGTCTGCAACCTG
CAGATTTTGCAACTTACTACTGTCAGCAGAGTTC
CAATACCCCGTGGACGTTCGGCCATGGGACCAAG GTGGAAATCAAACGA.
[0444] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH44 is TABLE-US-00222
GCCGAGCTCACCCAGTCTCCATCGTCCCTGTCTG (SEQ ID NO:214)
CATCTGTAGGAGACAGAGTCATCATCACTTGCCG
GGCAAGTCAGACCATTCCCAGGTTCTTGAATTGG
TATCAACAGAAGCCTGGAAAAGCCCCTGTTCTCC
TGATTCATAGTATATCCAGTTTACAAAGTGGGGT
CCCATCAAGGTTCAGTGCCAGTGGATCTGGGACA
GAGTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTCGCAACTTACTACTGCCAACAGAGTTA
CAGTAATCTCTCTTTCGGCCCTGGGACCACAGTG GATATTAGACGA.
[0445] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH46 is TABLE-US-00223
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:215)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGTACATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAATCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGACTTA
CAGTTCCCCTAGCACTTTCGGCCCTGGGACCAAA GTGGATATCAAACGA.
[0446] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH47 is TABLE-US-00224
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:216)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAACTATTTAAATTGG
TATCAGCAGAAACCAGGAAAAGCCCCTAACCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAAGGTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTTATCCTCGCACGTTCGGCCAAGGGACCAAG GTGGAGATCAGACGA.
[0447] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH48 is TABLE-US-00225
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:217)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGTACATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAATCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGACTTA
CAGTTCCCCTAGCACTTTCGGCCCTGGGACCAAG TGGATATCAAACGA.
[0448] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH49 is TABLE-US-00226
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:218)
CATCTGTAGGAGACAGAGTCACCGTCACTTGCCG
GGCAAGTCAGAGCATTAGCAGCTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCGTGGACGTTCGGCCAAGGGACCAAG GTGGAAATCAAACGA.
[0449] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH50 is TABLE-US-00227
GCCGAGCTCACCCAGTCTCCATCGTCCCTGTCTG (SEQ ID NO:219)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GACAAGTCAGAGCATTGGCACCTATTTAAATTGG
TATCAACAAAAACCAGGGAAAGCCCCTAAACTCC
TGATCTATGCTGCATCCAATGTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCGGTGGATCTGGGACA
GGTTTCTCTCTCATCATCAGCAGTCTGCAACCTG
AAGATTTAGCAATTTACTACTGCCAACAGAGCTA
CAGTGTCCCTCCGTACAGCTTTGGCCCGGGGACC AAGCTGGAGATCAAACGA.
[0450] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH51 is TABLE-US-00228
GCCGAGCTCACACAGTCTCCATCCTTCCTGTCTG (SEQ ID NO:220)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCCAGTCAGGGCATAAGGAGTTATTTAGCCTGG
TATCAGCAAAAACCAGGGAAAGCCCCTAAGCTCC
TAATCTATGCTGCATCCACTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGCGGCAGTGGATCTGGGACA
GAATTCACTCTCACAATCAGCAGCCTGCAGCCTG
AAGATTTTGCAACTTATTACTGTCAACAGCTTAA
TAATTACCCCCCTTTCACTTTCGGCCCTGGGACC AAAGTGGATATCAAACGA.
[0451] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH52 is TABLE-US-00229
GCCGAGCTCACACAGTCTCCAGGCACCCTGTCTT (SEQ ID NO:221)
TGTCTCCAGGGGAAAGAGCCACCCTCTCCTGCAG
GGCCAGTCAGAGTATTAGCAGCAGCTACTTAGCC
TGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGC
TCCTCATCTATGGTGCATCCAGCAGGGCCACTGG
CATCCCAGACAGATTCAGTGGCAGTGGGTCTGGG
ACAGACTTCACTCTCACCATCAGCAGACTGGAGC
CTGAAGATTTTGCAGTGTATTACTGTCAGCAGTA
TGGTAGCTCACCGTGGACGTTCGGCCAAGGGACC AAGGTGGAAATCAAACGA.
[0452] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH54 is TABLE-US-00230
GCCGAGCTCACCCAGTCTCCATCCTCCATGTCTG (SEQ ID NO:222)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTGGCACTTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCACTCTCACCATCAGCAGTCTGCAACCTG
AAGATTTTGCAACTTACTACTGTCAACAGAGTTA
CAGTACCCCGTGGACGTTCGGCCAAGGGACCAAG GTGGAAATCAAACGA.
[0453] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH55 is TABLE-US-00231
GCCGAGCTCACGCAGCCGCCCTCAGCGTCTGGGA (SEQ ID NO:223)
CCCCCGGGCAGAGGGTCACCATCTCTTGTTCTGG
AAGCAGCTCCAACATCGGAAGTAAATATGTATAC
TGGTACCAGCAACTCCCAGGAACGGCCCCCAAAC
TCCTCATTATAGTAATAATCAGCGGCCCTCAGGG
GTCCCTGACCGATTCTCTGCCTTCAAGTCTGGCA
CCTCAGCCTCCCTGGCCATCACTGGGCTCCAGGC
TGAGGATGAGGCTAATTATTACTGCCAGTCCTAT
GACAGCGGCCTGAGTGGCTGGGTGTTCGGCGGCG GGACCAAGCTGACCGTCCTA.
[0454] The nucleotide sequence encoding the light chain of
anti-Rh(D) antibody clone SH56 is TABLE-US-00232
GCCGAGCTCACCCAGTCTCCATCCTCCCTGTCTG (SEQ ID NO:224)
CATCTGTAGGAGACAGAGTCACCATCACTTGCCG
GGCAAGTCAGAGCATTAGCAGGTATTTAAATTGG
TATCAGCAGAAACCAGGGAAAGCCCCCAAGCTCC
TGATCTATGCTGCATCCAGTTTGCAAAGTGGGGT
CCCATCAAGGTTCAGTGGCAGTGGATCTGGGACA
GATTTCGCTCTCACCATCAGCAGTCTGCTACCTG
AAGATTTTGCAACTTACTACTGTCAACAGGGTTA
CAGTACCCCTCCGTACAGTTTTGGCCAGGGGACC AAGCTGGAGATCAAACGA.
[0455] The disclosures of each and every patent, patent application
and publication cited herein are hereby incorporated herein by
reference in their entirety.
[0456] While this invention has been disclosed with reference to
specific embodiments, it is apparent that other embodiments and
variations of this invention may be devised by others skilled in
the art without departing from the true spirit and scope of the
invention. The appended claims are intended to be construed to
include all such embodiments and equivalent variations.
Sequence CWU 1
1
224 1 128 PRT Homo sapiens anti-Rh(D) chain B01 1 Glu Val Gln Leu
Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr 20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35
40 45 Ala Ala Thr Ala Tyr Asp Gly Lys Asn Lys Tyr Tyr Ala Asp Ser
Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
Thr Leu Phe 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Phe Tyr Cys 85 90 95 Ala Arg Gly Gly Phe Tyr Tyr Asp Ser
Ser Gly Tyr Tyr Gly Leu Arg 100 105 110 His Tyr Phe Asp Ser Trp Gly
Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 2 124 PRT Homo
sapiens anti-Rh(D) chain C01 2 Glu Val Gln Leu Leu Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala
Ala Ser Gly Phe Ser Phe Ser Ser Tyr 20 25 30 Gly Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile
Ser Tyr Asp Gly His His Lys Asn Tyr Ala Asp Ser Val 50 55 60 Lys
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Lys Thr Leu Tyr 65 70
75 80 Leu Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr
Cys 85 90 95 Ala Asn Leu Arg Gly Glu Val Thr Arg Arg Ala Ser Val
Pro Phe Asp 100 105 110 Ile Trp Gly Pro Gly Thr Met Val Thr Val Ser
Ser 115 120 3 124 PRT Homo sapiens anti-Rh(D) chain C03 3 Glu Val
Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln His Gly Arg 1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Ser Tyr 20
25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45 Ser Val Ile Ser Tyr Asp Gly His His Lys Asn Tyr Ala
Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser
Lys Lys Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Pro Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Asn Leu Arg Gly Glu Val
Thr Arg Arg Ala Ser Val Pro Phe Asp 100 105 110 Ile Trp Gly Pro Gly
Thr Met Val Thr Val Ser Ser 115 120 4 124 PRT Homo sapiens
anti-Rh(D) chain C04 4 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Ser Phe Ser Thr Tyr 20 25 30 Gly Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Ser Tyr
Asp Gly His Asn Lys Asn Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Lys Thr Leu Tyr 65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Asn Leu Arg Gly Glu Val Thr Arg Arg Ala Ser Ile Pro Phe Asp
100 105 110 Ile Trp Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120
5 124 PRT Homo sapiens anti-Rh(D) chain C04 5 Glu Val Gln Leu Leu
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg
Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Ser Tyr 20 25 30 Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Ser Tyr Asp Gly Thr Asn Lys Tyr Phe Ala Asp Ser Val
50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Lys Thr
Leu Tyr 65 70 75 80 Leu Gln Met Thr Ser Leu Arg Pro Glu Asp Thr Ala
Val Tyr Phe Cys 85 90 95 Ala Asn Leu Arg Gly Glu Val Thr Arg Arg
Ala Ser Val Pro Leu Asp 100 105 110 Ile Trp Gly Gln Gly Thr Met Val
Thr Val Ser Ser 115 120 6 124 PRT Homo sapiens anti-Rh(D) chain C08
6 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1
5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Ser
Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
Glu Trp Val 35 40 45 Ala Val Ile Ser Tyr Asp Gly Thr Asn Lys Tyr
Phe Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Lys Thr Leu Tyr 65 70 75 80 Leu Gln Met Thr Ser Leu Arg
Pro Glu Asp Thr Ala Val Tyr Phe Cys 85 90 95 Ala Asn Leu Arg Gly
Glu Val Thr Arg Arg Ala Ser Val Pro Leu Asp 100 105 110 Ile Trp Gly
Gln Gly Thr Met Val Thr Val Ser Ser 115 120 7 124 PRT Homo sapiens
anti-Rh(D) chain C10 7 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Ser Phe Ser Ser Tyr 20 25 30 Gly Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Val Ile Ser Tyr
Asp Gly His His Lys Asn Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Lys Thr Leu Tyr 65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Pro Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Asn Leu Arg Gly Glu Val Thr Arg Arg Ala Ser Val Pro Phe Asp
100 105 110 Ile Trp Gly Pro Gly Thr Leu Val Thr Val Ser Ser 115 120
8 125 PRT Homo sapiens anti-Rh(D) chain D01 8 Glu Val Gln Leu Leu
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg
Leu Ser Cys Val Val Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30 Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asn Gln Ile Lys Leu Trp Ser
Arg Tyr Leu Tyr Tyr Phe 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120 125 9 125 PRT Homo sapiens anti-Rh(D)
chain D03 9 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro
Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Thr Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly
Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly Ser
Asn Lys Glu Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg
Glu Glu Val Val Arg Gly Val Ile Leu Trp Ser Arg Lys Phe 100 105 110
Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 10
126 PRT Homo sapiens anti-Rh(D) chain D04 10 Glu Val Gln Leu Leu
Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg
Leu Ser Cys Val Ala Ser Gly Phe Ser Leu Arg Ser Tyr 20 25 30 Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Asp Ile Trp Phe Asp Gly Ser Asn Lys Asp Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Trp Arg Val Arg Ala Phe Ser
Ser Gly Trp Leu Ser Ala 100 105 110 Phe Asp Ile Trp Gly Gln Gly Thr
Met Val Thr Val Ser Ser 115 120 125 11 127 PRT Homo sapiens
anti-Rh(D) chain D05 11 Glu Val Gln Leu Leu Glu Glu Ser Gly Gly Gly
Val Ala Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser Cys Val Ala
Ser Gly Phe Ser Leu Arg Ser 20 25 30 Tyr Gly Met His Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Ala Asp Ile Trp
Phe Asp Gly Ser Asn Lys Asp Tyr Ala Asp Ser 50 55 60 Val Lys Gly
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70 75 80 Tyr
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr 85 90
95 Cys Ala Arg Asp Trp Arg Val Arg Ala Phe Ser Ser Gly Trp Leu Ser
100 105 110 Ala Phe Asp Ile Trp Gly Gln Gly Thr Thr Val Ser Val Ser
Ser 115 120 125 12 125 PRT Homo sapiens anti-Rh(D) chain D07 12 Glu
Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10
15 Ser Leu Arg Leu Ser Cys Ala Val Ser Gly Phe Thr Leu Thr Asn Tyr
20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45 Ala His Val Trp Tyr Asp Gly Ser Lys Thr Glu Tyr
Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Ala Val Ser Arg Asp Lys
Ser Lys Asn Thr Leu Phe 65 70 75 80 Leu Gln Met Asn Ser Leu Thr Ala
Glu Asp Thr Ala Ile Tyr Tyr Cys 85 90 95 Ala Arg Glu Arg Arg Glu
Lys Val Tyr Ile Leu Phe Tyr Ser Trp Leu 100 105 110 Asp Arg Trp Gly
Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125 13 126 PRT Homo
sapiens anti-Rh(D) chain D08 13 Glu Val Gln Leu Leu Glu Glu Ser Gly
Gly Gly Val Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Ser Ser 20 25 30 Tyr Gly Met His Trp
Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp 35 40 45 Val Ala Leu
Ile Trp Tyr Asp Gly Gly Asn Lys Glu Tyr Ala Asp Ser 50 55 60 Val
Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70
75 80 Tyr Leu Gln Val Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr
Tyr 85 90 95 Cys Ala Arg Asp Gln Arg Ala Ala Ala Gly Ile Phe Tyr
Tyr Ser Arg 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 125 14 126 PRT Homo sapiens anti-Rh(D) chain
D09 14 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly
Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Lys Phe Thr Leu
Tyr Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Trp Phe Asp Gly Ser Asn
Lys Tyr Tyr Glu Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu
Gly Ser Lys Lys Val Ala Leu Ser Arg Tyr Tyr Tyr Tyr 100 105 110 Met
Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 15
126 PRT Homo sapiens anti-Rh(D) chain D10 15 Glu Val Gln Leu Leu
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg
Leu Ser Cys Glu Ala Ser Lys Phe Thr Leu Tyr Asn Tyr 20 25 30 Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Phe Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Glu Asp Ser Val
50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Gly Ser Lys Lys Val Ala Leu
Ser Arg Tyr Tyr Tyr Tyr 100 105 110 Met Asp Val Trp Gly Gln Gly Thr
Thr Val Thr Val Ser Ser 115 120 125 16 126 PRT Homo sapiens
anti-Rh(D) chain D11 16 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser
Lys Phe Thr Leu Tyr Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln
Ala Pro Gly Glu Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Trp Phe
Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg
Phe Thr Val Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Arg Glu Val Ser Lys Lys Leu Ala Leu Ser Arg Tyr Tyr Tyr Tyr
100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
115 120 125 17 126 PRT Homo sapiens anti-Rh(D) chain D12 17 Glu Val
Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15
Ser Leu Arg Leu Ala Cys Ala Ala Ser Gly Phe Ser Phe Arg Ser Tyr 20
25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp
Val 35 40 45 Ala Phe Thr Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Val
Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser
Lys Asn Thr Leu Tyr 65 70 75 80 Leu Glu Met Asn Ser Leu Arg Val Asp
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Ala Ser Met Leu
Arg Gly Ile Ser Arg Tyr Tyr Tyr Ala 100 105 110 Met Asp Val Trp Gly
Pro Gly Thr Thr Val Thr Val Ser Ser 115 120 125 18 127 PRT Homo
sapiens anti-Rh(D) chain D13 18 Glu Val Gln Leu Leu Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala
Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Gly Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile
Trp Phe Asp Gly Ser Asn Arg Asp Tyr Ala Glu Ser Val 50 55 60 Lys
Gly Arg Phe Thr Ile Ser Arg Asp Lys Ser Lys Asn Thr Leu Tyr 65 70
75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr
Cys 85 90 95 Ala Arg Glu Asn Val Ala Arg Gly Gly Gly Gly Val Arg
Tyr Lys Tyr 100 105 110 Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val
Thr Val Ser Ser 115 120 125 19 127 PRT Homo sapiens anti-Rh(D)
chain D14 19 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Ser Thr Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly
Ser Lys Arg Asp Tyr Ala Glu Ser Val 50 55 60 Lys Gly Arg Phe Thr
Ile Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu
Asn Val Ala Arg Gly Gly Gly Gly Ile Arg Tyr Lys Tyr 100 105 110 Tyr
Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125
20 125 PRT Homo sapiens anti-Rh(D) chain D15 20 Glu Val Gln Leu Leu
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg
Leu Ser Cys Val Val Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30 Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asn Gln Ile Lys Leu Trp Ser
Arg Tyr Leu Tyr Tyr Phe 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120 125 21 125 PRT Homo sapiens anti-Rh(D)
chain D16 21 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Val Ser Gly Phe
Thr Phe Asn Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly
Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Glu Asn Gln Ile Lys Leu Trp Ser Arg Tyr Leu Tyr Tyr Phe 100 105
110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125
22 125 PRT Homo sapiens anti-Rh(D) chain D17 22 Glu Val Gln Leu Leu
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg
Leu Ser Cys Val Val Ser Gly Phe Thr Phe Asn Asn Tyr 20 25 30 Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Asn Gln Ile Lys Leu Trp Ser
Arg Tyr Leu Tyr Tyr Phe 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120 125 23 125 PRT Homo sapiens anti-Rh(D)
chain D18 23 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Val Val Ser Gly Phe
Thr Phe Asn Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Ser
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Ile Trp Phe Asp Gly
Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Glu Asn Gln Ile Lys Leu Trp Ser Arg Tyr Leu Tyr Tyr Phe 100 105
110 Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120 125
24 125 PRT Homo sapiens anti-Rh(D) chain D20 24 Glu Val Gln Leu Leu
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30 Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Glu Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Glu Val Val Arg Gly Val Ile
Leu Trp Ser Arg Lys Phe 100 105 110 Asp Tyr Trp Gly Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120 125 25 126 PRT Homo sapiens anti-Rh(D)
chain D30 25 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Ser Ser Tyr 20 25 30 Gly Met Arg Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Val Val Tyr Tyr Asp Gly
Ser Asn Lys His Tyr Ser Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met
Asp Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala
Arg Glu Arg Asn Phe Arg Ser Gly Tyr Ser Arg Tyr Tyr Tyr Gly 100 105
110 Met Asp Val Trp Gly Pro Gly Thr Thr Val Thr Val Ser Ser 115 120
125 26 126 PRT Homo sapiens anti-Rh(D) chain D31 26 Glu Val Gln Leu
Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30
Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35
40 45 Ala Val Val Tyr Tyr Asp Gly Ser Asn Lys His Tyr Ser Asp Ser
Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
Thr Leu Tyr 65 70 75 80 Leu Gln Met Asp Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Arg Asn Phe Arg Ser Gly
Tyr Ser Arg Tyr Tyr Tyr Gly 100 105 110 Met Asp Val Trp Gly Pro Gly
Thr Thr Val Thr Val Ser Ser 115 120 125 27 127 PRT Homo sapiens
anti-Rh(D) chain E01is 27 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly
Leu Val Lys Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ser Met His Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ser Ile Ser
Asn Ser Asn Thr Tyr Ile Tyr Tyr Ala Asp Ala Val 50 55 60 Lys Gly
Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80
Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Asp Ser Arg Tyr Ser Asn Phe Leu Arg Trp Val Arg Ser
Asp 100 105 110 Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Ile Val
Ser Ser 115 120 125 28 131 PRT Homo sapiens anti-Rh(D) chain E03 28
Glu Val Gln Leu Leu Glu Ser Gly Val Glu Ser Gly Gly Gly Leu Val 1 5
10 15 Lys Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr 20 25 30 Phe Ser Ser Tyr Ser Met His Trp Val Arg Gln Gly Pro
Gly Lys Gly 35 40 45 Leu Glu Trp Val Ser Ser Ile Ser Asn Ser Asn
Thr Tyr Ile Tyr Tyr 50 55 60 Ala Asp Ala Val Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ala Lys 65 70 75 80 Asn Ser Leu Tyr Leu Gln Met
Asn Ser Leu Arg Ala Glu His Thr Ala 85 90 95 Val Tyr Tyr Cys Ala
Arg Asp Ser Arg Tyr Ser Asn Phe Leu Arg Trp 100 105 110 Val Arg Ser
Asp Gly Met Asp Val Trp Gly Gln Gly Thr Thr Val Ile 115 120 125 Val
Ser Ser 130 29 107 PRT Homo sapiens anti-Rh(D) chain F01 29 Ala Glu
Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15
Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Phe Arg Asn Asp Leu 20
25 30 Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile
Tyr 35 40 45 Ala Thr Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe
Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Asn
Ser Leu Gln Pro Glu 65 70 75 80 Asp Ser Ala Thr Tyr Tyr Cys Leu Gln
His Asn Ser Phe Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val
Glu Ile Lys Arg 100 105 30 112 PRT Homo sapiens anti-Rh(D) chain
G01 30 Ala Glu Leu Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly
Glu 1 5 10 15 Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu
His Ser Ser 20 25 30 Gly Phe Asn Phe Leu Asp Trp Tyr Leu Gln Lys
Pro Gly Gln Ser Pro 35 40 45 Gln Leu Leu Ile Tyr Met Gly Ser Asn
Arg Ala Ser Gly Val Pro Asp 50 55 60 Arg Phe Ser Gly Ser Gly Ser
Gly Thr Asp Phe Thr Leu Lys Ile Asn 65 70 75 80 Arg Val Glu Ala Glu
Asp Val Gly Val Tyr Tyr Cys Met Gln Ala Leu 85 90 95 Gln Phe Pro
Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100 105 110 31
108 PRT Homo sapiens anti-Rh(D) chain H01 31 Ala Glu Leu Thr Gln
Ser Pro Ser Phe Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Gly Ile Thr Ser Tyr Leu 20 25 30 Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40
45 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ala Ser Leu Gln
Pro Asp 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Asn
Tyr Pro Pro Phe 85 90 95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile
Lys Arg 100 105 32 108 PRT Homo sapiens anti-Rh(D) chain I01 32 Ala
Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10
15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu
20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg
Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln
Gln Ser Tyr Ser Thr Pro Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys Arg 100 105 33 107 PRT Homo sapiens anti-Rh(D)
chain I02 33 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser
Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser
Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys
Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser
Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala
Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Leu Trp Thr 85 90 95 Phe
Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 34 107 PRT Homo
sapiens anti-Rh(D) chain I03 34 Ala Glu Leu Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Ala Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg
Thr Ser Arg Asn Ile Asn Arg Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser
Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Thr Ser Leu Gln Pro Glu 65 70
75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Phe
Thr 85 90 95 Phe Gly Pro Gly Thr Lys Val Asp Leu Lys Arg 100 105 35
107 PRT Homo sapiens anti-Rh(D) chain I04 35 Ala Glu Leu Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Asn Ile Arg Arg Ser Leu 20 25 30 Asn
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40
45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn
Thr Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
Arg 100 105 36 107 PRT Homo sapiens anti-Rh(D) chain I05 36 Ala Glu
Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15
Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Arg Arg Tyr Leu 20
25 30 Asn Trp Tyr Gln His Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
Phe 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe
Thr Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
Ser Tyr Ser Thr Pro Gln Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val
Glu Ile Lys Arg 100 105 37 107 PRT Homo sapiens anti-Rh(D) chain
I06 37 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser
Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val
Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr
Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr
Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Ile Thr 85 90 95 Phe Gly Gln
Gly Thr Arg Leu Glu Ile Lys Arg 100 105 38 107 PRT Homo sapiens
anti-Rh(D) chain I07 38 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser
Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro
Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu
Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp
Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Arg Thr 85 90
95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100 105 39 107 PRT
Homo sapiens anti-Rh(D) chain I08 39 Ala Glu Leu Thr Gln Ser Pro
Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr
Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr
Gln
Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala
Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65
70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro
Arg Thr 85 90 95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100
105 40 107 PRT Homo sapiens anti-Rh(D) chain I09 40 Ala Glu Leu Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35
40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
Ser 50 55 60 Gly Ser Gly Thr Asp Ser Thr Leu Thr Ile Ser Ser Leu
Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn
Ser Tyr Pro Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg 100 105 41 108 PRT Homo sapiens anti-Rh(D) chain I10 41 Ala
Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10
15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Ser Ser Tyr Leu
20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Leu Ser Arg
Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln
Gln Ser Tyr Ser Thr Pro Pro Tyr 85 90 95 Ser Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys Arg 100 105 42 103 PRT Homo sapiens anti-Rh(D)
chain I11 42 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser
Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser
Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys
Ala Pro Thr Leu Leu Ile Asn 35 40 45 Ala Ala Ser Ser Leu Gln Ser
Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala
Ile Tyr Tyr Cys Gln Gln Arg Glu Thr Phe Gly Gln Gly 85 90 95 Thr
Lys Leu Glu Ile Lys Arg 100 43 108 PRT Homo sapiens anti-Rh(D)
chain I12 43 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser
Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser
Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys
Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser
Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala
Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro Tyr 85 90 95 Thr
Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 44 107 PRT Homo
sapiens anti-Rh(D) chain I13 44 Ala Glu Leu Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg
Ala Ser Gln Ser Ile Ser Arg Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser
Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70
75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Gly Thr Pro His
Ser 85 90 95 Phe Gly Arg Gly Thr Lys Leu Glu Ile Lys Arg 100 105 45
107 PRT Homo sapiens anti-Rh(D) chain I15 45 Ala Glu Leu Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr
Ile Thr Cys Arg Ala Asn Gln Asn Ile Arg Arg Ser Leu 20 25 30 Asn
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 40
45 Ala Ala Ser Thr Leu Gln Gly Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Leu Ala 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Ser Ala
Thr Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
Arg 100 105 46 107 PRT Homo sapiens anti-Rh(D) chain I16 46 Ala Glu
Leu Thr Gln Ser Pro Ser Ser Leu Pro Ala Ser Val Gly Asp 1 5 10 15
Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Thr Ile Gly Phe Asn Leu 20
25 30 Asn Trp Tyr Gln Gln Thr Ser Gly Lys Pro Pro Lys Leu Leu Ile
Tyr 35 40 45 Gly Val Ser Lys Leu Gln Asn Gly Val Pro Ser Arg Phe
Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
Thr Asn Asp Ala Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val
Glu Val Arg Arg 100 105 47 106 PRT Homo sapiens anti-Rh(D) chain
J01 47 Ala Glu Leu Gln Asp Pro Val Val Ser Val Ala Leu Gly Gln Thr
Val 1 5 10 15 Arg Ile Thr Cys Gln Gly Asp Gly Leu Arg Ser Tyr Tyr
Ala Ser Trp 20 25 30 Tyr Gln Gln Lys Pro Gly Gln Ala Pro Lys Leu
Val Met Tyr Gly Arg 35 40 45 Asn Asn Arg Pro Ser Gly Ile Pro Gly
Arg Phe Ser Gly Ser Ser Ser 50 55 60 Gly Gln Thr Ala Ala Leu Thr
Ile Thr Gly Thr Gln Ala Glu Asp Glu 65 70 75 80 Ala Asp Tyr Tyr Cys
Gln Ser Arg Ala Thr Ser Gly Asn Pro Val Val 85 90 95 Phe Gly Gly
Gly Thr Lys Leu Thr Val Leu 100 105 48 106 PRT Homo sapiens
anti-Rh(D) chain J02 48 Ala Glu Leu Gln Asp Pro Val Val Ser Val Ala
Leu Gly Gln Thr Val 1 5 10 15 Arg Ile Thr Cys Gln Gly Asp Gly Leu
