Erythropoietin Receptor Binding Antibodies

Abstract

The present invention relates to antibodies and antibody fragments thereof that bind to and activate an erythropoietin receptor. The present invention also relates to methods of modulating the endogenous activity of an erythropoietin receptor in a mammal using said antibodies as well as pharmaceutical compositions containing said antibodies.

Description

APPLICATION HISTORY

[0001] This application claims priority to U.S. Provisional Application Ser. No. 60/418,031, filed Oct. 14, 2002, hereby incorporated by reference in its entirety.

TECHNICAL FIELD OF THE INVENTION

[0002] The present invention relates to antibodies that recognize, bind to and, preferably, activate the erythropoietin receptor.

BACKGROUND OF THE INVENTION

[0003] Erythropoietin ("EPO") is a glycoprotein that is the primary regulator of erythropoiesis. Specifically, EPO is responsible for promoting the growth, differentiation and survival of erythroid progenitors, which give rise to mature red blood cells. In response to changes in the level of oxygen in the blood and tissues, erythropoietin appears to stimulate both proliferation and differentiation of immature erythroblasts. It also functions as a growth factor, stimulating the mitotic activity of erythroid progenitor cells, such as erythrocyte burst forming and colony-forming units. It also acts as a differentiation factor, triggering transformation of an erythrocyte colony-forming-unit into a proerythroblast (See Erslev, A., New Eng. J. Med., 316:101-103 (1987)).

[0004] EPO has a molecular weight of about 34,000 daltons and can occur in three forms--alpha, beta and asialo. During mid- to late gestation, EPO is synthesized in the fetal liver. Subsequently, EPO is synthesized in the kidney, circulates in the plasma and is excreted in the urine.

[0005] Human urinary EPO has been isolated and purified (See, Miyake et al., J. Biol. Chem., 252:5558 (1977)). Moreover, methods for identifying, cloning and expressing genes encoding EPO (See U.S. Pat. No. 4,703,008) as well as purifying recombinant EPO from a cell medium (See U.S. Pat. No. 4,667,016) are known in the art.

[0006] The activity of EPO is mediated through the binding and activation of a cell surface receptor referred to as the erythropoietin receptor. The EPO receptor belongs to the cytokine receptor superfamily and is believed to contain at least two distinct polypeptides, a 55-72 kDa species and a 85-100 kDa species (See U.S. Pat. No. 6,319,499, Mayeux et al., J. Biol. Chem., 266:23380 (1991), McCaffery et al., J. Biol. Chem., 264:10507 (1991)). Other studies have revealed other polypeptide complexes of EPO receptor having molecular weights such as 110, 130 and 145 kDa (See U.S. Pat. No. 6,319,499).

[0007] Both the murine and human EPO receptors have been cloned and expressed (See D'Andrea et al., Cell, 57:277 (1989); Jones et al., Blood, 76:31 (1990); Winkelmann et al., Blood, 76:24 (1990); WO 90/08822/U.S. Pat. No. 5,278,065). The full length human EPO receptor is a 483 amino acid transmembrane protein with an approximately 25 amino acid signal peptide (See U.S. Pat. No. 6,319,499). The human receptor demonstrates about a 82% amino acid sequence homology with the murine receptor. Id.

[0008] In the absence of ligand the EPO receptor exists in a preformed dimer. The binding of EPO to its receptor causes a conformational change such that the cytoplasmic domains are placed in close proximity. While not completely understood, it is believed that this "dimerization" plays a role in the activation of the receptor. The activation of the EPO receptor results in a number of biological effects. Some of these activities include stimulation of proliferation, stimulation of differentiation and inhibition of apoptosis (See U.S. Pat. No. 6,319,499, Liboi et al., PNAS USA, 90:11351 (1993), Koury, Science, 248:378 (1990)).

[0009] It is the relationship between the EPO receptor dimerization and activation that can be used to identify compounds (i.e. such as antibodies) other than EPO that are capable of: (1) dimerizing the EPO receptor; and (2) activating the receptor. These compounds would be useful in treating mammals suffering from anemia and in identifying mammals having a dysfunctional EPO receptor.

SUMMARY OF THE INVENTION

[0010] In one embodiment, the invention relates to antibodies that bind to the human erythropoietin receptor. In one embodiment, the antibodies comprise a heavy chain variable region that is selected from the group consisting of SEQ ID NOS: 3, 7, 11, 15, 19, 31, 35, 39, 43, 47, 51, 55 and fragments thereof. In another embodiment, the antibodies comprise a light chain variable region that is selected from the group consisting of SEQ ID NOS: 5, 9, 13, 17, 21, 23, 25, 27, 29, 33, 37, 41, 45, 49, 53, 57 and fragments thereof.

[0011] In another embodiment, the present invention relates to an isolated antibody that is capable of binding a human erythropoietin receptor in a mammal. Such an antibody comprises a heavy chain variable region or a light chain variable region that comprises a continuous sequence from CDR1 through CDR3. The amino acid sequence of the heavy chain variable region comprising the continuous sequence from CDR1 through CDR3 is selected from the group consisting of: SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61 and fragments thereof. The amino acid sequence of the light chain variable region comprising the continuous sequence from CDR1 through CDR3 is selected from the group consisting of: SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68 and fragments thereof.

[0012] In another embodiment, the present invention relates to an antibody that activates an endogenous activity of a human erythropoietin receptor in a mammal but does not interact with a peptide having an amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO:1).

[0013] In another embodiment, the present invention relates to an antibody that is capable of activating an endogenous activity of a human erythropoietin receptor in a mammal, wherein said antibody or antibody fragment thereof exhibits a binding affinity within one hundred fold of the binding affinity of endogenous human erythropoietin to the erythropoietin receptor.

[0014] In yet another embodiment, the present invention relates to an antibody or antibody fragment thereof that activates an endogenous activity of a human erythropoietin receptor in a mammal. The antibody or antibody fragment thereof comprises at least one human heavy chain variable region having the amino acid sequence of SEQ ID NO:3 or antibody fragment thereof, and/or at least one human light chain variable region having the amino acid sequence of SEQ ID NO:5 or antibody fragment thereof, provided that said antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO: 1).

[0015] In yet another embodiment, the present invention relates to an antibody or antibody fragment thereof that activates an endogenous activity of a human erythropoietin receptor in a mammal. The antibody or antibody fragment thereof comprises at least one heavy chain variable region having the amino acid sequence of SEQ ID NO:7 or antibody fragment thereof, and/or at least one light chain variable region having the amino acid sequence of SEQ ID NO:9 or antibody fragment thereof, provided that said antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO:1).

[0016] This embodiment also includes other heavy chain variable regions selected from the group consisting of SEQ ID NO: 11, 15, 19, 31, 35, 39, 43, 47, 51, and 55 or an antibody fragment of any of these aforementioned SEQ ID NOS, wherein said antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of SEQ ID NO: 1. Other light chain variable regions included in this embodiment may be selected from the group consisting of SEQ ID NO: 13, 17, 21, 23, 25, 27, 29, 33, 37, 41, 45, 49, 53 and 57 or an antibody fragment of any of these aforementioned SEQ ID NOS, wherein said antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of SEQ ID NO:1.

[0017] In yet another embodiment, the invention provides an antibody or antibody fragment thereof that activates an endogenous activity of a human erythropoietin receptor in a mammal, the antibody comprising the amino acid sequences of at least one heavy chain variable region and at least one light chain variable region selected from the group consisting of SEQ ID NO:11/SEQ ID NO:13, SEQ ID NO:15/SEQ ID NO:17, SEQ ID NO: 19/SEQ ID NO:21, SEQ ID NO:11/SEQ ID NO:23, SEQ ID NO:11/SEQ ID NO:25, SEQ ID NO:11/SEQ ID NO:27, SEQ ID NO:11/SEQ ID NO:29, SEQ ID NO:31/SEQ ID NO:33, SEQ ID NO:35/SEQ ID NO:37, SEQ ID NO:39/SEQ ID NO:41, SEQ ID NO:43/SEQ ID NO:45, SEQ ID NO:47/SEQ ID NO:49, SEQ ID NO:51/SEQ ID NO:53 and SEQ ID NO:55/SEQ ID NO:57 or antibody fragment thereof, wherein said antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of SEQ ID NO: 1.

[0018] In yet another embodiment, the present invention relates to a method of activating an endogenous activity of a human erythropoietin receptor in a mammal. The method involves the step of administering to a mammal a therapeutically effective amount of an antibody or antibody fragment thereof to activate the EPO receptor. The antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO:1).

[0019] In yet a further embodiment, the present invention relates to a method of modulating an endogenous activity of a human erythropoietin receptor in a mammal. The method involves administering to a mammal a therapeutically effective amount of an antibody or antibody fragment thereof to modulate the endogenous activity of a human erythropoietin receptor in a mammal but does not interact with a peptide having an amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO: 1).

[0020] In yet a further embodiment, the present invention relates to a method of treating a mammal suffering from pure red cell aplasia induced by neutralizing anti-erythropoietin antibodies. The method involves administering to a mammal in need of treatment a therapeutically effective amount of an antibody or antibody fragment thereof to activate said receptor, wherein said antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO:1).

[0021] In yet a further embodiment, the present invention relates to pharmaceutical compositions. The pharmaceutical compositions of the present invention contain a therapeutically effective amount of an antibody or antibody fragment thereof and a pharmaceutically acceptable excipient. The antibody or antibody fragment contained in the pharmaceutical composition activates an endogenous activity of a human erythropoietin receptor in a mammal but does not interact with a peptide having an amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO:1).

[0022] In yet a further embodiment, the present invention relates to an IgG2 antibody or antibody fragment that binds to and activates the erythropoietin receptor. The IgG2 antibodies or antibody fragments of this embodiment bind to and interact with any epitope that is involved in activating the EPO receptor. Such antibodies may be polyclonal or monoclonal antibodies or any antibody fragment thereof. The IgG2 antibodies may be chimeric, humanized or human antibodies.

[0023] In yet a further embodiment, the present invention provides a method of activating an endogenous activity of a human erythropoietin receptor in a mammal comprising the step of administering to a mammal a therapeutically effective amount of an IgG2 antibody or antibody fragment of the invention to activate the receptor.