Arg Ser Tyr Tyr Ala Ser Trp 20 25 30 Tyr Gln Gln Lys Pro Gly Gln
Ala Pro Lys Leu Val Met Tyr Gly Arg 35 40 45 Asn Asn Arg Pro Ser
Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser 50 55 60 Gly Gln Thr
Ala Ala Leu Thr Ile Thr Gly Thr Gln Ala Glu Asp Glu 65 70 75 80 Ala
Asp Tyr Tyr Cys Gln Ser Arg Ala Thr Ser Gly Asn Pro Val Val 85 90
95 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 49 104 PRT Homo
sapiens anti-Rh(D) chain J04 49 Ala Glu Leu Gln Asp Pro Val Val Ser
Val Ala Leu Gly Gln Thr Val 1 5 10 15 Arg Ile Thr Cys Gln Gly Asp
Ser Leu Arg Ser Tyr Tyr Ala Ser Trp 20 25 30 Tyr Gln Gln Lys Pro
Gly Gln Ala Pro Val Leu Val Ile Tyr Gly Lys 35 40 45 Asn Ser Arg
Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Ser Ser 50 55 60 Gly
Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu Asp Glu 65 70
75 80 Ala Asp Tyr Tyr Cys Ser Ser Arg Gly Ser Pro His Val Ala Phe
Gly 85 90 95 Gly Gly Thr Lys Leu Thr Val Leu 100 50 106 PRT Homo
sapiens anti-Rh(D) chain J05 50 Ala Glu Leu Gln Asp Pro Val Val Ser
Val Ala Leu Gly Gln Thr Val 1 5 10 15 Lys Ile Thr Cys Gln Gly Asp
Ser Leu Arg Lys Tyr Tyr Ala Ser Trp 20 25 30 Tyr Gln Gln Lys Pro
Gly Gln Ala Pro Val Leu Val Phe Tyr Ala Arg 35 40 45 Asn Ser Arg
Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser Asn Ser 50 55 60 Gly
Thr Thr Ala Ser Leu Thr Ile Ala Gly Ala Arg Ala Glu Asp Glu 65 70
75 80 Ala Asp Tyr Tyr Cys His Ser Arg Asp Ser Asn Gly His His Arg
Val 85 90 95 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 51 108
PRT Homo sapiens anti-Rh(D) chain K01 51 Ala Glu Leu Thr Gln Glu
Pro Ser Leu Thr Val Ser Pro Gly Gly Thr 1 5 10 15 Val Thr Leu Thr
Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Arg Tyr 20 25 30 Phe Pro
Asn Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro Leu 35 40 45
Ile Tyr Ser Ala Ser Asn Lys His Ser Trp Thr Pro Ala Arg Phe Ser 50
55 60 Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val
Gln 65 70 75 80 Pro Glu Asp Glu Ala Glu Tyr Tyr Cys Leu Leu Tyr Tyr
Ser Gly Ala 85 90 95 Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val
Leu 100 105 52 108 PRT Homo sapiens anti-Rh(D) chain K02 52 Ala Glu
Leu Thr Gln Glu Pro Ser Leu Thr Val Ser Pro Gly Gly Thr 1 5 10 15
Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala Val Thr Ser Arg Tyr 20
25 30 Phe Pro Asn Trp Phe Gln Gln Lys Pro Gly Gln Ala Pro Arg Pro
Leu 35 40 45 Ile Tyr Ser Ala Ser Asn Lys His Ser Trp Thr Pro Ala
Arg Phe Ser 50 55 60 Gly Ser Leu Leu Gly Gly Lys Ala Ala Leu Thr
Leu Ser Gly Val Gln 65 70 75 80 Pro Glu Asp Glu Ala Glu Tyr Tyr Cys
Leu Leu Tyr Tyr Ser Gly Ala 85 90 95 Trp Val Phe Gly Gly Gly Thr
Lys Leu Thr Val Leu 100 105 53 108 PRT Homo sapiens anti-Rh(D)
chain K03 53 Ala Glu Leu Thr Gln Pro Pro Ser Leu Thr Val Ser Pro
Gly Gly Thr 1 5 10 15 Val Thr Leu Thr Cys Ala Ser Ser Thr Gly Ala
Val Thr Ser Arg Tyr 20 25 30 Phe Pro Asn Trp Phe Gln Gln Lys Pro
Gly Gln Ala Pro Arg Ala Leu 35 40 45 Ile Tyr Gly Ser Asn Asn Lys
His Ser Trp Thr Pro Ala Arg Phe Ser 50 55 60 Gly Ser Leu Leu Gly
Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln 65 70 75 80 Pro Glu Asp
Glu Ala Glu Tyr Tyr Cys Leu Leu Phe Tyr Ala Gly Ala 85 90 95 Trp
Ala Phe Gly Gly Trp Thr Lys Leu Thr Val Leu 100 105 54 109 PRT Homo
sapiens anti-Rh(D) chain L01 54 Ala Glu Leu Thr Gln Pro Pro Ser Ala
Ser Gly Thr Pro Gly Gln Arg 1 5 10 15 Val Thr Ile Ser Cys Ser Gly
Gly Ser Ser Asn Ile Ala Ser Asn Thr 20 25 30 Val Asn Trp Tyr Gln
Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Asn
Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser
Lys Ser Gly Thr Ser Ala Thr Leu Val Ile Thr Gly Leu Gln Thr 65 70
75 80 Gly Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp His Ser Arg
Ser 85 90 95 Gly Ala Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 55 109 PRT Homo sapiens anti-Rh(D) chain L03 55 Ala Glu Leu
Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg 1 5 10 15 Val
Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Asn Asn His 20 25
30 Val Ser Trp Tyr Gln Gln Leu Pro Gly Met Ala Pro Lys Leu Leu Ile
35 40 45 Tyr Ser Asn Gly Gln Arg Pro Ser Gly Val Pro Asp Arg Phe
Ser Gly 50 55 60 Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser
Gly Leu Gln Ser 65 70 75 80 Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala
Trp His Asp Ser Leu Tyr 85 90 95 Gly Pro Val Phe Gly Gly Gly Thr
Lys Leu Thr Val Leu 100 105 56 109 PRT Homo sapiens anti-Rh(D)
chain L04 56 Ala Glu Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro
Gly Gln Arg 1 5 10 15 Val Ser Ile Ser Cys Ser Gly Ser Ser Ser Asn
Ile Gly Ser Asn Thr 20 25 30 Val Asn Trp Tyr Gln Gln Leu Pro Gly
Thr Ala Pro Lys Leu Leu Ile 35 40 45 Ser Thr Asn Asn Gln Gly Pro
Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Lys Ser Gly Thr
Ser Ser Ser Leu Ala Ile Ser Gly Leu Arg Ser 65 70 75 80 Glu Ala Glu
Asp Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Thr Leu Asn 85 90 95 Gly
Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 57 109 PRT
Homo sapiens anti-Rh(D) chain L05 57 Ala Glu Leu Thr Gln Pro Pro
Ser Ala Ser Gly Thr Pro Gly Leu Arg 1 5 10 15 Val Thr Ile Ser Cys
Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Ile 20 25 30 Val Asn Trp
Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 35 40 45 Phe
Ser Asn Asn Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55
60 Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln Ser
65 70 75 80 Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Thr Trp Asp Asp Ser
Leu Asn 85 90 95 Gly Arg Val Phe Gly Gly Gly Thr Lys Leu Thr Val
Leu 100 105 58 109 PRT Homo sapiens anti-Rh(D) chain M01 58 Ala Glu
Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln Arg 1 5 10 15
Val Thr Ile Ser Cys Ser Gly Ser Asn Phe Asn Ile Gly Ser Asn Tyr 20
25 30 Val Phe Trp Tyr Gln His Val Pro Gly Thr Ala Pro Lys Leu Leu
Ile 35 40 45 Tyr Asn Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg
Leu Ser Gly 50 55 60 Ser Lys Ser Gly Ala Ser Ala Ser Leu Ala Ile
Asn Gly Leu Arg Ser 65 70 75 80 Asp Asp Glu Ala Asp Tyr Tyr Cys Thr
Gly Trp Asp Asp Arg Leu Ser 85 90 95 Gly Leu Ile Phe Gly Gly Gly
Pro Lys Val Thr Val Leu 100 105 59 109 PRT Homo sapiens anti-Rh(D)
chain M02 59 Ala Glu Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro
Gly Gln Arg 1 5 10 15 Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn
Ile Gly Ser Asn Tyr 20 25 30 Val Tyr Trp Tyr Gln Gln Leu Pro Gly
Thr Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Arg Asn Asn Gln Arg Pro
Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55 60 Ser Lys Ser Gly Thr
Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg Ser 65 70 75 80 Glu Asp Glu
Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu Ser 85 90 95 Gly
Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 60 110 PRT
Homo sapiens anti-Rh(D) chain M03 60 Ala Glu Leu Thr Gln Pro Pro
Ser Ala Ser Gly Thr Pro Gly Gln Arg 1 5 10 15 Val Thr Ile Ser Cys
Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn Tyr 20 25 30 Val Tyr Trp
Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu Ile 35 40 45 Tyr
Arg Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser Gly 50 55
60 Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg Ser
65 70 75 80 Glu Ala Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser
Leu Ser 85 90 95 Ala Val Val Phe Gly Gly Gly Thr Lys Leu Thr
Val
Leu Leu 100 105 110 61 109 PRT Homo sapiens anti-Rh(D) chain N01 61
Ala Glu Leu Thr Gln Pro Pro Ser Val Ser Ala Ala Pro Gly Gln Lys 1 5
10 15 Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Asp Ser Asn
Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys
Leu Leu Ile 35 40 45 Phe Asp Asn Tyr Arg Arg Pro Ser Gly Ile Pro
Asp Arg Phe Ser Gly 50 55 60 Ser Lys Ser Gly Thr Ser Ala Thr Leu
Gly Ile Thr Gly Leu Gln Thr 65 70 75 80 Gly Asp Glu Ala Asp Tyr Tyr
Cys Ala Thr Trp Asp Asp Ser Leu Asn 85 90 95 Gly Arg Val Phe Gly
Gly Gly Thr Lys Leu Thr Val Leu 100 105 62 114 PRT Homo sapiens
anti-Rh(D) chain N02 62 Ala Glu Leu Thr Gln Pro Pro Ser Val Ser Ala
Ala Pro Gly Gln Lys 1 5 10 15 Val Thr Ile Ser Cys Ser Gly Ser Ser
Ser Asn Ile Gly Asn Asn Tyr 20 25 30 Val Ser Trp Tyr Gln Gln Leu
Pro Gly Thr Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Asn Asn Lys
Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly 50 55 60 Ser Lys Ser
Gly Thr Ser Ala Thr Leu Gly Ile Thr Gly Leu Gln Thr 65 70 75 80 Gly
Asp Glu Ala Asp Tyr Tyr Cys Gly Thr Trp Asp Ser Ser Leu Ser 85 90
95 Ala Gly Arg Val Arg Arg Met Phe Gly Gly Gly Thr Lys Leu Thr Val
100 105 110 Leu Gly 63 110 PRT Homo sapiens anti-Rh(D) chain O01 63
Ala Glu Leu Thr Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln Arg 1 5
10 15 Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Pro
Tyr 20 25 30 Gly Val His Trp Tyr Gln Gln Phe Pro Gly Thr Ala Pro
Lys Leu Val 35 40 45 Ile Tyr Asn Asp Asn Asn Arg Pro Ser Gly Val
Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys Ser Gly Thr Ser Ala Ser
Leu Ala Ile Thr Gly Leu Gln 65 70 75 80 Ala Glu Asp Glu Ala Asp Tyr
Tyr Cys Gln Ser Tyr Asp Ser Ser Leu 85 90 95 Ser Gly Arg Val Phe
Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110 64 112 PRT Homo
sapiens anti-Rh(D) chain O02 64 Ala Glu Leu Thr Gln Pro Pro Ser Val
Ser Gly Ala Pro Gly Gln Thr 1 5 10 15 Val Thr Ile Ser Cys Thr Gly
Ser Ser Ser Ser Ile Gly Ala Arg Tyr 20 25 30 Asp Val His Trp Tyr
Gln His Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45 Ile Tyr Gly
Asn His Asn Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly
Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu Gln 65 70
75 80 Ala Glu Asp Glu Ala Glu Tyr Tyr Cys Gln Ser Tyr Asp Asn Ser
Leu 85 90 95 Ser Gly Ser Ser Val Phe Phe Gly Gly Gly Thr Lys Leu
Thr Val Leu 100 105 110 65 110 PRT Homo sapiens anti-Rh(D) chain
O03 65 Ala Glu Leu Thr Gln Pro Pro Ser Gly Ala Pro Gly Gln Thr Val
Thr 1 5 10 15 Ile Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly
Tyr Asp Val 20 25 30 His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro
Lys Leu Leu Ile Tyr 35 40 45 Gly Asn Ser Asn Arg Pro Ser Gly Val
Pro Asp Arg Phe Ser Gly Ser 50 55 60 Lys Ser Gly Thr Ser Ala Ser
Leu Ala Ile Thr Gly Leu Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr
Tyr Cys Gln Ser Tyr Asp Ser Ser Leu Ser Gly 85 90 95 Pro Tyr Val
Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110 66 108 PRT
Homo sapiens anti-Rh(D) chain P01 66 Ala Glu Leu Thr Gln Pro Pro
Ser Val Ser Val Ala Pro Arg Gln Thr 1 5 10 15 Ala Arg Ile Thr Cys
Gly Gly Asp Lys Ile Gly Ser Asn Thr Val His 20 25 30 Trp Tyr Arg
Gln Met Ser Gly Gln Ala Pro Val Leu Val Ile Tyr Glu 35 40 45 Asp
Lys Lys Arg Pro Pro Gly Ile Pro Glu Arg Phe Ser Gly Ser Thr 50 55
60 Ser Gly Thr Thr Ala Thr Leu Ser Ile Ser Gly Ala Gln Val Glu Asp
65 70 75 80 Glu Ala Asp Tyr Tyr Cys Tyr Ser Arg Asp Asn Ser Gly Asp
Gln Arg 85 90 95 Arg Val Phe Gly Ala Gly Thr Lys Leu Thr Val Leu
100 105 67 110 PRT Homo sapiens anti-Rh(D) chain Q01 67 Ala Glu Leu
Thr Gln Pro Pro Ser Ala Thr Ala Ser Leu Gly Gly Ser 1 5 10 15 Val
Lys Leu Thr Cys Ile Leu Gln Ser Gly His Arg Asn Tyr Ala Val 20 25
30 Ala Trp His His Gln Glu Ala Gly Lys Gly Pro Arg Phe Leu Met Thr
35 40 45 Val Thr Asn Asp Gly Arg His Ile Lys Gly Asp Gly Ile Pro
Asp Arg 50 55 60 Phe Ser Gly Ser Ala Ser Gly Ala Glu Arg Tyr Leu
Ser Ile Ser Gly 65 70 75 80 Leu Gln Ser Glu Asp Glu Gly Asp Tyr Tyr
Cys Gln Thr Trp Gly Thr 85 90 95 Gly Met His Val Phe Gly Gly Gly
Thr Lys Leu Thr Val Leu 100 105 110 68 108 PRT Homo sapiens
anti-Rh(D) chain R01 68 Ala Glu Leu Thr Gln Pro Pro Ser Ala Ser Gly
Ser Pro Gly Gln Ser 1 5 10 15 Val Thr Ile Ser Cys Thr Gly Ala Ser
Ser Asp Val Gly Ala Tyr Lys 20 25 30 His Val Ser Trp Tyr Gln Gln
His Pro Gly Lys Ala Pro Lys Leu Leu 35 40 45 Thr His Glu Gly Thr
Lys Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60 Gly Ser Lys
Ser Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu Gln 65 70 75 80 Ala
Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Ala Gly Asn Ser 85 90
95 Val Ile Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 69 104
PRT Homo sapiens anti-Rh(D) chain S01 69 Ala Glu Leu Thr Gln Pro
Pro Ser Val Ser Gly Ser Pro Gly Gln Ser 1 5 10 15 Ile Thr Ile Ser
Cys Ser Asp Val Gly Asn Tyr Asn Leu Val Ser Trp 20 25 30 Tyr Gln
Gln Tyr Pro Gly Lys Ala Pro Lys Leu Ile Ile Tyr Glu Gly 35 40 45
Ser Lys Arg Pro Ser Gly Val Ser Ser Arg Phe Ser Gly Ser Arg Ser 50
55 60 Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu Gln Ala Glu Asp
Glu 65 70 75 80 Ala Asp Tyr His Cys Cys Ser Tyr Ala Ile Ser Ser Arg
Ile Phe Gly 85 90 95 Gly Gly Thr Lys Leu Thr Val Leu 100 70 384 DNA
Homo sapiens anti-Rh(D) chain B01 70 gaggtgcagc tgctcgagtc
tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag
cctctggatt caccttcagg agctatgcta tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagct acagcatatg atggaaaaaa taaatactac
180 gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa
cacgctgttt 240 ctgcaaatga acagcctgag agctgaggac acggctgtgt
tttactgtgc gagaggcgga 300 ttttactatg atagtagtgg ttattacggc
ttgaggcact actttgactc ctggggccag 360 ggaaccctgg tcaccgtctc ctca 384
71 372 DNA Homo sapiens anti-Rh(D) chain C03 71 gaggtgcagc
tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt ctccttcagt agctatggca tgcactgggt ccgccaggct
120 ccaggcaagg ggctggagtg ggtgtcagtt atatcatatg atggacatca
taaaaactat 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa aacgctgtac 240 ctgcaaatga acagcctgag acctgaggac
acggctgtat attactgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgc
gtctgttccc tttgatatct ggggcccagg gacaatggtc 360 accgtctctt ca 372
72 372 DNA Homo sapiens anti-Rh(D) chain C01 72 gaggtgcagc
tgctcgagtc ggggggaggt gtggtccagc atgggaggtc cctgagactg 60
tcctgtgcag cctctggatt ctccttcagt agctatggca tgcactgggt ccgccaggct
120 ccaggcaagg ggctggagtg ggtgtcagtt atatcatatg atggacatca
taaaaactat 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa aacgctgtac 240 ctgcaaatga acagcctgag acctgaggac
acggctgtat attactgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgc
gtctgttccc tttgatatat ggggcccagg gacaatggtc 360 accgtgtctt ca 372
73 372 DNA Homo sapiens anti-Rh(D) chain C04 73 gaggtgcagc
tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt ctccttcagt acctatggca tgcactgggt ccgccaggct
120 ccaggcaagg ggctggagtg ggtgtcagtt atatcatatg atggacataa