[0024] In yet a further embodiment, the present invention provides a method of modulating an endogenous activity of a human erythropoietin receptor in a mammal comprising the step of administering to a mammal a therapeutically effective amount of an IgG2 antibody or antibody fragment of the invention to modulate the receptor.

[0025] In yet another embodiment, the present invention provides a method of treating a mammal suffering aplasia, the method comprising the step of administering to a mammal in need of treatment a therapeutically effective amount of an IgG2 antibody or antibody fragment of the invention to activate the receptor.

[0026] In yet another embodiment, the present invention provides a method of treating a mammal suffering aplasia, the method comprising the step of administering to a mammal in need of treatment a therapeutically effective amount of an IgG2 antibody or antibody fragment of the invention to modulate the receptor.

[0027] In yet another embodiment, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of an IgG2 antibody or antibody fragment of the invention and a pharmaceutically acceptable excipient.

[0028] Finally, the present invention relates to isolated and purified polynucleotide and amino acid sequences. The isolated and purified polynucleotide sequences can be selected from the group consisting of: SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO: 18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, SEQ ID NO:42, SEQ ID NO:44, SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, SEQ ID NO:56 and fragments, complements and degenerate codon equivalents thereof.

[0029] The present invention further relates to isolated and purified amino acid sequences selected from the group consisting of: SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:25, SEQ ID NO:29, SEQ ID NO:311, SEQ ID NO:33, SEQ ID NO:35, SEQ ID NO:37, SEQ ID NO:39, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53. SEQ ID NO:55, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68 and fragments and complements and thereof.

BRIEF DESCRIPTION OF THE FIGURES

[0030] FIG. 1 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:69 and SEQ ID NO:70, respectively) and amino acid sequence of the heavy chain of human antibody Abl 2. The amino acid sequence comprises SEQ ID NOS:71 through 74. The sequence of the constant region alone is shown as SEQ ID NO:75. The variable chain ends at nucleotide 1283. The variable/constant joining region (underlined) is at nucleotides 1284-1289. The constant region is from nucleotides 1290-2826.

[0031] FIG. 2 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:76 and SEQ ID NO:77, respectively) and amino acid sequence of the light chain of human antibody Ab12. The amino acid sequence comprises SEQ ID NOS:78. The sequence of the constant region alone is shown as SEQ ID NO: 79. The variable chain ends at nucleotide 1363. The variable/constant joining region (underlined) is at nucleotides 1364-1369. The constant region is from nucleotides 1370-1618.

[0032] FIG. 3 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:80 and SEQ ID NO:81, respectively) and amino acid sequence of the heavy chain of human antibody Ab198. The amino acid sequence comprises SEQ ID NOS:82 and SEQ ID NOS 72 through 74. The variable chain ends at nucleotide 1304. The variable/constant joining region (underlined) is at nucleotides 1305-1310. The constant region is from nucleotides 1311-2847.

[0033] FIG. 4 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:83 and SEQ ID NO:84, respectively) and amino acid sequence of the light chain of human antibody Ab198. The amino acid sequence comprises SEQ ID NOS:78. The variable chain ends at nucleotide 1351. The variable/constant joining region (underlined) is at nucleotides 1352-1357. The constant region is from nucleotides 1358-1606.

[0034] FIG. 5 shows the competition of Ab12 with .sup.125I-labeled EPO for binding to Chinese Hamster Ovary cells expressing recombinant EPO receptor.

[0035] FIG. 6 shows the results of an EPO dependent human cell proliferation assay using Ab12 and Ab198.

[0036] FIG. 7 shows that Ab12 remains active in inducing the proliferation of F36E cells after storage at 4.degree. C. for up to 20 days.

[0037] FIG. 8 shows that Ab12 induces the formation of CFU-E (colony forming unit-erythroid) from human 36.sup.+ progenitor cells.

[0038] FIG. 9 shows the induction of proliferation of human erythroid producing cells with Ab198.

[0039] FIG. 10 shows that Ab198 induces the formation of CFU-E colonies from cynomologous bone marrow-derived erythroid progenitor cells.

[0040] FIG. 11 shows that Ab12 does not interact with the peptide SE-3. Ab71A interacts with the SE-3 peptide.

[0041] FIG. 12 shows that human Abs secreted by primary hybridomas induce the proliferation of F36E cells.

[0042] FIG. 13 shows that human Ab supernatants secreted by primary hybridomas interact with intact EPO receptor, but not with peptide SE-3.

[0043] FIG. 14 shows the activity of various concentrations of Ab12 on the proliferation of UT7/EPO cells.

[0044] FIG. 15 shows the activity of various concentrations of Ab 198 on the proliferation of UT7/EPO cells.

[0045] FIG. 16 shows the activity of various concentrations of Abl 98 (with or without the addition of a secondary goat anti-human FC antibody) on the growth and proliferation of UT7/EPO cells.

[0046] FIG. 17 shows the activity of various concentrations of Ab12 (with or without the addition of a secondary goat anti-human FC antibody) on the growth and proliferation of UT7/EPO cells.

[0047] FIG. 18 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-003 of the invention, with FIG. 18A (SEQ ID NO:10) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 18B (SEQ ID NO:11) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 18A, FIG. 18C (SEQ ID NO: 12) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 18D (SEQ ID NO:13) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 18C.

[0048] FIG. 19 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-012 (also referred to herein as Abl 2) of the invention, with FIG. 19A (SEQ ID NO:2) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 19B (SEQ ID NO:3) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 19A, FIG. 19C (SEQ ID NO:4) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 19D (SEQ ID NO:5) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 19C.

[0049] FIG. 20 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-022 of the invention, with FIG. 20A (SEQ ID NO: 14) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 20B (SEQ ID NO:15) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 20A, FIG. 20C (SEQ ID NO:16) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 20D (SEQ ID NO: 17) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 20C.

[0050] FIG. 21 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-054 of the invention, with FIG. 21A (SEQ ID NO:18) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 21B (SEQ ID NO:19) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 21A, FIG. 21C (SEQ ID NO:20) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 21D (SEQ ID NO:21) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 21C.

[0051] FIG. 22 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-060 of the invention, with FIG. 22A (SEQ ID NO: 10) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 22B (SEQ ID NO:11) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 22A. FIG. 22C (SEQ ID NO:22) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 22D (SEQ ID NO:23) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 22C.

[0052] FIG. 23 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-102 of the invention, with FIG. 23A (SEQ ID NO: 10) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 23B (SEQ ID NO: 11) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 23A, FIG. 23C (SEQ ID NO:24) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 23D (SEQ ID NO:25) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 23C.

[0053] FIG. 24 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-135 of the invention, with FIG. 24A (SEQ ID NO: 10) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 24B (SEQ ID NO: 11) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 24A, FIG. 24C (SEQ ID NO:26) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 24D (SEQ ID NO:27) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 24C.

[0054] FIG. 25 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-145 of the invention. with FIG. 25A (SEQ ID NO:10) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 25B (SEQ ID NO:11) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 25A, FIG. 25C (SEQ ID NO:28) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 25D (SEQ ID NO:29) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 25C.

[0055] FIG. 26 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-198 (also referred to herein as Ab198) of the invention, with FIG. 26A (SEQ ID NO:6) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 26B (SEQ ID NO:7) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 26A, FIG. 26C (SEQ ID NO:8) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 26D (SEQ ID NO:9) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 26C.

[0056] FIG. 27 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-254 of the invention, with FIG. 27A (SEQ ID NO:30) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 27B (SEQ ID NO:31) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 27A, FIG. 27C (SEQ ID NO:32) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 27D (SEQ ID NO:33) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 27C.

[0057] FIG. 28 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-267 of the invention, with FIG. 28A (SEQ ID NO:34) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 28B (SEQ ID NO:35) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 28A. FIG. 28C (SEQ ID NO:36) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 28D (SEQ ID NO:37) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 28C.

[0058] FIG. 29 is a table showing amino acid sequence alignments of heavy chain variable regions of anti-EPOr mAbs generated according to the invention with their associated germline variable region sequences and identifying framework regions and complementarity determining regions.

[0059] FIG. 30 is a table showing amino acid sequence alignments of light chain variable regions of anti-EPOr mAbs generated according to the invention with their associated germline variable region sequences and identifying framework regions and complementarity determining regions.

[0060] FIG. 31 is a graph comparing the erythropoietic activity, at various concentrations, of a gamma-1 Ab 12 monoclonal antibody (Mab) and a gamma-2 Ab 12 Mab on an F36e human erythroleukemic cell line.

[0061] FIG. 32 is a graph showing the increase in percent reticulocyte and percent hematocrit in transgenic mice subjected to a multiple dosing regimen of vehicle, Epogen (5 U) or Ab 12 antibody (5 or 50 .mu.g).

[0062] FIG. 33 is a graph showing the increase in percent hematocrit in transgenic mice subjected to a weekly dosing regimen (over 3 weeks) of various concentrations of Aranesp.TM. or Ab 12.

[0063] FIG. 34 is a graph showing the increase in percent hematocrit in transgenic mice subjected to single versus weekly dosing regimens of various concentrations of Aranesp.TM. or Ab 12.

[0064] FIG. 35 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-390 of the invention, with FIG. 35A (SEQ ID NO:38) representing the nucleotide sequence encoding the variable region of the heavy chain. FIG. 35B (SEQ ID NO:39) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 35A, FIG. 35C (SEQ ID NO:40) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 35D (SEQ ID NO:41) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 35C.

[0065] FIG. 36 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-412 of the invention, with FIG. 36A (SEQ ID NO:42) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 36B (SEQ ID NO:43) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 36A, FIG. 36C (SEQ ID NO:44) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 36D (SEQ ID NO:45) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 36C.

[0066] FIG. 37 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-430/432 of the invention, with FIG. 37A (SEQ ID NO:46) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 37B (SEQ ID NO:47) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 37A, FIG. 37C (SEQ ID NO:48) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 37D (SEQ ID NO:49) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 37C.

[0067] FIG. 38 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-467 of the invention, with FIG. 38A (SEQ ID NO:50) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG. 38B (SEQ ID NO:51) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 38A, FIG. 38C (SEQ ID NO:52) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 38D (SEQ ID NO:53) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 38C.