taaaaactat 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa aacgctgtac 240 ctgcaaatga acagcctgag acctgaggac
acggctgtgt attactgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgc
gtctattcct tttgatatct ggggccaagg gacaatggtc 360 accgtctctt ca 372
74 372 DNA Homo sapiens anti-Rh(D) chain C05 74 gaggtgcagc
tgctcgagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt cagcttcagt agttatggca tgcactgggt ccgccaggct
120 ccaggcaagg ggctggagtg ggtggcagtt atatcgtatg atggaactaa
taaatacttt 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa aacgctgtat 240 ctgcaaatga ccagcctgag acctgaggac
acggctgtgt atttctgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgc
gtccgtacct cttgatatct ggggccaagg gacaatggtc 360 accgtctctt ca 372
75 372 DNA Homo sapiens anti-Rh(D) chain C08 75 gaggtgcagc
tgctcgagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt cagcttcagt agttatggca tgcactgggt ccgccaggct
120 ccaggcaagg ggctggagtg ggtggcagtt atatcgtatg atggaactaa
taaatacttt 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa aacgctgtat 240 ctgcaaatga ccagcctgag acctgaggac
acggctgtgt atttctgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgc
gtctgtacct cttgatatct ggggccaagg gacaatggtc 360 accgtctctt ca 372
76 372 DNA Homo sapiens anti-Rh(D) chain C10 76 gaggtgcagc
tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cctctggatt ctccttcagt agctatggca tgcactgggt ccgccaggct
120 ccaggcaagg ggctggagtg ggtgtcagtt atatcatatg atggacatca
taaaaactat 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa aacgctgtac 240 ctgcaaatga acagcctgag acctgaggac
acggctgtat attactgtgc gaacctaagg 300 ggggaagtaa ctcgtcgtgc
gtctgttccc tttgatatct ggggcccagg gacattggtc 360 accgtctctt ca 372
77 375 DNA Homo sapiens anti-Rh(D) chain D01 77 gaggtgcagc
tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgtag tgtctggttt caccttcaat aactatggca tgcactgggt ccgccaggct
120 ccaggcaagg ggctggagtg ggtggcagtt atttggtttg atggaagtaa
taaatactat 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa cacactgtac 240 ctgcaaatga acagcctgag agccgaggac
acggctgtat attactgtgc gagagagaac 300 cagataaagc tatggtcccg
atacctttac tactttgatt actggggcca gggaaccctg 360 gtcaccgtct cctca
375 78 375 DNA Homo sapiens anti-Rh(D) chain D03 78 gaggtgcagc
tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60
tcctgtgcag cgtctggatt caccttcagt acctatggca tgcactgggt ccgccaggct
120 ccaggcaagg gactggagtg ggtggcagtt atatggtttg atggaagtaa
taaggaatat 180 gcagactccg tgaagggccg attcaccgtc tccagagaca
attccaagaa cacgctgtat 240 ctacaaatga acagcctgag agccgaggac
acggctgtgt attactgtgc gagagaagaa 300 gtggttcggg gagttatctt
atggtctcgg aagtttgact actggggcca gggaaccctg 360 gtcaccgtct cctca
375 79 378 DNA Homo sapiens anti-Rh(D) chain D04 79 gaggtgcagc
tgctcgagtc ggggggaggc gtggcccagc ctgggaggtc cctgagactc 60
tcctgtgtag cgtctggatt cagcctcagg agctatggca tgcactgggt ccgccaggct
120 cctggcaagg ggctggagtg ggtggcagat atatggtttg atggaagtaa
taaagattat 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa cacgttgtat 240 cttcaaatga acagcctgag agccgaggat
acggctgtgt attattgtgc gagagattgg 300 agggtgcggg cctttagtag
tggctggtta agtgcttttg atatctgggg ccaagggaca 360 atggtcaccg tctcctca
378 80 381 DNA Homo sapiens anti-Rh(D) chain D05 80 gaggtgcagc
tgctcgagga gtctggggga ggcgtggccc agcctgggag gtccctgaga 60
ctctcctgtg tagcgtctgg attcagcctc aggagctatg gcatgcactg ggtccgccag
120 gctcctggca aggggctgga gtgggtggca gatatatggt ttgatggaag
taataaagat 180 tatgcagact ccgtgaaggg ccgattcacc atctccagag
acaattccaa gaacacgttg 240 tatcttcaaa tgaacagcct gagagccgag
gacacggctg tgtattattg tgcgagagat 300 tggagggtgc gggcctttag
tagtggctgg ttaagtgctt ttgatatctg gggccaaggg 360 accacggtca
gcgtctcctc a 381 81 375 DNA Homo sapiens anti-Rh(D) chain D07 81
gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc
60 tcctgtgcag tgtctggatt caccctaact aattatggca tgcactgggt
ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcacat gtctggtatg
atggaagtaa aacagaatat 180 gcagactccg tcaagggccg attcgccgtc
tccagagaca aatccaagaa cacactgttt 240 ctgcaaatga acagcctgac
agccgaggac acggctattt attactgtgc gagagagagg 300 agagagaaag
tctatatatt gttctactcg tggctcgacc gctggggcca gggaaccctg 360
gtcaccgtct cctca 375 82 378 DNA Homo sapiens anti-Rh(D) chain D08
82 gaggtgcagc tgctcgagga gtctggggga ggcgtggtcc agcctgggag
gtccctgaga 60 ctctcctgtg cagcgtctgg gttcaccttc agtagctatg
gcatgcactg ggtccgccag 120 gctccaggca gggggctgga gtgggtggct
cttatatggt acgatggagg taacaaagag 180 tatgcagact ccgtgaaggg
ccgcttcagc atctccagag acaattccaa gaacactctg 240 tatctgcaag
tgaacagcct gagagccgac gacacggctg tctattactg tgcgagagac 300
cagagagcag cagcgggtat cttttattat tcccgtatgg acgtctgggg ccaagggacc
360 acggtcaccg tctcctca 378 83 378 DNA Homo sapiens anti-Rh(D)
chain D09 83 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgaag cgtctaaatt caccctctac aattatggca
tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcattt
atatggtttg atggaagtaa taaatactat 180 gaagactccg tgaagggccg
attcaccgtc tccagagaca attccaagaa cacgctgtat 240 ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagagaagga 300
tctaagaagg tggcactttc taggtattac tattatatgg acgtctgggg ccaggggacc
360 acggtcactg tctcgtca 378 84 378 DNA Homo sapiens anti-Rh(D)
chain D10 84 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgaag cgtctaaatt caccctctac aattatggca
tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcattt
atatggtttg atggaagtaa taaatactat 180 gaagactccg tgaagggccg
attcaccgtc tccagagaca attccaagaa cacgctgtat 240 ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagagaagta 300
tctaagaagg tggcactttc taggtattac tactatatgg acgtctgggg ccaggggacc
360 acggtcactg tctcctca 378 85 378 DNA Homo sapiens anti-Rh(D)
chain D11 85 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgaag cgtctaaatt caccctctac aattatggca
tgcactgggt ccgccaggct 120 ccaggcgaag ggctggagtg ggtggcattt
atatggtttg atggaagtaa taaatactat 180 gcagactccg tgaagggccg
attcaccgtc tccagagaca attccaagaa cacgctgtat 240 ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagagaagta 300
tctaagaagc tggcactttc taggtactac tactatatgg acgtctgggg ccaggggacc
360 acggtcactg tctcctca 378 86 378 DNA Homo sapiens anti-Rh(D)
chain D12 86 gaggtgcagc tgctcgagtc ggggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 gcctgtgcag cgtctggatt cagcttcagg agctatggca
tgcactgggt ccgccaggct 120 ccaggcaggg ggctggagtg ggtggcattt
acatggtttg atggaagcaa taaatattat 180 gtagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacgctgtat 240 ctggaaatga
acagcctgag agtcgatgac acggctgtat attactgtgc gagagaggcg 300
tctatgcttc gcggaattag cagatactac tacgcgatgg acgtctgggg cccagggacc
360 acggtcaccg tctcctca 378 87 381 DNA Homo sapiens anti-Rh(D)
chain D13 87 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgcag cgtctggatt caccttcagt acttatggca
tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagtt
atatggtttg atggaagtaa cagagactat 180 gcagagtccg tgaagggccg
attcaccatc tccagagaca agtccaagaa cacactgtat 240 ctgcaaatga
acagcctgag agccgaggac tcggctgtgt attattgtgc gagagaaaat 300
gtggctcgtg gggggggggg cgttcgatac aagtactact ttgactactg gggccaggga
360 accctggtca ccgtctcctc a 381 88 381 DNA Homo sapiens anti-Rh(D)
chain D14 88 gaggtgcagc tgctcgagtc ggggggaggc ttggtacagc ctggggggtc
cctgagactc 60 tcctgtgcag cgtctggatt caccttcagt acttatggca
tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagtt
atatggtttg atggaagtaa gagagactat 180 gcagagtccg tgaagggccg
attcaccatc tccagagaca actccaagaa cacactgtat 240 ctgcaaatga
acagcctgag agccgaggac tcggctgtgt attactgtgc gagagaaaat 300
gtggctcgtg gggggggggg cattcgatac aagtactact ttgactactg gggccaggga
360 accctggtca ccgtctcctc a 381 89 375 DNA Homo sapiens anti-Rh(D)
chain D15 89 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgtag tgtctggatt caccttcaat aactatggca
tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagtt
atttggtttg atggaagtaa taaatactat 180 gcagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacactgtac 240 ctgcaaatga
acagcctgag agccgaggac acggctgtat attactgtgc gagagagaac 300
cagataaagc tatggtcccg atacctttac tactttgact actggggcca gggaaccctg
360 gtcaccgtct cctca 375 90 375 DNA Homo sapiens anti-Rh(D) chain
D16 90 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgtag tgtctggttt caccttcaat aactatggca
tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagtt
atttggtttg atggaagtaa taaatactat 180 gcagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacactgtac 240 ctgcaaatga
acagcctgag agccgaggac acggctgtat attactgtgc gagagagaac 300
cagataaagc tatggtcccg atacctttac tactttgact actggggcca gggaaccctg
360 gtcaccgtct cctca 375 91 375 DNA Homo sapiens anti-Rh(D) chain
D17 91 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgtag tgtctggttt caccttcaat aactatggca
tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagtt
atttggtttg atggaagtaa taaatactat 180 gcagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacactgtac 240 ctgcaaatga
acagcctgag agccgaggac acggctgtat attactgtgc gagagagaac 300
cagataaagc tatggtcccg atacctttac tactttgact actggggcca gggaaccctg
360 gtcaccgtct cctcc 375 92 375 DNA Homo sapiens anti-Rh(D) chain
D18 92 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgtag tgtctggttt caccttcaat aactatggca
tgcactgggt ccgccaggct 120 tcaggcaagg ggttggagtg ggtggcagtt
atttggtttg atggaagtaa taaatactat 180 gcagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacactgtac 240 ctgcaaatga
acagcctgag agccgaggac acggctgtat attactgtgc gagagagaac 300
cagataaagc tatggtcccg atacctttac tactttgact actggggcca gggaaccctg
360 gtcaccgtgt cctca 375 93 375 DNA Homo sapiens anti-Rh(D) chain
D20 93 gaggtgcagc tgctcgagtc ggggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgcag cgtctggatt caccttcagt acctatggca
tgcactgggt ccgccaggct 120 ccaggcaagg gactggagtg ggtggcagtt
atatggtttg atggaagtaa taaggaatat 180 gcagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacgctgtat 240 ctacaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagagaagaa 300
gtggttcggg gagttatctt atggtctcgg aagtttgact actggggcca gggaaccctg
360 gtcaccgtct cctca 375 94 378 DNA Homo sapiens anti-Rh(D) chain
D30 94 gaggtgcagc tgctcgagtc ggggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgcag cgtctggatt caccttcagt agctatggca
tgcgctgggt ccggcaggct 120 ccaggcaagg ggctggagtg ggtggcagtt
gtctactatg atggaagtaa caaacactat 180 tcagactccg tgaagggccg
attcaccatc tccagagaca actccaagaa cacgctgtat 240 ctacaaatgg
acagcctgag agccgaggac acggctgtgt attactgtgc gagagaaaga 300
aattttcgga gtggttattc ccgctactac tacggtatgg acgtctgggg cccagggacc
360 acggtcaccg tctcctca 378 95 378 DNA Homo sapiens anti-Rh(D)
chain D31 95 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60 tcctgtgcag cgtctggatt caccttcagt agctatggca
tgcactgggt ccggcaggct 120 ccaggcaagg ggctggagtg ggtggcagtt
gtctactatg atggaagtaa caaacactat 180 tcagactccg tgaagggccg
attcaccatc tccagagaca actccaagaa cacgctgtat 240 ctacaaatgg
acagcctgag agccgaggac acggctgtgt attactgtgc gagagaaaga 300
aattttcgga gtggttattc ccgctactac tacggtatgg acgtctgggg cccagggacc
360 acggtcaccg tctcctca 378 96 381 DNA Homo sapiens anti-Rh(D)
chain E01 96 gaggtgcagc tgctcgagtc tgggggaggc ctggtcaagc ctggggggtc
cctgagactc 60 tcctgtgcag cctctggatt caccttcagt agctatagca
tgcactgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcatcc
attagtaata gtaatactta catatactac 180 gcagacgcag tgaagggccg
attcaccatc tccagagaca acgccaagaa ctcactgtat 240 ctgcaaatga
acagcctgag agccgaggac acggctgtgt actactgtgc gagagattct 300
agatacagta atttcctccg ttgggttcgg agcgacggta tggacgtctg gggccaaggg
360 accacggtca tcgtctcctc a 381 97 393 DNA Homo sapiens anti-Rh(D)
chain E03 97 gaggtgcagc tgctcgagtc tggggtggag tctgggggag gcctggtcaa
gcctgggggg 60 tccctgagac tctcctgtgc agcctctgga ttcaccttca
gtagctatag catgcactgg 120 gtccgccagg gtccagggaa ggggctggag
tgggtctcat ccattagtaa tagtaatact 180 tacatatact acgcagacgc
agtgaagggc cgattcacca tctccagaga caacgccaag 240 aactcactgt
atctgcaaat gaacagcctg agagccgagc acacggctgt gtactactgt 300
gcgagagatt ctagatacag taatttcctc cgttgggttc ggagcgacgg tatggacgtc
360 tggggccaag ggaccacggt catcgtctcc tca 393 98 321 DNA Homo
sapiens anti-Rh(D) chain F01 98 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcaggg
ctttagaaat gatttaggct ggtatcagca gaaaccaggg 120 aaagccccta
agcgcctgat ctatgctaca tccagtttgc aaagtggggt cccatcaagg 180
ttcagcggca gtggatctgg gacagaattc actctcacaa tcaacagcct gcagcctgaa
240 gattctgcaa cttattactg tctacagcat aatagtttcc cgtggacgtt
cggccaaggg 300 accaaggtgg aaatcaaacg a 321 99 336 DNA Homo sapiens
anti-Rh(D) chain G01 99 gccgagctca ctcagtctcc actctccctg cccgtcaccc
ctggagagcc ggcctccatc 60 tcctgcaggt ctagtcagag cctcctgcat
agtagtggat tcaacttttt ggattggtac 120 ctgcagaagc cagggcagtc
tccacagctc ctgatctata tgggttctaa tcgggcctcc 180 ggggtccctg
acaggttcag tggcagtgga tcaggcacag attttacact gaaaatcaac 240
agagtggagg ctgaggatgt tggggtttat tactgcatgc aagctctaca atttcctctc
300 actttcggcg gagggaccaa ggtggagatc aaacga 336 100 324 DNA Homo
sapiens anti-Rh(D) chain H01 100 gccgagctca cccagtctcc atccttcctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg ccagtcaggg
cattacgagt tatttagcct ggtatcagca aaaaccaggg 120 aaagccccta
agctcctaat ctatgctgca tccactttgc aaagtggggt cccatcaagg 180
ttcagcggca gtggatctgg gacagaattc actctcacaa tcgccagcct gcagcctgat
240 gattttgcaa cttattactg tcaacagctt aataattacc cccctttcac
tttcggccct 300 gggaccaaag tggatatcaa acga 324 101 324 DNA Homo
sapiens anti-Rh(D) chain I01 101 gccgagctca cccagtctcc atcctcccta
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc ctccgtacac
ttttggccag 300 gggaccaagc tggagatcaa acga 324 102 321 DNA Homo
sapiens anti-Rh(D) chain I02 102 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc tgtggacgtt
cggccaaggg 300 accaaggtgg aaatcaaacg a 321 103 321 DNA Homo sapiens
anti-Rh(D) chain I03 103 gccgagctca cccagtctcc atcctccctg
tctgcatctg tagcggacag agtcaccatc 60 acttgccgga caagtcggaa
cattaacaga tacttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ttatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcaccagtct gcaacctgaa
240 gattttgcca cttactactg tcaacagagt tacagtaccc ctttcacttt
cggccctggg 300 accaaagtgg atctcaaacg a 321 104 321 DNA Homo sapiens
anti-Rh(D) chain I04 104 gccgagctca ctcagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagaa
cattaggagg tctttaaatt ggtatcaaca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcagcagagt tccaataccc cgtggacgtt
cggccaaggg 300 accaaggtgg aaatcaaacg a 321 105 321 DNA Homo sapiens
anti-Rh(D) chain I05 105 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattaggagg tatttaaatt ggtatcagca caaaccaggg 120 aaagccccta
agctcctgat ctttgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcactggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc ctcaaacgtt
cggccaaggg 300 accaaggtgg aaatcaaacg a 321 106 321 DNA Homo sapiens
anti-Rh(D) chain I06 106 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgccgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc cgatcacctt
cggccaaggg 300 acacgactgg agattaaacg a 321 107 321 DNA Homo sapiens
anti-Rh(D) chain I07 107 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc ctcgaacttt
cggcggaggg 300 accaaggtgg agatcaaacg a 321 108 321 DNA Homo sapiens
anti-Rh(D) chain I08 108 gccgagctca cccagtctcc attctccctg
tctgcatctg tcggagacag agtcaccata 60 acttgccggg caagtcagac
cattagcagg tctttaaatt ggtatcagca taaaccaggg 120 gaagccccta
agctcctgat ctatgctgca tccagtctgc agcgtggggt cccacccagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gactttgcga cttacttctg tcaacagagt gtcagaatcc cgtacagttt
tggccagggg 300 accaagctgg agatcaaacg a 321 109 321 DNA Homo sapiens
anti-Rh(D) chain I09 109 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagattcc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttattactg tcaacagctt aatagttacc cgtacacttt
tggccagggg 300 accaagctgg agatcaaacg a 321 110 324 DNA Homo sapiens
anti-Rh(D) chain I10 110 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagaa
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc aaagtggggt cctatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc ctccgtatag
ttttggccag 300 gggaccaagc tggagatcaa acga 324 111 309 DNA Homo
sapiens anti-Rh(D) chain I11 111 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
cgctcctgat caatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca ttagcagtct gcaacctgaa
240 gatttcgcaa tttactactg tcaacagaga gaaacttttg gccaggggac
caagctggag 300 atcaaacga 309 112 324 DNA Homo sapiens anti-Rh(D)
chain I12 112 gccgagctca cccagtctcc atcctcccta tctgcatctg
taggagacag agtcaccatc 60 acttgccggg caagtcagag cattagcagc
tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta agctcctgat
ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca
gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240
gattttgcaa cttactactg tcaacagagt tacagtaccc ctccgtacac ttttggccag
300 gggaccaagc tggagatcaa acga 324 113 321 DNA Homo sapiens
anti-Rh(D) chain I13 113 gccgagctca cccagtctcc atcctccctg
tctgcctctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagg tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacggtaccc ctcacagttt
tggccggggg 300 accaagctgg agatcaaacg a 321 114 321 DNA Homo sapiens
anti-Rh(D) chain I15 114 gccgagctca cccagtctcc ttcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caaatcagaa
cattcgtaga tctttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
acctcctgat ctatgctgca tccacattgc aaggtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacttgcg
240 gattttgcaa cttactactg tcaacagact tccgctaccc cgtggacgtt
cggccaaggg 300 accaaggtgg aaatcaaacg a 321 115 321 DNA Homo sapiens
anti-Rh(D) chain I16 115 gccgagctca cccagtctcc atcgtccctg
cctgcatctg tgggagacag agtcaccatc 60 acttgccggg caagtcagac
tattggtttt aatttaaatt ggtatcagca aacatctggg 120 aagcccccta
aactcctaat ctatggtgtt tccaagttgc aaaatggggt cccttcacgg 180
ttcagtggca gtgggtccgg gacggaattc accctcacaa tcagcagtct gcagcctgag
240 gattttgcga cttattattg tcaacagact aacgatgcgt tgtggacgtt
cggccaaggg 300 accaaagtgg aagtcagacg a 321 116 318 DNA Homo sapiens
anti-Rh(D) chain J01 116 gccgagctcc aggaccctgt tgtgtctgtg
gccttgggac agacagtcag gatcacttgc 60 caaggagacg gcctcagaag
ttattatgca agctggtacc agcagaagcc gggacaggcc 120 ccgaaacttg
tcatgtacgg tagaaacaac cggccctcag ggatcccagg ccgattctct 180
ggctccagct cagggcagac agctgccttg accatcacgg ggactcaggc ggaggatgag
240 gctgactatt actgtcagtc ccgtgccacc agcggtaacc ctgtggtgtt
cggcggaggg 300 actaagctga ccgtcctg 318 117 318 DNA Homo sapiens
anti-Rh(D) chain J02 117 gccgagctcc aggaccctgt tgtgtctgtg
gccttgggac agacagtcag gatcacttgc 60 caaggagacg gcctcagaag
ttattatgca agctggtacc agcagaagcc gggacaggcc 120 ccgaaacttg
tcatgtacgg tagaaacaac cggccctcag ggatcccaga ccgattctct 180
ggctccagct cagggcagac agctgccttg accatcacgg ggactcaggc ggaggatgag
240 gctgactatt actgtcagtc ccgtgccacc agcggtaacc ctgtggtgtt
cggcggaggg 300 actaagctga ccgtcctg 318 118 312 DNA Homo sapiens
anti-Rh(D) chain J04 118 gccgagctcc aggaccctgt tgtgtctgtg
gccttgggac agacagtcag gatcacatgc 60 caaggagaca gcctcagaag
ctattatgca agctggtacc agcagaagcc aggacaggcc 120 cctgtacttg
tcatctatgg taaaaacagc cggccctcag ggatcccaga ccgattctct 180
ggctccagct caggaaacac agcttcgttg accatcactg gggctcaggc ggaagatgag
240 gcggactatt attgtagttc gcggggcagc ccccacgtgg cattcggcgg
agggaccaaa 300 ctgaccgtcc tg 312 119 318 DNA Homo sapiens
anti-Rh(D) chain J05 119 gccgagctcc aggaccctgt tgtgtctgtg
gccttgggac agacagtcaa gatcacatgc 60 cagggagaca gcctcagaaa
gtattatgca agctggtacc agcagaagcc aggacaggcc 120 cctgtgcttg
tcttctatgc tagaaatagc cggccctcag ggatcccaga ccgattctct 180
ggctccaact caggaaccac agcttccttg accatcgctg gggctcgggc ggaagatgag
240 gctgactatt actgtcactc ccgggacagc aatggtcacc atcgggtgtt
cggcggaggg 300 accaagctga ccgtccta 318 120 324 DNA Homo sapiens
anti-Rh(D) chain K01 120 gccgagctca ctcaggagcc ctcactgact
gtgtccccag gagggacagt cactctcacc 60 tgtgcttcca gcactggagc
agtcaccagt cgttactttc caaactggtt ccagcagaaa 120 cctggacaag
cacccaggcc actgatttat agtgcaagca acaaacactc ctggacccct 180
gcccggttct caggctccct ccttgggggc aaagctgccc tgacactgtc aggtgtgcag
240 cctgaggacg aggctgagta ttactgcctg ctctactata gtggtgcttg
ggtgttcggc 300 ggagggacca agttgaccgt cctt 324 121 324 DNA Homo
sapiens anti-Rh(D) chain K02 121 gccgagctca ctcaggagcc ctcactgact
gtgtccccag gagggacagt cactctcacc 60 tgtgcttcca gcactggagc
agtcaccagt cgttactttc caaactggtt ccagcagaaa 120 cctggacaag
cacccaggcc actgatttat agtgcaagca acaaacactc ctggacccct 180
gcccggttct caggctccct ccttgggggc aaagctgccc tgacactgtc aggtgtgcag
240 cctgaggacg aggctgagta ttactgcctg ctctactata gtggtgcttg
ggtgttcggc 300 ggagggacca agctgaccgt ccta 324 122 324 DNA Homo
sapiens anti-Rh(D) chain K03 122 gccgagctca ctcagccacc ctcactgact
gtgtccccag gagggacagt cactctcacc 60 tgtgcttcca gcactggagc
agtcaccagt cgttactttc caaactggtt ccagcagaaa 120 cctggccagg
cacccagggc actgatttat ggttcaaaca acaaacactc ctggacccct 180
gcccggttct caggctccct ccttgggggc aaagctgccc tgacactgtc aggtgtgcag
240 cctgaggacg aggcggagta ttactgcctg ctcttctatg ctggtgcttg
ggcgttcggc 300 ggatggacca agctgaccgt ccta 324 123 327 DNA Homo
sapiens anti-Rh(D) chain L01 123 gccgagctca cgcagccgcc ctcagcgtct
gggacccccg ggcagagggt caccatctct 60 tgttctggag gcagctccaa
catcgcaagt aatactgtaa actggtacca gcaactccca 120 ggaacggccc
ccaaactcct catctatagt aataatcagc ggccctcagg ggtccctgac 180
cgattctctg gctccaagtc tggcacctca gccaccctgg tcatcaccgg gctccagact
240 ggggacgagg ccgattatta ctgcggaaca tgggatcaca gccggagtgg
tgcggtgttc 300 ggcggaggga ccaaactgac cgtctta 327 124 327 DNA Homo
sapiens anti-Rh(D) chain L03 124 gccgagctca ctcagccacc ctcagcgtct
gggacccccg ggcagagggt caccatctct 60 tgttctggca gtagctccaa
catcggaaat
aatcatgtaa gctggtacca gcaactccca 120 ggaatggccc ccaaactcct
catctattct aatggtcagc ggccctcagg ggtccctgac 180 cgattctctg
gctccaagtc tggcacctca gcctccctgg ccatcagcgg cctccagtct 240
gaggatgagg ctgattatta ttgtgcagca tggcatgaca gcctctatgg tccggtgttc
300 ggcggaggga ccaagctgac cgtcctc 327 125 327 DNA Homo sapiens
anti-Rh(D) chain L04 125 gccgagctca ctcagccacc ctcagcgtct
gggacccccg ggcagagggt cagcatctct 60 tgttctggaa gcagctccaa
catcggaagt aatactgtaa actggtacca gcagctccca 120 ggaacagccc
ccaaactcct catctctact aataatcagg ggccctcagg agtccctgac 180
cgattctctg gctccaagtc tggcacctca tcctccctgg ccatcagtgg gctccggtca
240 gaggctgagg atgattatta ctgtgcagca tgggatgaca ccctgaatgg
tgtggtattc 300 ggcggaggga ccaaactgac cgtccta 327 126 327 DNA Homo
sapiens anti-Rh(D) chain L05 126 gccgagctca ctcagccacc ctcagcgtct
gggactcccg ggctgagggt caccatctct 60 tgttctggaa gcagctccaa
catcggaagt aatattgtaa actggtacca gcagctccca 120 ggaacggccc
ccaaactcct catctttagt aataataagc ggccctcagg ggtccctgac 180
cgattctctg gctccaagtc tggcacctca gcctccctgg ccatcagtgg gctccagtct
240 gaggatgagg ctgattatta ctgtgctaca tgggatgaca gcctgaatgg
tcgggtgttc 300 ggcggaggga ccaagctgac cgtccta 327 127 327 DNA Homo
sapiens anti-Rh(D) chain M01 127 gccgagctca ctcagccacc ctcagcgtct
gggacccccg ggcagcgggt caccatctct 60 tgttctggga gcaacttcaa
catcggaagt aattatgtat tctggtacca gcatgttcca 120 ggaacggccc
caaaactcct catctataat aataatcaac gcccctctgg ggtccctgac 180
cgactctctg gctccaagtc tggcgcctca gcctccctgg ccatcaatgg gctccggtcc
240 gatgatgagg ctgattatta ctgtacagga tgggatgacc gcctgagtgg
cctgattttc 300 ggcggagggc caaaagtgac cgtccta 327 128 327 DNA Homo
sapiens anti-Rh(D) chain M02 128 gccgagctca cgcagccgcc ctcagcgtct
gggacccccg ggcagagggt caccatctct 60 tgttctggaa gcagctccaa
catcggaagt aattatgtat attggtacca gcagctccca 120 ggaacggccc
ccaaactcct catctatagg aataatcagc ggccctcagg ggtccctgac 180
cgattctctg gctccaagtc tggcacctca gcctccctgg ccatcagtgg gctccggtcc
240 gaggatgagg ctgattatta ctgtgcagca tgggatgaca gcctgagtgg
ttgggtgttc 300 ggcggaggga ccaagctgac cgtccta 327 129 327 DNA Homo
sapiens anti-Rh(D) chain M03 129 gccgagctca ctcagccacc ctcagcgtct
gggacccccg ggcagagggt caccatctct 60 tgttctggaa gcagctccaa
catcggaagt aattatgtat actggtacca gcagctccca 120 ggaacggccc
ccaaactcct catctatagg aataatcagc ggccctcagg ggtccctgac 180
cgattctctg gctccaagtc tggcacctca gcctccctgg ccatcagtgg gctccggtcc
240 gaggctgagg ctgattatta ctgtgcggca tgggatgaca gcctgagtgc
cgtggtattc 300 ggcggaggga ccaaactgac cgtccta 327 130 327 DNA Homo
sapiens anti-Rh(D) chain N01 130 gccgagctca cgcagccgcc ctcagtgtct
gcggccccag gacagaaggt caccatctcc 60 tgctctggaa gcagctccaa
cattgacagt aactatgtat cctggtacca gcagctccca 120 ggaacagccc
ccaaactcct catttttgac aattataggc gaccctcagg gattcctgac 180
cgattctcag gctccaagtc tggcacgtca gccaccctgg gcatcaccgg actccagact
240 ggggacgagg ccgattatta ctgtgcaaca tgggatgaca gcctgaatgg
tcgggtgttc 300 ggcggaggga ccaagctgac cgtccta 327 131 342 DNA Homo
sapiens anti-Rh(D) chain N02 131 gccgagctca cgcagccgcc ctcagtgtct
gcggccccag gacagaaggt caccatctcc 60 tgctctggaa gcagctccaa
cattgggaat aattatgtgt cctggtacca gcaactccca 120 ggaacagccc
ccaaactcct catttatgac aataataagc gaccctcagg gattcctgac 180
cgattctctg gctccaagtc tggcacgtca gccaccctgg gcatcaccgg actccagact
240 ggggacgagg ccgattatta ctgcggaaca tgggatagca gcctgagtgc
tggccgcgtt 300 cggcggatgt tcggcggagg gaccaagttg accgtcctgg gt 342
132 330 DNA Homo sapiens anti-Rh(D) chain O01 132 gccgagctca
cgcagccgcc ctcagtgtct ggggccccag ggcagagggt caccatctcc 60
tgcactggga gcagctccaa catcggggca ccttatggtg tacactggta ccagcagttt
120 ccaggaacag cccccaaact cgtcatctac aatgacaaca atcggccctc
aggggtccct 180 gaccgattct ctggctccaa gtctggcacc tcagcctccc
tggccatcac tgggctccag 240 gctgaggatg aggctgatta ttactgccag
tcctatgaca gcagcctgag tggaagggtg 300 ttcggcggag ggaccaagct
gaccgtccta 330 133 336 DNA Homo sapiens anti-Rh(D) chain O02 133
gccgagctca cgcagccgcc ctcagtgtct ggggccccag ggcagacggt caccatctcc
60 tgcactggga gcagctccag catcggggca cgttatgatg tacactggta
ccaacacctt 120 ccaggaacag cccccaaact cctcatctat ggtaaccaca
atcggccctc aggggtccct 180 gaccgattct ctggctccaa gtctggcacc
tcagcctccc tggccatcac tgggctccag 240 gctgaggatg aggctgaata
ttattgccag tcctatgaca acagcctgag tggttcgtct 300 gtctttttcg
gcggagggac caagctgacc gtccta 336 134 330 DNA Homo sapiens
anti-Rh(D) chain O03 134 gccgagctca cgcagccgcc ctctggggcc
ccaggccaga cggtcaccat ctcctgcact 60 gggagcagct ccaacatcgg
ggcaggttat gatgtacact ggtaccagca gcttccagga 120 acagccccca
aactcctcat ctatggtaac agcaatcggc cctcaggggt ccctgaccga 180
ttctctggct ccaagtctgg cacctcagcc tccctggcca tcactgggct ccaggctgag
240 gatgaggctg attattactg ccagtcctat gacagcagcc tgagtggtcc
ctatgtggta 300 ttcggcggag ggaccaagct gaccgtccta 330 135 324 DNA
Homo sapiens anti-Rh(D) chain P01 135 gccgagctca ctcagccacc
ctcggtgtca gtggccccaa gacagacggc caggattacc 60 tgtggggggg
acaaaatcgg aagtaacact gtgcattggt accggcagat gtcaggccag 120
gcccctgttc tggtcatcta tgaagacaaa aaacgacccc ccgggatccc tgagagattc
180 tctggttcca cctcagggac aacggccacc ttgagtatca gtggggccca
ggttgaggat 240 gaagctgact actactgtta ttcaagagac aacagtggtg
atcagagaag ggtgttcggc 300 gcagggacca agctgaccgt ccta 324 136 330
DNA Homo sapiens anti-Rh(D) chain Q01 136 gccgagctca ctcagccacc
ctccgccact gcctccctgg gaggctcggt caaactcacc 60 tgcattctgc
agagtggcca cagaaattac gccgtcgctt ggcatcacca agaagcaggg 120
aagggcccgc gatttttgat gacggttacc aatgatggca ggcacatcaa gggggacggg
180 atccctgatc gcttctcagg ctccgcctct ggggctgaac gctacctctc
catctccggc 240 ctccagtctg aggatgaggg tgactactac tgtcagacct
ggggcactgg catgcatgtg 300 ttcggcggag ggaccaaact gaccgtccta 330 137
324 DNA Homo sapiens anti-Rh(D) chain R01 137 gccgagctca ctcagcctcc
ctccgcgtcc gggtctcctg gacagtcagt caccatctcc 60 tgcactggag
ccagcagtga cgttggtgct tataagcacg tctcctggta ccaacaacac 120
ccaggcaaag cccccaaact cctgactcat gagggcacta agcggccctc aggggtccct
180 gatcgcttct ctggctccaa gtctggcaac acggcctccc tgaccgtctc
tgggctccag 240 gctgaggatg aggctgatta ttactgcagc tcatttgcag
gtaattccgt gatattcggc 300 ggagggacca agctgaccgt ccta 324 138 312
DNA Homo sapiens anti-Rh(D) chain S01 138 gccgagctca ctcagcctcc
ctccgtgtct gggtctcctg gacagtcgat caccatctcc 60 tgcagtgatg
ttgggaatta taaccttgtc tcctggtacc aacagtaccc aggcaaggcc 120
cccaaactca taatttatga gggcagtaag cggccctcag gggtttctag tcgcttctct
180 ggctccaggt ctggcaacac ggcctccctg acaatctctg ggctccaggc
tgaggacgag 240 gctgattatc actgctgctc atatgcaatt agtagcagga
ttttcggcgg agggaccaag 300 ctgaccgtcc ta 312 139 127 PRT Homo
sapiens anti-Rh(D) antibody clone SH10 139 Glu Val Gln Leu Leu Glu
Glu Ser Gly Gly Gly Val Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg 20 25 30 Asn Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45
Val Ala Phe Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser 50
55 60 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu 65 70 75 80 Tyr Leu Gln Met Asn Ser Leu Arg Ala Asp Asp Thr Ala
Val Tyr Tyr 85 90 95 Cys Ala Arg Glu Glu Ala Leu Phe Arg Gly Leu