[0068] FIG. 39 is a series of representations of the heavy chain and light chain variable region nucleotide and amino acid sequences of the human anti-EPO-R antibody expressed by the cell line designated ABT2-SCX-484 of the invention, with FIG. 39A (SEQ ID NO:54) representing the nucleotide sequence encoding the variable region of the heavy chain, FIG., 39B (SEQ ID NO:55) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 39A, FIG. 39C (SEQ ID NO:56) representing the nucleotide sequence encoding the variable region of the light chain, and FIG. 39D (SEQ ID NO:57) representing the amino acid sequence encoded by the nucleotide sequence shown in FIG. 39C.

[0069] FIG. 40 is a table showing amino acid sequence alignments of heavy chain variable regions of anti-EPOr mAbs generated according to the invention with their associated germline variable region sequences and identifying framework regions and complementarity determining regions.

[0070] FIG. 41 is a table showing amino acid sequence alignments of light chain variable regions of anti-EPOr mAbs generated according to the invention with their associated germline variable region sequences and identifying framework regions and complementarity determining regions.

[0071] FIG. 42 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:86 and SEQ ID NO:87, respectively) and amino acid sequence of the heavy chain of human antibody Ab390. The amino acid sequence comprises SEQ ID NOS:88 and SEQ ID NOS 72 through 74. The variable chain ends at nucleotide 463. The variable/constant joining region (underlined) is at nucleotides 464-469. The constant region is from nucleotides 470-2006.

[0072] FIG. 43 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:89 and SEQ ID NO:90, respectively) and amino acid sequence of the light chain of human antibody Ab390. The amino acid sequence comprises SEQ ID NOS:91. The variable chain ends at nucleotide 463. The variable/constant joining region (underlined) is at nucleotides 464-469. The constant region is from nucleotides 470-718.

[0073] FIG. 44 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:92 and SEQ ID NO:93, respectively) and amino acid sequence of the heavy chain of human antibody Ab412. The amino acid sequence comprises SEQ ID NOS:94 and SEQ ID NOS 72 through 74. The variable chain ends at nucleotide 469. The variable/constant joining region (underlined) is at nucleotides 470-475. The constant region is from nucleotides 476-2012.

[0074] FIG. 45 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:95 and SEQ ID NO:96, respectively) and amino acid sequence of the light chain of human antibody Ab412. The amino acid sequence comprises SEQ ID NOS:97. The variable chain ends at nucleotide 463. The variable/constant joining region (underlined) is at nucleotides 464-469. The constant region is from nucleotides 470-718.

[0075] FIG. 46 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:98 and SEQ ID NO:99, respectively) and amino acid sequence of the heavy chain of human antibody Ab432. The amino acid sequence comprises SEQ ID NOS: 100 and SEQ ID NOS 72 through 74. The variable chain ends at nucleotide 463. The variable/constant joining region (underlined) is at nucleotides 464-469. The constant region is from nucleotides 470-2006.

[0076] FIG. 47 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:101 and SEQ ID NO:102, respectively) and amino acid sequence of the light chain of human antibody Ab430. The amino acid sequence comprises SEQ ID NOS:103. The variable chain ends at nucleotide 463. The variable/constant joining region (underlined) is at nucleotides 464-469. The constant region is from nucleotides 470-718.

[0077] FIG. 48 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:104 and SEQ ID NO:105, respectively) and amino acid sequence of the heavy chain of human antibody Ab467. The amino acid sequence comprises SEQ ID NOS:106 and SEQ ID NOS 72 through 74. The variable chain ends at nucleotide 463. The variable/constant joining region (underlined) is at nucleotides 464-469. The constant region is from nucleotides 470-2006.

[0078] FIG. 49 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:107 and SEQ ID NO:108, respectively) and amino acid sequence of the light chain of human antibody Ab467. The amino acid sequence comprises SEQ ID NOS:109. The variable chain ends at nucleotide 463. The variable/constant joining region (underlined) is at nucleotides 464-469. The constant region is from nucleotides 470-718.

[0079] FIG. 50 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:110 and SEQ ID NO:111, respectively) and amino acid sequence of the heavy chain of human antibody Ab484. The amino acid sequence comprises SEQ ID NOS:112 and SEQ ID NOS 72 through 74. The variable chain ends at nucleotide 469. The variable/constant joining region (underlined) is at nucleotides 470-475. The constant region is from nucleotides 470-2012.

[0080] FIG. 51 shows the isolated and purified polynucleotide (top strand and bottom strands, SEQ ID NO:113 and SEQ ID NO:114, respectively) and amino acid sequence of the light chain of human antibody Ab484. The amino acid sequence comprises SEQ ID NOS:115. The variable chain ends at nucleotide 463. The variable/constant joining region (underlined) is at nucleotides 464-469. The constant region is from nucleotides 470-718.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

[0081] As used herein, the term "antibody" or "immunoglobulin" refers to single chain, two-chain, and multi-chain proteins and glycoproteins that belong to the classes of polyclonal, monoclonal, chimeric and human or humanized. The term "antibody" also includes synthetic and genetically engineered variants thereof.

[0082] As used herein, the term "antibody fragment" refers to Fab, Fab', F(ab').sub.2 and Fv fragments, as well as any portion of an antibody having specificity toward at least one desired epitope.

[0083] As used herein, the term "gamma-2", "gamma-2 isotype" or "IgG2" refers to subclass 2 of immunoglobulin G (IgG), as well as any antibody fragment thereof. The four subclasses of IgG molecules are well characterized and well known to those of ordinary skill in the art. (See, for example, Molecular Biology of the Cell, 2.sup.nd Edition by Bruce Alberts et al., 1989) Panels of monoclonal antibodies are available that recognize all human isotypes (IgA, IgG, IgD IgE, and IgM) and subisotypes (IgA1, IgA2, IgG1, IgG2, IgG3, and IgG4) of human immunoglobulins.

[0084] As used herein the term "humanized antibody" refers to an antibody that is derived from a non-human antibody (i.e murine) that retains or substantially retains the antigen-binding properties of the parent antibody but is less immunogenic in humans.

[0085] As used herein, the term "human antibody" refers to an antibody that possesses a sequence that is derived from a human germ-line immunoglobulin sequence, such as antibodies derived from transgenic mice having human immunoglobulin genes (e.g., XenoMouse.RTM. mice), human phage display libraries, or human B cells.

[0086] As used herein, the term "epitope" refers to any protein determinate capable of specifically binding to an antibody or T-cell receptors. Epitopic determinants usually consist of chemically active surface groupings of molecules such as amino acids or sugar side chains and usually have specific three dimensional structural characteristics, as well as specific charge characteristics.

[0087] As used herein, the term "endogenous" refers to a product or activity arising in the body or cell as opposed to a product or activity coming from outside.

[0088] As used herein the phrase, a polynucleotide "derived from" or "specific for a designated sequence refers to a polynucleotide sequence that comprises a contiguous sequence of approximately at least 6 nucleotides, preferably at least about 8 nucleotides, more preferably at least about 10-12 nucleotides, and even more preferably at least about 15-20 nucleotides corresponding, i.e., identical or complementary to, a region of the designated nucleotide sequence. The sequence may be complementary or identical to a sequence that is unique to a particular polynucleotide sequence as determined by techniques known in the art. Regions from which sequences may be derived, include but are not limited to, regions encoding specific epitopes, as well as non-translated and/or non-transcribed regions.

[0089] The derived polynucleotide will not necessarily be derived physically from the nucleotide sequence of interest under study, but may be generated in any manner, including, but not limited to, chemical synthesis, replication, reverse transcription or transcription, that is based on the information provided by the sequence of bases in the region(s) from which the polynucleotide is derived. As such, it may represent either a sense or an antisense orientation of the original polynucleotide. In addition, combinations of regions corresponding to that of the designated sequence may be modified in ways known in the art to be consistent with the intended use.

[0090] As used herein, the phrase "encoded by" refers to a nucleic acid sequence that codes for a polypeptide sequence, wherein the polypeptide sequence or a portion thereof contains an amino acid sequence of at least 3 to 5 amino acids, more preferably at least 8 to 10 amino acids, and even more preferably at least 15 to 20 amino acids from a polypeptide encoded by the nucleic acid sequence. Also encompassed are polypeptide sequences that are immunologically identifiable with a polypeptide encoded by the sequence. Thus, a "polypeptide," "protein" or "amino acid" sequence has at least about 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% or more identity to the antibodies of the present invention. Further, the antibodies of the present invention may have at least about 60%, 70%, 75%, 80%, 85%, 90% or 95% similarity to a polypeptide or amino sequences of the antibodies of the present invention. The amino acid sequences of the antibodies of the present invention can be selected from the group consisting of SEQUENCE ID NOS: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55 and 57. Preferred amino acid sequences of the antibodies of the present invention are selected from the group consisting of SEQ ID NOS: 3, 5, 7, 9, 51 and 53.

[0091] As used herein, the phrase "recombinant polypeptide," "recombinant protein," or "a polypeptide produced by recombinant techniques", which terms may be used interchangeably herein, describes a polypeptide that by virtue of its origin or manipulation is not associated with all or a portion of the polypeptide with which it is associated in nature and/or is linked to a polypeptide other than that to which it is lined in nature. A recombinant or encoded polypeptide or protein is not necessarily translated from a designated nucleic acid sequence. It also may be generated in any manner, including chemical synthesis or expression of a recombinant expression system.

[0092] As used herein, the phrase "synthetic peptide" refers to a polymeric form of amino acids of any length, which may be chemically synthesized by methods well known in the art (See U.S. Pat. Nos. 4,816,513, 5,854,389, 5,891,993 and 6,184,344).

[0093] As used herein, the term "polynucleotide" refers to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. This term refers only to the primary structure of the molecule. Thus, the term includes double and single-stranded DNA as well as double- and single-stranded RNA. It also includes modifications, such as methylation or capping and unmodified forms of the polynucleotide. The terms "polynucleotide", "oligomer," "oligonucleotide," and "oligo," are used interchangeably herein.

[0094] As used herein the phrase "purified polynucleotide" refers to a polynucleotide of interest or fragment thereof that is essentially free, e.g. contains less than about 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% of the protein with which the polynucleotide is naturally associated. Techniques for purifying polynucleotides of interest are well known in the art and include, for example, disruption of the cell containing the polynucleotide with a chaotropic agent and separation of the polynucleotide(s) and proteins by ion-exchange chromatography, affinity chromatography and sedimentation according to density.