Thr Arg Trp Ser Tyr 100 105 110 Gly Met Asp Val Trp Gly Gln Gly Thr
Thr Val Ser Val Ser Ser 115 120 125 140 125 PRT Homo sapiens
anti-Rh(D) antibody clone SH16 140 Glu Val Gln Leu Leu Glu Ser Gly
Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Gly Met His Trp
Val Arg Gln Ala Pro Gly Arg Gly Leu Glu Trp Val 35 40 45 Ala Leu
Ile Trp Tyr Asp Gly Gly Asn Lys Glu Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Ser Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65
70 75 80 Leu Gln Val Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Asp Gln Arg Ala Ala Ala Gly Ile Phe Tyr
Tyr Ser Arg Met 100 105 110 Asp Val Trp Gly Gln Gly Thr Thr Val Thr
Val Ser Ser 115 120 125 141 117 PRT Homo sapiens anti-Rh(D)
antibody clone SH17 141 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu
Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Gly Ala Ser
Gly Ile Pro Phe Val Ser Ser 20 25 30 Trp Met Ala Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Asn Ile Lys Gln
Asp Gly Ser Lys Lys Asn Tyr Val Asp Ser Val 50 55 60 Glu Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr 65 70 75 80 Leu
Gln Met Asp Ser Leu Arg Ala Glu Asp Thr Arg Ile Tyr Tyr Cys 85 90
95 Ala Arg Asp Ser Leu Thr Cys Phe Asp Tyr Trp Gly Gln Gly Ala Leu
100 105 110 Val Thr Val Ser Ser 115 142 128 PRT Homo sapiens
anti-Rh(D) antibody clone SH18 142 Glu Val Gln Leu Leu Glu Ser Gly
Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Arg Ser Tyr 20 25 30 Ala Met His Trp
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala
Thr Ala Tyr Asp Gly Lys Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Met Asn Thr Leu Phe 65
70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Phe
Tyr Cys 85 90 95 Ala Arg Gly Gly Phe Tyr Tyr Asp Ser Ser Gly Tyr
Tyr Gly Leu Arg 100 105 110 His Tyr Phe Asp Ser Trp Gly Gln Gly Thr
Leu Val Thr Val Ser Ser 115 120 125 143 129 PRT Homo sapiens
anti-Rh(D) antibody clone SH20 143 Glu Val Gln Leu Leu Glu Glu Ser
Gly Gly Gly Val Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Arg Ser 20 25 30 Tyr Ala Met His
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45 Val Ala
Val Ile Ser Tyr Asp Gly Ser Thr Ile Tyr Tyr Ala Asp Ser 50 55 60
Val Lys Gly Arg Phe Thr Ile Ser Arg Ala Asn Ser Lys Asn Thr Leu 65
70 75 80 Phe Leu Gln Met Asn Ser Leu Arg Thr Glu Asp Thr Ala Val
Tyr Tyr 85 90 95 Cys Thr Arg Gly Gly Phe Tyr Tyr Asp Ser Ser Gly
Tyr Tyr Gly Leu 100 105 110 Arg His Tyr Phe Asp Tyr Trp Gly Gln Gly
Thr Leu Val Thr Val Ser 115 120 125 Ser 144 126 PRT Homo sapiens
anti-Rh(D) antibody clone SH24 144 Glu Val Gln Leu Leu Glu Ser Gly
Gly Gly Val Ala Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys
Val Ala Ser Gly Phe Ser Leu Arg Ser Tyr 20 25 30 Gly Met His Trp
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Asp
Ile Trp Phe Asp Gly Ser Asn Lys Asp Tyr Ala Asp Ser Val 50 55 60
Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65
70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr Cys 85 90 95 Ala Arg Asp Trp Arg Val Arg Ala Phe Ser Ser Gly
Trp Leu Ser Ala 100 105 110 Phe Asp Ile Trp Gly Gln Gly Thr Met Val
Thr Val Ser Ser 115 120 125 145 127 PRT Homo sapiens anti-Rh(D)
antibody clone SH25 145 Glu Val Gln Leu Leu Glu Glu Ser Gly Gly Gly
Val Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg Leu Ala Cys Ala Ala
Ser Gly Phe Ser Phe Arg Ser 20 25 30 Tyr Gly Met His Trp Val Arg
Gln Ala Pro Gly Arg Gly Leu Glu Trp 35 40 45 Val Ala Phe Thr Trp
Phe Asp Gly Ser Asn Lys Tyr Tyr Val Asp Ser 50 55 60 Val Lys Gly
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu 65 70 75 80 Tyr
Leu Glu Met Asn Ser Leu Arg Val Asp Asp Thr Ala Val Tyr Tyr 85 90
95 Cys Ala Arg Glu Ala Pro Met Leu Arg Gly Ile Ser Arg Tyr Tyr Tyr
100 105 110 Ala Met Asp Val Trp Gly Pro Gly Thr Thr Val Thr Val Ser
Ser 115 120 125 146 126 PRT Homo sapiens anti-Rh(D) antibody clone
SH28, SH50, and SH53 146 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly
Gly Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Asn Ser Tyr 20 25 30 Ala Met Tyr Trp Val Arg
Gln Pro Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Ala Ile Trp
Tyr Asp Gly Ser Asn Lys Glu Tyr Ala Asp Phe Val 50 55 60 Lys Gly
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Ser 65 70 75 80
Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys 85
90 95 Ala Arg Glu Ala Asn Leu Leu Arg Gly Trp Ser Arg Tyr Tyr Tyr
Gly 100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser
Ser 115 120 125 147 126 PRT Homo sapiens anti-Rh(D) antibody clone
SH32 147 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro
Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Glu Ala Ser Lys Phe Thr
Leu Tyr Asn Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly
Lys Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Trp Phe Asp Gly Ser
Asn Lys Tyr Tyr Glu Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg
Glu Leu Ser Lys Lys Val Ala Leu Ser Arg Tyr Tyr Tyr Tyr 100 105 110
Met Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125
148 126 PRT Homo sapiens anti-Rh(D) antibody clone SH37 148 Glu Val
Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15
Ser Leu Arg Leu Ser Cys Glu Ala Ser Lys Phe Thr Leu Tyr Asn Tyr 20
25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45 Ala Phe Ile Trp Phe Asp Gly Ser Asn Lys Tyr Tyr Glu
Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Val Ser Arg Asp Asn Ser
Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Leu Ser Lys Lys
Val Ala Leu Ser Arg Tyr Tyr Tyr Tyr 100 105 110 Met Asp Val Trp Gly
Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 125 149 126 PRT Homo
sapiens anti-Rh(D) antibody clone SH39 149 Glu Val Gln Leu Leu Glu
Gln Ser Gly Gly Gly Val Val Gln Pro Gly 1 5 10 15 Arg Ser Leu Arg
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 20 25 30 Tyr Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 35 40 45
Val Ala Val Ile Trp Phe Asp Gly Ser Asn Lys Glu Tyr Ala Asp Ser 50
55 60 Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu 65 70 75 80 Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr 85 90 95 Cys Ala Arg Glu Glu Val Val Arg Gly Val Ile
Leu Trp Ser Arg Lys 100 105 110 Phe Asp Tyr Trp Gly Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120
125 150 126 PRT Homo sapiens anti-Rh(D) antibody clone SH44 150 Glu
Val Gln Leu Leu Glu Ser Gly Gly Gly Val Ala Gln Pro Gly Arg 1 5 10
15 Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Ser Leu Arg Ser Tyr
20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45 Ala Asp Ile Trp Phe Asp Gly Ser Asn Lys Asp Tyr
Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn
Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Trp Arg Val
Arg Ala Phe Ser Ser Gly Trp Leu Ser Ala 100 105 110 Phe Asp Ile Trp
Gly Gln Gly Thr Met Val Thr Val Ser Ser 115 120 125 151 125 PRT
Homo sapiens anti-Rh(D) antibody clone SH47 151 Glu Val Gln Leu Leu
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg
Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Asn Tyr 20 25 30 Ala
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45 Ala Val Thr Ser Phe Asp Gly Ser Ile Lys Asp Tyr Ala Asp Ser Val
50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Arg Gly Met Ile Val Val Val
Arg Arg Arg Asn Ala Phe 100 105 110 Asp Ile Trp Gly Gln Gly Thr Met
Val Thr Val Ser Ser 115 120 125 152 126 PRT Homo sapiens anti-Rh(D)
antibody clone SH54 152 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Arg 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Thr Phe Ser Arg Asn 20 25 30 Gly Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ala Phe Ile Trp Phe
Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu
Gln Met Asn Ser Leu Arg Ala Asp Asp Thr Ala Val Tyr Tyr Cys 85 90
95 Ala Arg Glu Glu Ala Leu Phe Arg Gly Leu Thr Arg Trp Ser Tyr Gly
100 105 110 Met Asp Val Trp Gly Gln Gly Thr Thr Val Ser Val Ser Ser
115 120 125 153 126 PRT Homo sapiens anti-Rh(D) antibody clone SH56
153 Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg
1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
Ser Tyr 20 25 30 Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45 Ala Val Val Tyr Tyr Asp Gly Ser Asn Lys
His Tyr Ser Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Phe Arg
Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asp Ser Leu
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Arg
Asn Phe Arg Ser Gly Tyr Ser Arg Tyr Tyr Tyr Gly 100 105 110 Met Asp
Val Trp Gly Pro Gly Thr Thr Val Thr Val Ser Ser 115 120 125 154 107
PRT Homo sapiens anti-Rh(D) antibody clone SH8 154 Ala Glu Leu Thr
Gln Ser Pro Ser Ser Leu Ala Ala Ser Val Gly Asp 1 5 10 15 Arg Val
Thr Ile Thr Cys Arg Ala Asn Gln Thr Ile Arg Thr Ser Leu 20 25 30
Asn Trp Tyr Gln Gln Arg Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35
40 45 Gly Ala Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
Gly 50 55 60 Ile Ser Gly Ala Asp Phe Thr Leu Thr Ile Ser Ser Leu
Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Tyr
Gly Tyr Ser Arg Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Asp Ile
Lys Arg 100 105 155 107 PRT Homo sapiens anti-Rh(D) antibody clone
SH12 155 Ala Glu Leu Thr Gln Ser Pro Phe Ser Leu Ser Ala Ser Val
Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser His Asn Ile
Tyr Arg Ser Leu 20 25 30 Asn Trp Phe Gln His Lys Pro Gly Glu Ala
Pro Lys Leu Leu Val Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Arg Gly
Val Pro Thr Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe
Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Ser Ala Thr
Tyr Phe Cys Gln Gln Ser Val Thr Phe Pro Tyr Thr 85 90 95 Phe Gly
Gln Gly Thr Lys Leu Glu Ile Arg Arg 100 105 156 107 PRT Homo
sapiens anti-Rh(D) antibody clone SH13 156 Ala Glu Leu Thr Gln Ser
Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile
Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp
Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45
Ala Ala Ser Ser Leu Arg Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50
55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr
Pro Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg
100 105 157 107 PRT Homo sapiens anti-Rh(D) antibody clone SH14 157
Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5
10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Arg Arg Ser
Leu 20 25 30 Asn Trp Tyr Gln His Lys Pro Gly Arg Ala Pro Arg Leu
Leu Ile Tyr 35 40 45 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser
Arg Phe Arg Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr
Ile Asn Ser Leu Gln Pro Ala 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Ser Ser Asn Thr Pro Trp Thr 85 90 95 Phe Gly His Gly Thr
Lys Val Glu Ile Lys Arg 100 105 158 107 PRT Homo sapiens anti-Rh(D)
antibody clone SH16 158 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser
Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro
Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu
Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp
Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro Thr 85 90
95 Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 100 105 159 106 PRT
Homo sapiens anti-Rh(D) antibody clone SH18 159 Ala Glu Leu Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ile Ala Leu 20 25 30 Asn
Trp Tyr Gln Gln Arg Pro Gly Lys Ala Pro Lys Leu Leu Met Tyr 35 40
45 Ala Thr Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Tyr Asn
Lys Pro Thr Phe 85 90 95 Gly Pro Gly Thr Lys Val Asp Ile Lys Arg
100 105 160 107 PRT Homo sapiens anti-Rh(D) antibody clone SH20 160
Ala Glu Leu Thr Gln Ser Pro Phe Ser Leu Ser Ala Ser Val Gly Asp 1 5
10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Arg Ser
Leu 20 25 30 Asn Trp Tyr Gln His Lys Pro Gly Glu Ala Pro Lys Leu
Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Pro
Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr
Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Phe Cys
Gln Gln Ser Val Arg Ile Pro Tyr Ser 85 90 95 Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys Arg 100 105 161 108 PRT Homo sapiens anti-Rh(D)
antibody clone SH21 161 Ala Glu Leu Thr Gln Ser Pro Ser Phe Leu Ser
Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser
Gln Gly Ile Arg Ser Tyr Leu 20 25 30 Ala Trp Tyr Gln Gln Lys Pro
Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Thr Leu
Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly
Thr Glu Phe Thr Leu Thr Ile Ala Ser Leu Gln Pro Asp 65 70 75 80 Asp
Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Asn Tyr Pro Pro Phe 85 90
95 Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Arg 100 105 162 107
PRT Homo sapiens anti-Rh(D) antibody clone SH24 162 Ala Glu Leu Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Thr Tyr Leu 20 25 30
Asn Trp Tyr Gln Gln Arg Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35
40 45 Ala Ala Ser Thr Leu Gln Arg Gly Val Pro Ser Arg Phe Thr Gly
Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu
Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr
Thr Thr Leu Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Met Glu Ile
Arg Arg 100 105 163 108 PRT Homo sapiens anti-Rh(D) antibody clone
SH26 163 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val
Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile
Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala
Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly
Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe
Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr
Tyr Tyr Cys Gln Gln Ser Tyr Ser Phe Arg Arg Tyr 85 90 95 Ser Phe
Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 100 105 164 107 PRT Homo
sapiens anti-Rh(D) antibody clone SH28 164 Ala Glu Leu Thr Gln Ser
Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile
Thr Cys Arg Ala Asp Gln Asn Ile Arg Arg Ser Leu 20 25 30 Asn Trp
Phe Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45
Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50
55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Ser Thr
Pro Trp Thr 85 90 95 Phe Gly Arg Gly Thr Lys Val Glu Ile Lys Arg
100 105 165 106 PRT Homo sapiens anti-Rh(D) antibody clone SH30 165
Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5