[0095] As used herein, the phrase "purified polypeptide" or "purified protein" means a polypeptide of interest or fragment thereof which is essentially free of, e.g., contains less than about 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, cellular components with which the polypeptide of interest is naturally associated. Methods for purifying polypeptides of interest are known in the art.

[0096] As used herein, the term "isolated" refers to material that is removed from its original environment (e.g., the natural environment if it is naturally occurring). For example, a naturally occurring polynucleotide or polypeptide present in a living animal is not isolated, but the same polynucleotide or DNA or polypeptide, separated from some or all of the coexisting materials in the natural system, is isolated. Such polynucleotide could be part of a vector and/or such polynucleotide or polypeptide could be part of a composition, and still be isolated in that the vector or composition is not part of its natural environment.

[0097] As used herein, the term "polypeptide" and "protein" are used interchangeably and refer to at least one molecular chain of amino acids linked through covalent and/or non-covalent bonds. The terms do not refer to a specific length of the product. Thus, peptides, oligopeptides and proteins are included within the definition of polypeptide. The terms include post-translational modifications of the polypeptide, including, but not limited to, glycosylations, acetylations, phosphorylations and the like. In addition, protein fragments, analogs, mutated or variant proteins, fusion proteins and the like are included within the meaning of polypeptide.

[0098] As used herein, the phrase "recombinant host cells," "host cells," "cells," "cell lines," "cell cultures," and other such terms denoting microorganisms or higher eukaryotic cell lines cultured as unicellular entities refer to cells that can be, or have been, used as recipients for recombinant vector or other transferred DNA, and include the original progeny of the original cell that has been transfected.

[0099] As used herein, the term "replicon" refers to any genetic element, such as a plasmid, a chromosome or a virus, that behaves as an autonomous unit of polynucleotide replication within a cell.

[0100] As used herein, the term "operably linked" refers to a situation wherein the components described are in a relationship permitting them to function in their intended manner. Thus, for example, a control sequence "operably linked" to a coding sequence is ligated in such a manner that expression of the coding sequence is achieved under conditions compatible with the control sequence.

[0101] As used herein, the term "vector" refers to a replicon in which another polynucleotide segment is attached, such as to bring about the replication and/or expression of the attached segment.

[0102] As used herein, the term "control sequence" refers to a polynucleotide sequence that is necessary to effect the expression of a coding sequence to which it is ligated. The nature of such control sequences differs depending upon the host organism. In prokaryotes, such control sequences generally include a promoter, a ribosomal binding site and terminators and, in some instances, enhancers. The term "control sequence" thus is intended to include at a minimum all components whose presence is necessary for expression, and also may include additional components whose presence is advantageous, for example, leader sequences.

[0103] The term "transfection" refers to the introduction of an exogenous polynucleotide into a prokaryotic or eucaryotic host cell, irrespective of the method used for the introduction. The term "transfection" refers to both stable and transient introduction of the polynucleotide, and encompasses direct uptake of polynucleotides, transformation, transduction and f-mating. Once introduced into the host cell, the exogenous polynucleotide may be maintained as a non-integrated replicon, for example, a plasmid, or alternatively, may be integrated into the host genome.

[0104] As used herein, the term "treatment" refers to prophylaxis and/or therapy.

[0105] As used herein, the term "purified product" refers to a preparation of the product which has been isolated from the cellular constituents with which the product is normally associated and from other types of cells that may be present in the sample of interest.

[0106] As used herein, the phrase "activation of an erythropoietin (EPO) receptor" refers to one or more molecular processes which an EPO receptor undergoes that result in the transduction of a signal to the interior of a receptor-bearing cell. Ultimately, this signal brings about one or more changes in cellular physiology. Activation of the EPO receptor typically results in the proliferation or differentiation of EPO receptor-bearing cells, such as, but not limited to, erythroid progenitor cells. A number of events are involved in the activation of the EPO receptor, such as, but not limited to, the dimerization of the receptor.

[0107] The structural unit of an antibody is a tetramer. Each tetramer is composed of two identical pairs of polypeptide chains, each pair having one "light" (25 kDa) and one "heavy" chain (about 50-70 kDa). The amino-terminal portion of each chain includes a variable region that is primarily responsible for antigen recognition. The carboxy-terminal portion of the chain defines a constant region that is responsible for the effector function of the antibody. Human light chains are classified as kappa and lambda light chains. Heavy chains are classified as mu, delta, gammna, alpha, or epsilon and define the antibody's isotype as IgM, IgD, IgG, IgA, and IgE. IgG immunoglobulins are classified further into four subclasses (IgG1, IgG2, IgG3 and IgG4) having gamma-1, gamma-2, gamma-3 and gamma-4 heavy chains, respectively. Most of the therapeutic human, chimeric or humanized antibodies available are of the IgG1 antibody type including Herceptin for breast cancer, Rituxan for Non-Hodgkins lymphoma and Humira and Remicade for rheumatoid arthritis (See Glennie, M. J. et al. Drug Discovery Today, 8:503 (2003).

[0108] Within the light and heavy chains, the variable and constant regions are joined by a "J" region with the heavy chain also include a "D" region. The variable regions of each light/heavy chain pair form the antigen binding site. Thereupon, an intact antibody has two binding sites, which, except in bifunctional or bispecific antibodies, are the same. Bifunctional or bispecific antibodies are artificial hybrid antibodies that have two different heavy/light chain pairs and two different binding sites. Bifunctional or bispecific antibodies can be produced using routine techniques known in the art.