10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Arg Arg Ser
Leu 20 25 30 Asn Trp Tyr Gln Gln Ser Pro Gly Lys Thr Pro Lys Leu
Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser
Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr
Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Ser Tyr Ser Thr Leu Thr Phe 85 90 95 Gly Gly Gly Thr Lys
Val Glu Ile Lys Arg 100 105 166 108 PRT Homo sapiens anti-Rh(D)
antibody clone SH32 166 Ala Glu Leu Thr Gln Glu Pro Ser Leu Thr Val
Ser Pro Gly Gly Thr 1 5 10 15 Val Thr Leu Thr Cys Ala Ser Ser Thr
Gly Ala Val Thr Ser Arg Tyr 20 25 30 Phe Pro Asn Trp Phe Gln Gln
Lys Pro Gly Gln Ala Pro Arg Ala Leu 35 40 45 Ile Tyr Gly Ser Asn
Asn Lys His Ser Trp Thr Pro Ala Arg Phe Ser 50 55 60 Gly Ser Leu
Leu Gly Gly Lys Ala Ala Leu Thr Leu Ser Gly Val Gln 65 70 75 80 Pro
Glu Asp Glu Ala Glu Tyr Tyr Cys Leu Leu Phe Tyr Ala Gly Ala 85 90
95 Trp Ala Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 167 108
PRT Homo sapiens anti-Rh(D) antibody clone SH34 167 Ala Glu Leu Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35
40 45 Ala Ala Ser Gly Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu
Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr
Ser Thr Pro Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Leu Glu
Ile Lys Arg 100 105 168 107 PRT Homo sapiens anti-Rh(D) antibody
clone SH36 168 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser
Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser
Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys
Ser Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser
Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala
Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro Pro Ala 85 90 95 Phe
Gly Pro Gly Thr Lys Val Asp Ile Lys Arg 100 105 169 107 PRT Homo
sapiens anti-Rh(D) antibody clone SH39 169 Ala Glu Leu Thr Gln Ser
Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile
Thr Cys Arg Ala Ser Gln Thr Ile Gly Arg Tyr Leu 20 25 30 Asn Trp
Tyr Gln Gln Arg Pro Gly Lys Ala Pro Lys Leu Leu Val Tyr 35 40 45
Ala Val Ser Ser Leu Gln Ser Gly Ala Pro Ser Arg Phe Ser Gly Ser 50
55 60 Gly Ser Gly Thr His Phe Thr Leu Thr Ile Thr Ser Leu Gln Pro
Glu 65 70 75 80 Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ser Tyr Ser Ser
Pro Phe Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg
100 105 170 107 PRT Homo sapiens anti-Rh(D) antibody clone SH41 170
Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5
10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asn Ile Arg Arg Ser
Leu 20 25 30 Asn Trp Tyr Gln His Lys Pro Gly Arg Ala Pro Arg Leu
Leu Ile Tyr 35 40 45 Ala Ala Ser Thr Leu
Gln Ser Gly Val Pro Ser Arg Phe Arg Gly Ser 50 55 60 Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro Ala 65 70 75 80 Asp
Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Ser Asn Thr Pro Trp Thr 85 90
95 Phe Gly His Gly Thr Lys Val Glu Ile Lys Arg 100 105 171 106 PRT
Homo sapiens anti-Rh(D) antibody clone SH44 171 Ala Glu Leu Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Ile
Ile Thr Cys Arg Ala Ser Gln Thr Ile Pro Arg Phe Leu 20 25 30 Asn
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Val Leu Leu Ile His 35 40
45 Ser Ile Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Ala Ser
50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser
Asn Leu Ser Phe 85 90 95 Gly Pro Gly Thr Thr Val Asp Ile Arg Arg
100 105 172 107 PRT Homo sapiens anti-Rh(D) antibody clone SH46 172
Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5
10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Tyr Ile Ser Ser Tyr
Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu
Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser
Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr
Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Thr Tyr Ser Ser Pro Ser Thr 85 90 95 Phe Gly Pro Gly Thr
Lys Val Asp Ile Lys Arg 100 105 173 107 PRT Homo sapiens anti-Rh(D)
antibody clone SH47 173 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser
Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Ile Thr Cys Arg Ala Ser
Gln Ser Ile Ser Asn Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro
Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu
Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp
Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Tyr Pro Arg Thr 85 90
95 Phe Gly Gln Gly Thr Lys Val Glu Ile Arg Arg 100 105 174 107 PRT
Homo sapiens anti-Rh(D) antibody clone SH48 174 Ala Glu Leu Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Tyr Ile Ser Ser Tyr Leu 20 25 30 Asn
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile Tyr 35 40
45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln
Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Thr Tyr Ser
Ser Pro Ser Thr 85 90 95 Phe Gly Pro Gly Thr Lys Val Asp Ile Lys
Arg 100 105 175 107 PRT Homo sapiens anti-Rh(D) antibody clone SH49
175 Ala Glu Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp
1 5 10 15 Arg Val Thr Val Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser
Tyr Leu 20 25 30 Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys
Leu Leu Ile Tyr 35 40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro
Ser Arg Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr
Cys Gln Gln Ser Tyr Ser Thr Pro Trp Thr 85 90 95 Phe Gly Gln Gly
Thr Lys Val Glu Ile Lys Arg 100 105 176 107 PRT Homo sapiens
anti-Rh(D) antibody clone SH50 176 Ala Glu Leu Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr Val Thr Cys
Arg Ala Ser Gln Ser Ile Ser Ser Tyr Leu 20 25 30 Asn Trp Tyr Gln
Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40 45 Ala Ala
Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser 50 55 60
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro Glu 65
70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Ser Thr Pro
Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100
105 177 108 PRT Homo sapiens anti-Rh(D) antibody clone SH51 177 Ala
Glu Leu Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly Asp 1 5 10
15 Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Ser Tyr Leu
20 25 30 Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile Tyr 35 40 45 Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg
Phe Ser Gly Ser 50 55 60 Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile
Ser Ser Leu Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln
Gln Leu Asn Asn Tyr Pro Pro Phe 85 90 95 Thr Phe Gly Pro Gly Thr
Lys Val Asp Ile Lys Arg 100 105 178 108 PRT Homo sapiens anti-Rh(D)
antibody clone SH52 178 Ala Glu Leu Thr Gln Ser Pro Gly Thr Leu Ser
Leu Ser Pro Gly Glu 1 5 10 15 Arg Ala Thr Leu Ser Cys Arg Ala Ser
Gln Ser Ile Ser Ser Ser Tyr 20 25 30 Leu Ala Trp Tyr Gln Gln Lys
Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45 Tyr Gly Ala Ser Ser
Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser Gly 50 55 60 Ser Gly Ser
Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu Pro 65 70 75 80 Glu
Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro Trp 85 90
95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg 100 105 179 107
PRT Homo sapiens anti-Rh(D) antibody clone SH54 179 Ala Glu Leu Thr
Gln Ser Pro Ser Ser Met Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Gly Thr Tyr Leu 20 25 30
Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35
40 45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
Ser 50 55 60 Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu
Gln Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr
Ser Thr Pro Trp Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile
Lys Arg 100 105 180 109 PRT Homo sapiens anti-Rh(D) antibody clone
SH55 180 Ala Glu Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly
Gln Arg 1 5 10 15 Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile
Gly Ser Lys Tyr 20 25 30 Val Tyr Trp Tyr Gln Gln Leu Pro Gly Thr
Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Ser Asn Asn Gln Arg Pro Ser
Gly Val Pro Asp Arg Phe Ser Ala 50 55 60 Phe Lys Ser Gly Thr Ser
Ala Ser Leu Ala Ile Thr Gly Leu Gln Ala 65 70 75 80 Glu Asp Glu Ala
Asn Tyr Tyr Cys Gln Ser Tyr Asp Ser Gly Leu Ser 85 90 95 Gly Trp
Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 181 108 PRT
Homo sapiens anti-Rh(D) antibody clone SH56 181 Ala Glu Leu Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly Asp 1 5 10 15 Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Arg Tyr Leu 20 25 30 Asn
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile Tyr 35 40
45 Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly Ser
50 55 60 Gly Ser Gly Thr Asp Phe Ala Leu Thr Ile Ser Ser Leu Leu
Pro Glu 65 70 75 80 Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gly Tyr Ser
Thr Pro Pro Tyr 85 90 95 Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg 100 105 182 381 DNA Homo sapiens anti-Rh(D) antibody clone
SH10 182 gaggtgcagc tgctcgagga gtctggggga ggcgtggtcc agcctgggag
gtccctgaga 60 ctctcctgtg cagcgtctgg gttcaccttc agtaggaatg
gcatgcactg ggtccgccag 120 gctcctggca aggggctgga gtgggtggca
tttatatggt ttgatggaag taataaatac 180 tatgcagact ccgtgaaggg
ccgattcacc atctccagag acaattccaa gaacacgctg 240 tatctgcaaa
tgaacagcct gagagccgac gacacggctg tgtattactg tgcgagagag 300
gaggctctgt ttcggggact tactcggtgg tcctacggca tggacgtctg gggccaaggg
360 accacggtca gcgtctcctc a 381 183 375 DNA Homo sapiens anti-Rh(D)
antibody clone SH16 183 gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc
ctgggaggtc cctgagactc 60 tcctgtgcag cgtctgggtt caccttcagt
agctatggca tgcactgggt ccgccaggct 120 ccaggcaggg ggctggagtg
ggtggctctt atatggtacg atggaggtaa caaagagtat 180 gcagactccg
tgaagggccg cttcagcatc tccagagaca actccaagaa cactctgtat 240
ctgcaagtga acagcctgag agccgacgac acggctgtct attactgtgc gagagaccag
300 agagcagcag cgggtatctt ttattattcc cgtatggacg tctggggcca
agggaccacg 360 gtcaccgtct cctca 375 184 351 DNA Homo sapiens
anti-Rh(D) antibody clone SH17 anti-Rh(D) antibody clone SH17 184
gaggtgcagc tgctcgagtc tgggggaggc ttggtccagc cgggggggtc cctgagactc
60 tcctgtggtg cctctggaat cccctttgtt tcctcttgga tggcctgggt
ccgccaggcc 120 ccagggaagg ggctggagtg ggtggccaac ataaaacaag
atggaagtaa gaaaaactat 180 gtggactctg tggagggccg attcaccatc
tccagagaca acgcgaagaa ctcactttat 240 ctgcaaatgg acagcctgag
agccgaggac acgcggatat attactgtgc gcgagattca 300 cttacttgtt
ttgactactg gggccaggga gccctggtca ccgtctcctc a 351 185 384 DNA Homo
sapiens anti-Rh(D) antibody clone SH18 185 gaggtgcagc tgctcgagtc
tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag
cctctggatt caccttcagg agctatgcta tgcactgggt ccgccaggct 120
ccaggcaagg ggctggagtg ggtggcagct acagcatatg atggaaaaaa taaatactac
180 gcagactccg tgaagggccg attcaccatc tccagagaca attccatgaa
cacgctgttt 240 ctgcaaatga acagcctgag agctgaggac acggctgtgt
tttactgtgc gagaggcgga 300 ttttactatg atagtagtgg ttattacggc
ttgaggcact actttgactc ctggggccag 360 ggaaccctgg tcaccgtctc ctca 384
186 387 DNA Homo sapiens anti-Rh(D) antibody clone SH20 186
gaggtgcagc tgctcgagga gtctggggga ggcgtggtcc agcctgggag gtccctgaga
60 ctctcctgtg cagcctctgg attcaccttc agaagttatg ctatgcactg
ggtccgccag 120 gctccaggca aggggctgga gtgggtggcg gttatatcat
atgatggaag tactatatac 180 tacgcagact ccgtgaaggg ccgattcacc
atctccagag ccaattccaa gaacacgctg 240 tttctgcaaa tgaacagcct
cagaactgag gacacggctg tatattactg tacgagaggg 300 gggttttact
atgacagtag tggttattac gggttgaggc actactttga ctactggggc 360
cagggaaccc tggtcaccgt ctcttca 387 187 378 DNA Homo sapiens
anti-Rh(D) antibody clone SH24 187 gaggtgcagc tgctcgagtc ggggggaggc
gtggcccagc ctgggaggtc cctgagactc 60 tcctgtgtag cgtctggatt
cagcctcagg agctatggca tgcactgggt ccgccaggct 120 cctggcaagg
ggctggagtg ggtggcagat atatggtttg atggaagtaa taaagattat 180
gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgttgtat
240 cttcaaatga acagcctgag agccgaggac acggctgtgt attattgtgc
gagagattgg 300 agggtgcggg cctttagtag tggctggtta agtgcttttg
atatctgggg ccaagggaca 360 atggtcaccg tctcttca 378 188 381 DNA Homo
sapiens anti-Rh(D) antibody clone SH25 188 gaggtgcagc tgctcgagga
gtctggggga ggcgtggtcc agcctgggag gtccctgaga 60 ctcgcctgtg
cagcgtctgg attcagcttc aggagctatg gcatgcactg ggtccgccag 120
gctccaggca gggggctgga gtgggtggca tttacatggt ttgatggaag caataaatat
180 tatgtagact ccgtgaaggg ccgattcacc atctccagag acaattccaa
gaacacgctg 240 tatctggaaa tgaacagcct gagagtcgat gacacggctg
tatattactg tgcgagagag 300 gcgcctatgc ttcgcggaat tagcagatac
tactacgcga tggacgtctg gggcccaggg 360 accacggtca ccgtctcctc a 381
189 378 DNA Homo sapiens anti-Rh(D) antibody clone SH28, SH50, and
SH53 189 gaggtgcagc tgctcgagtc tgggggaggc ggggtccagc ctgggaggtc
cctgcgactc 60 tcctgtgcgg cgtctggatt caccttcaat agttatgcca
tgtactgggt ccgccagcct 120 ccaggcaagg ggctggagtg ggtggcagct
atatggtatg atggaagtaa taaagaatat 180 gcagattttg tgaagggccg
cttcaccatc tccagagaca attccaagaa cacgctgtct 240 ctgcaaatga
acagcctgag agacgaggac acggctgtgt attactgtgc gagagaggcg 300
aatctcctcc gtggctggtc tcgatactac tacggtatgg acgtctgggg ccaagggacc
360 acggtcaccg tctcctca 378 190 378 DNA Homo sapiens anti-Rh(D)
antibody clone SH32 190 gaggtgcagc tgctcgagtc ggggggaggc gtggtccagc
ctgggaggtc cctgagactc 60 tcctgtgaag cgtctaaatt caccctctac
aattatggca tgcactgggt ccgccaggct 120 ccaggcaagg ggctggagtg
ggtggcattt atatggtttg atggaagtaa taaatactat 180 gaagactccg
tgaagggccg attcaccgtc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagagaacta
300 tctaagaagg tggcactttc taggtattac tactatatgg acgtctgggg
ccaggggacc 360 acggtcactg tctcgtca 378 191 378 DNA Homo sapiens
anti-Rh(D) antibody clone SH37 191 gaggtgcagc tgctcgagga gtctggggga
ggcgtggtcc agcctgggag gtccctgaga 60 ctctcctgtg cagtgtctgg
attcacccta actaattatg gcatgcactg ggtccgccag 120 gctccaggca
aggggctgga gtgggtggca catgtctggt atgatggaag taaaacagaa 180
tacgcagact ccgtcaaggg ccgattcgcc gtctccagag acaaatccaa gaacacactg
240 tttctgcaaa tgaacagcct gacagccgag gacacggcta tttattactg
tgcgagagag 300 aggagagaga aagtctatat attgttctac tcgtggctcg
accgctgggg ccagggaacc 360 ctggtcaccg tctcctca 378 192 378 DNA Homo
sapiens anti-Rh(D) antibody clone SH39 192 gaggtgcagc tgctcgagca
gtctggggga ggcgtggtcc agcctgggag gtccctgaga 60 ctctcctgtg
cagcgtctgg attcaccttc agtagctatg gcatgcactg ggtccgccag 120
gctccaggca agggactgga gtgggtggca gttatatggt ttgatggaag taataaggaa
180 tatgcagact ccgtgaaggg ccgattcacc atctccagag acaattccaa
gaacacgctg 240 tatctacaaa tgaacagcct gagagccgag gacacggctg
tgtattactg tgcgagagaa 300 gaagtggttc ggggagttat cttatggtct
cggaagtttg actactgggg ccagggaacc 360 ctggtcaccg tctcctca 378 193
378 DNA Homo sapiens anti-Rh(D) antibody clone SH44 193 gaggtgcagc
tgctcgagtc ggggggaggc gtggcccagc ctgggaggtc cctgagactc 60
tcctgtgtag cgtctggatt cagcctcagg agctatggca tgcactgggt ccgccaggct
120 cctggcaagg ggctggagtg ggtggcagat atatggtttg atggaagtaa
taaagattat 180 gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa cacgttgtat 240 cttcaaatga acagcctgag agccgaggat
acggctgtgt attattgtgc gagagattgg 300 agggtgcggg cctttagtag
tggctggtta agtgcttttg atatctgggg ccaagggaca 360 atggtcaccg tctcttca
378 194 375 DNA Homo sapiens anti-Rh(D) antibody clone SH47 194
gaggtgcagc tgctcgagtc tgggggaggc gtggtccagc ctgggaggtc cctgcgactc
60 tcttgtgcag cctctggatt cagcttcagt aactatgcta tgcactgggt
ccgccaggct 120 ccaggcaagg ggctggagtg ggtggcagtt acatcatttg
atggaagcat taaagactac 180 gcagactccg tgaagggccg attcaccatc
tccagagaca attccaagaa cacactatat 240 ctgcaaatga acagcctgag
agatgaggac acggctgtat attactgtgc gagagagcgg 300 gggatgatag
tcgtggtccg tcgcagaaat gcttttgata tttggggcca agggacaatg 360
gtcaccgtct cttca 375 195 378 DNA Homo sapiens anti-Rh(D) antibody
clone SH54 195 gaggtgcagc tgctcgagtc ggggggaggc gtggtccagc
ctgggaggtc cctgagactc 60 tcctgtgcag cgtctgggtt caccttcagt
aggaatggca tgcactgggt ccgccaggct 120 cctggcaagg ggctggagtg
ggtggcattt atatggtttg atggaagtaa taaatactat 180 gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240
ctgcaaatga acagcctgag agccgacgac acggctgtgt attactgtgc gagagaggag
300 gctctgtttc ggggacttac tcggtggtcc tacggtatgg acgtctgggg
ccaagggacc 360 acggtcagcg tctcctca 378 196 378 DNA Homo sapiens
anti-Rh(D) antibody clone SH56 196 gaggtgcagc tgctcgagtc ggggggaggc
gtggtccagc ctgggaggtc cctgagactc 60 tcctgtgcag cgtctggatt
caccttcagt agctatggca tgcactgggt ccggcaggct 120 ccaggcaagg
ggctggagtg
ggtggcagtt gtctactatg atggaagtaa caaacactat 180 tcagactccg
tgaagggccg attcaccatc ttcagagaca actccaagaa cacgctgtat 240
ctacaaatgg acagcctgag agccgaggac acggctgtgt attactgtgc gagagaaaga
300 aattttcgga gtggttattc ccgctactac tacggtatgg acgtctgggg
cccagggacc 360 acggtcaccg tctcctca 378 197 321 DNA Homo sapiens
anti-Rh(D) antibody clone SH8 197 gccgagctca cccagtctcc atcctccctg
gctgcgtctg tcggagacag agtcaccatc 60 acttgccggg caaatcagac
catcagaacc tctttaaatt ggtatcaaca aagacctggg 120 aaagccccta
acctcctgat ctatggtgca tccaggttgc atagtggggt cccatcaagg 180
tttagtggcg gtatttctgg ggcagacttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcagcagact tacggttatt ctcgaacgtt
cggccaaggg 300 accaaggtgg atatcaaacg a 321 198 321 DNA Homo sapiens
anti-Rh(D) antibody clone SH12 198 gccgagctca cccagtctcc attctccctg
tctgcatctg taggagacag agtcaccata 60 acttgccggg caagtcacaa
catttacagg tctttaaatt ggtttcagca taaaccaggg 120 gaagccccta
agctcctggt ctatgctgca tccagtctgc agcgtggggt cccaaccagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct tcaacctgaa
240 gactctgcga cttacttctg tcaacagagt gtcacattcc cctacacttt
tggccagggg 300 accaagctgg agatcagacg a 321 199 321 DNA Homo sapiens
anti-Rh(D) antibody clone SH13 199 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc gaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc cctacacttt
tggccagggg 300 accaagctgg agatcaaacg a 321 200 321 DNA Homo sapiens
anti-Rh(D) antibody clone SH14 200 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagaa
cattaggagg tctttaaatt ggtatcaaca caaaccaggg 120 agagccccta
gactcctgat ctatgctgca tccactttgc aaagtggggt cccatcaagg 180
ttcaggggca gtggatctgg gacagatttc actctcacca tcaacagtct gcaacctgca
240 gattttgcaa cttactactg tcagcagagt tccaataccc cgtggacgtt
cggccatggg 300 accaaggtgg aaatcaaacg a 321 201 321 DNA Homo sapiens
anti-Rh(D) antibody clone SH16 201 gccgagctca cccagtctcc atcctccctg
tctgcctctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcaaca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc ctccaacttt
cggcggaggg 300 accaaggtgg agatcaaacg a 321 202 318 DNA Homo sapiens
anti-Rh(D) antibody clone SH18 202 gccgagctca cccagtctcc atcctccctc
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
tattagcatc gctttaaatt ggtatcagca gagaccaggg 120 aaagccccta
agctcctgat gtatgctaca tccactttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacaatat tacaataaac ctactttcgg
ccctgggacc 300 aaggtggata tcaaacga 318 203 321 DNA Homo sapiens
anti-Rh(D) antibody clone SH20 203 gccgagctca cccagtctcc attctccctg
tctgcatctg tcggagacag agtcaccata 60 acttgccggg caagtcagag
cattagcagg tctttaaatt ggtatcaaca taaaccaggg 120 gaagccccta
agctcctgat ctatgctgca tccagtctgc agcgtggggt cccacccagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gactttgcga cttacttctg tcaacagagt gtcagaatcc cgtacagttt
tggccagggg 300 accaagctgg agatcaaacg a 321 204 324 DNA Homo sapiens
anti-Rh(D) antibody clone SH21 204 gccgagctca cccagtctcc atccttcctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg ccagtcaggg
cattaggagt tatttagcct ggtatcagca aaaaccaggg 120 aaagccccta
agctcctaat ctatgctgca tccactttgc aaagtggggt cccatcaagg 180
ttcagcggca gtggatctgg gacagaattc actctcacaa tcgccagcct gcagcctgat
240 gattttgcaa cttattactg tcaacagctt aataattacc cccctttcac
tttcggccct 300 gggaccaaag tggatatcaa acga 324 205 321 DNA Homo
sapiens anti-Rh(D) antibody clone SH24 205 gccgagctca cccagtctcc
atcctccctg tctgcatctg taggagacag agtcaccatc 60 acttgccggg
caagtcagag cattagcacc tatttaaatt ggtatcagca gagaccaggg 120
aaagccccta acctcctgat ctatgctgca tccactttgc aaaggggggt cccatcaagg
180 ttcactggca gtggatctgg gacagatttc actctcacca tcagcagtct
gcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tacactaccc
tgtggacgtt cggccaaggg 300 accaagatgg aaatcagacg a 321 206 324 DNA
Homo sapiens anti-Rh(D) antibody clone SH26 206 gccgagctca
cccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc 60
acttgccggg caagtcagag cattagcagc tatttaaatt ggtatcagca gaaaccaggg
120 aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggt
cccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca
tcagcagtct gcaacctgaa 240 gattttgcaa cttactactg tcaacagagt
tacagtttcc gaaggtacag ttttggccag 300 gggaccaagc tggagatcaa acga 324
207 321 DNA Homo sapiens anti-Rh(D) antibody clone SH28 207
gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc
60 acttgccggg cagatcagaa cattaggagg tctttaaatt ggtttcagca
gaaaccaggg 120 aaagccccta agctcctgat ctatgctgca tccagtttgc
aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg gacagatttc
actctcacca tcagcagtct gcaacctgaa 240 gattttgcaa cttactactg
tcaacagagt tccagtaccc cgtggacgtt cggccgaggg 300 accaaggtgg
aaatcaaacg a 321 208 318 DNA Homo sapiens anti-Rh(D) antibody clone
SH30 208 gccgagctca cccagtctcc atcctccctg tctgcatctg ttggagacag
agtcaccatc 60 acttgccggg caagtcagag cattcggagg tctttaaatt
ggtatcagca gagtccaggg 120 aaaaccccta agctcctgat ctatgctgca
tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca gtggatctgg
gacagatttc actctcacca tcagcagtct gcaacctgaa 240 gattttgcaa
cttactactg tcaacagagt tacagtaccc tcactttcgg cggagggacc 300
aaggtggaga tcaaacga 318 209 324 DNA Homo sapiens anti-Rh(D)
antibody clone SH32 209 gccgagctca ctcaggagcc ctcactgact gtgtccccag
gagggacagt cactctcacc 60 tgtgcttcca gcactggagc agtcaccagt
cgttactttc caaactggtt ccagcagaaa 120 cctggccagg cacccagggc
actgatttat ggttcaaaca acaaacactc ctggacccct 180 gcccggttct
caggctccct ccttgggggc aaagctgccc tgacactgtc aggtgtgcag 240
cctgaggacg aggcggagta ttactgcctg ctcttctatg ctggtgcttg ggcgttcggc
300 ggagggacca agctgaccgt ccta 324 210 324 DNA Homo sapiens
anti-Rh(D) antibody clone SH34 210 gccgagctca cccagtctcc atcctccctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg caagtcagag
cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta
agctcctgat ctatgctgca tccggtttgc aaagtggggt cccatcaagg 180
ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa
240 gattttgcaa cttactactg tcaacagagt tacagtaccc ccccgtacac
ttttggccag 300 gggaccaagc tggagatcaa acga 324 211 321 DNA Homo
sapiens anti-Rh(D) antibody clone SH36 211 gccgagctca ctcagtctcc
atcctccctg tctgcatctg taggagacag agtcaccatc 60 acttgccggg
caagtcagag cattagcagc tatttaaatt ggtatcagca gaaaccaggg 120
aaatccccta agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg
180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct
gcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tacagtaccc
ctccggcttt cggccctggg 300 accaaagtgg atatcaaacg a 321 212 321 DNA
Homo sapiens anti-Rh(D) antibody clone SH39 212 gccgagctca
cccagtctcc atcctccctg tctgcatctg tgggagacag agtcaccatc 60
acttgccggg caagtcagac cattgggagg tatttaaatt ggtatcagca gaggccaggg
120 aaagccccca aactcctggt atatgctgtg tccagtttgc aaagtggggc
cccatcaagg 180 ttcagtggca gtggctctgg gacacatttc actctcacca
tcaccagtct gcaacctgaa 240 gattttgcaa cttacttctg ccaacagagt
tacagttctc ctttcacttt tggccagggg 300 accaaggttg agatcaaacg a 321
213 321 DNA Homo sapiens anti-Rh(D) antibody clone SH41 213
gccgagctca cccagtctcc atcctccctg tctgcatctg taggagacag agtcaccatc
60 acttgccggg caagtcagaa cattaggagg tctttaaatt ggtatcaaca
caaaccaggg 120 agagccccta gactcctgat ctatgctgca tccactttgc
aaagtggggt cccatcaagg 180 ttcaggggca gtggatctgg gacagatttc
actctcacca tcaacagtct gcaacctgca 240 gattttgcaa cttactactg
tcagcagagt tccaataccc cgtggacgtt cggccatggg 300 accaaggtgg
aaatcaaacg a 321 214 318 DNA Homo sapiens anti-Rh(D) antibody clone
SH44 214 gccgagctca cccagtctcc atcgtccctg tctgcatctg taggagacag
agtcatcatc 60 acttgccggg caagtcagac cattcccagg ttcttgaatt
ggtatcaaca gaagcctgga 120 aaagcccctg ttctcctgat tcatagtata
tccagtttac aaagtggggt cccatcaagg 180 ttcagtgcca gtggatctgg
gacagagttc actctcacca tcagcagtct gcaacctgaa 240 gatttcgcaa
cttactactg ccaacagagt tacagtaatc tctctttcgg ccctgggacc 300
acagtggata ttagacga 318 215 321 DNA Homo sapiens anti-Rh(D)
antibody clone SH46 215 gccgagctca cccagtctcc atcctccctg tctgcatctg
taggagacag agtcaccatc 60 acttgccggg caagtcagta cattagcagc
tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta atctcctgat
ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca
gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240
gattttgcaa cttactactg tcaacagact tacagttccc ctagcacttt cggccctggg
300 accaaagtgg atatcaaacg a 321 216 321 DNA Homo sapiens anti-Rh(D)
antibody clone SH47 216 gccgagctca cccagtctcc atcctccctg tctgcatctg
taggagacag agtcaccatc 60 acttgccggg caagtcagag cattagcaac
tatttaaatt ggtatcagca gaaaccagga 120 aaagccccta acctcctgat
ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca
gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240
gattttgcaa cttactactg tcaacagagt tacagttatc ctcgcacgtt cggccaaggg
300 accaaggtgg agatcagacg a 321 217 321 DNA Homo sapiens anti-Rh(D)
antibody clone SH48 217 gccgagctca cccagtctcc atcctccctg tctgcatctg
taggagacag agtcaccatc 60 acttgccggg caagtcagta cattagcagc
tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta atctcctgat
ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca
gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240
gattttgcaa cttactactg tcaacagact tacagttccc ctagcacttt cggccctggg
300 accaaagtgg atatcaaacg a 321 218 321 DNA Homo sapiens anti-Rh(D)
antibody clone SH49 218 gccgagctca cccagtctcc atcctccctg tctgcatctg
taggagacag agtcaccgtc 60 acttgccggg caagtcagag cattagcagc
tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccta agctcctgat
ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca
gtggatctgg gacagatttc actctcacca tcagcagtct gcaacctgaa 240
gattttgcaa cttactactg tcaacagagt tacagtaccc cgtggacgtt cggccaaggg
300 accaaggtgg aaatcaaacg a 321 219 324 DNA Homo sapiens anti-Rh(D)
antibody clone SH50 219 gccgagctca cccagtctcc atcgtccctg tctgcatctg
taggagacag agtcaccatc 60 acttgccgga caagtcagag cattggcacc
tatttaaatt ggtatcaaca aaaaccaggg 120 aaagccccta aactcctgat
ctatgctgca tccaatgtgc aaagtggggt cccatcaagg 180 ttcagtggcg
gtggatctgg gacaggtttc tctctcatca tcagcagtct gcaacctgaa 240
gatttagcaa tttactactg ccaacagagc tacagtgtcc ctccgtacag ctttggcccg
300 gggaccaagc tggagatcaa acga 324 220 324 DNA Homo sapiens
anti-Rh(D) antibody clone SH51 220 gccgagctca cacagtctcc atccttcctg
tctgcatctg taggagacag agtcaccatc 60 acttgccggg ccagtcaggg
cataaggagt tatttagcct ggtatcagca aaaaccaggg 120 aaagccccta
agctcctaat ctatgctgca tccactttgc aaagtggggt cccatcaagg 180
ttcagcggca gtggatctgg gacagaattc actctcacaa tcagcagcct gcagcctgaa
240 gattttgcaa cttattactg tcaacagctt aataattacc cccctttcac
tttcggccct 300 gggaccaaag tggatatcaa acga 324 221 321 DNA Homo
sapiens anti-Rh(D) antibody clone SH52 221 gccgagctca cccagtctcc
atcctccatg tctgcatctg taggagacag agtcaccatc 60 acttgccggg
caagtcagag cattggcact tatttaaatt ggtatcagca gaaaccaggg 120
aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggt cccatcaagg
180 ttcagtggca gtggatctgg gacagatttc actctcacca tcagcagtct
gcaacctgaa 240 gattttgcaa cttactactg tcaacagagt tacagtaccc
cgtggacgtt cggccaaggg 300 accaaggtgg aaatcaaacg a 321 222 321 DNA
Homo sapiens anti-Rh(D) antibody clone SH54 222 gccgagctca
cccagtctcc atcctccatg tctgcatctg taggagacag agtcaccatc 60
acttgccggg caagtcagag cattggcact tatttaaatt ggtatcagca gaaaccaggg
120 aaagccccta agctcctgat ctatgctgca tccagtttgc aaagtggggt
cccatcaagg 180 ttcagtggca gtggatctgg gacagatttc actctcacca
tcagcagtct gcaacctgaa 240 gattttgcaa cttactactg tcaacagagt
tacagtaccc cgtggacgtt cggccaaggg 300 accaaggtgg aaatcaaacg a 321
223 327 DNA Homo sapiens anti-Rh(D) antibody clone SH55 223
gccgagctca cgcagccgcc ctcagcgtct gggacccccg ggcagagggt caccatctct
60 tgttctggaa gcagctccaa catcggaagt aaatatgtat actggtacca
gcaactccca 120 ggaacggccc ccaaactcct catttatagt aataatcagc
ggccctcagg ggtccctgac 180 cgattctctg ccttcaagtc tggcacctca
gcctccctgg ccatcactgg gctccaggct 240 gaggatgagg ctaattatta
ctgccagtcc tatgacagcg gcctgagtgg ctgggtgttc 300 ggcggcggga
ccaagctgac cgtccta 327 224 324 DNA Homo sapiens anti-Rh(D) antibody
clone SH56 224 gccgagctca cccagtctcc atcctccctg tctgcatctg
taggagacag agtcaccatc 60 acttgccggg caagtcagag cattagcagg
tatttaaatt ggtatcagca gaaaccaggg 120 aaagccccca agctcctgat
ctatgctgca tccagtttgc aaagtggggt cccatcaagg 180 ttcagtggca
gtggatctgg gacagatttc gctctcacca tcagcagtct gctacctgaa 240
gattttgcaa cttactactg tcaacagggt tacagtaccc ctccgtacag ttttggccag
300 gggaccaagc tggagatcaa acga 324
* * * * *
References