[0109] The structure of the chains of an antibody exhibit the same general structure of relatively conserved framework regions (FR) joined by three hyper variable regions, also called complementarity determining regions or CDRs. The CDRs from the two chains of each pair are aligned by the framework regions, enabling binding to a specific epitope. From N-terminal to C-terminal, both the light and heavy chains comprise the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4.

[0110] U.S. Pat. No. 6,319,499 describes antibodies that bind to and activate an erythropoietin receptor (EPO-R). The antibodies specifically identified in this patent are Mabs 71 and 73. Mab 71 binds to a peptide designated "SE-3" having the amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO:1) (See Example 3). SE-3 is located on the human EPO-R between amino acid residues 49-78. According to U.S. Pat. No. 6,319,499, when this region of the EPO-R (i.e. amino acid residues 49-79) is bound with a cross linker such as Mab 71, this results in the activation of the EPO receptor. Example 6 in U.S. Pat. No. 6,319,499 states that Mab 71 binds "significant amounts of peptide SE-3" compared to other peptides tested. This example further states that this "indicates that Mab 71 binds to a region of the human EPO-R containing or overlapping residues 49 to 78." Mabs 71 and 73 are murine antibodies. Although rodent and human antibodies may both provide precision for target specificity, human antibodies interact far more effectively with the natural defenses of the body and do not elicit anti-antibody responses to the same extent as rodent antibodies (Winter, G. and Milstein, C. Nature 349: 293 (1991). Additionally, the flexibility of human IgG subclasses differ (Roux, K. H. et al., J Immunol. 159: 3372 (1997) and this difference also extends to rodent IgG isotypes since rodent IgG isotypes differ from their human counterparts. Since protein flexibility may affect antibody-antigen recognition (Jimenez, R., et al. Proc. Natl. Acad. Sci. USA, 100: 92 (2003), human IgG2 isotypes may result in antigen recognition mechanisms distinct from those of murine antibodies. Murine IgG isotypes generally differ from those of humans.

[0111] In one embodiment, the present invention relates to an antibody or antibody fragment that binds to the erythropoietin receptor. The antibody or antibody fragment that binds to the erythropoietin receptor comprises at least one heavy chain having an amino acid sequence selected from the group consisting of: SEQ ID NOS: 3, 7, 11, 15, 19, 31, 35, 39, 43, 47, 51, 55 and fragments thereof. In a second embodiment, the antibody or antibody fragment that binds to the erythropoietin receptor comprises at least one light chain having an amino acid sequence selected from the group consisting of: SEQ ID NOS: 5, 9, 13, 17, 21, 23, 25, 27, 29, 33, 37, 41, 45, 49, 53, 57 and fragments thereof.

[0112] In a third embodiment, the present invention relates to an isolated antibody that is capable of binding a human erythropoietin receptor in a mammal. More specifically, the antibody comprises a heavy chain variable region or a light chain variable region which comprises a continuous sequence from CDR1 through CDR3. The amino acid sequence of the heavy chain variable region comprising the continuous sequence from CDR1 through CDR3 is selected from the group consisting of: SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60 and SEQ ID NO:61, and fragments thereof. The amino acid sequence of the light chain variable region comprising the continuous sequence from CDR1 through CDR3 is selected from the group consisting of: SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, and SEQ ID NO:68, and fragments thereof. In addition, the present invention relates to an isolated antibody which comprises a heavy chain variable region or a light chain variable region which comprises at least one CDR. More specifically, the antibody comprises a heavy chain variable region comprising at least one CDR selected from the group consisting of amino acid residues 99-112 of SEQ ID NO:11, 26-35 of SEQ ID NO:3, 50-65 of SEQ ID NO:3, 98-105 of SEQ ID NO:3, 26-35 of SEQ ID NO:19, 50-66 of SEQ ID NO:19.99-105 of SEQ ID NO:19, 50-66 of SEQ ID NO:31, 99-105 of SEQ ID NO:31, 26-35 of SEQ ID NO:39, 50-65 of SEQ ID NO:39, 98-105 of SEQ ID NO:39, 26-37 of SEQ ID NO:43, 52-67 of SEQ ID NO:43, 100-107 of SEQ ID NO:43, 26-35 of SEQ ID NO:47, 50-65 of SEQ ID NO:47, 26-35 of SEQ ID NO:5, 50-65 of SEQ ID NO:51, 98-105 of SEQ ID NO:51, 26-37 of SEQ ID NO:55 and 52-67 of SEQ ID NO:55 or a light chain variable region comprising at least one CDR selected from the group consisting of amino acid residues 24-34 of SEQ ID NO:13, 50-56 of SEQ ID NO:13, 89-97 of SEQ ID NO:5, 24-34 of SEQ ID NO:27, 50-56 of SEQ ID NO:9, 24-39 of SEQ ID NO:33, 55-61 of SEQ ID NO:33, 24-34 of SEQ ID NO:41, 89-97 of SEQ ID NO:41, 24-34 of SEQ ID NO:45, 50-56 of SEQ ID NO:45, 89-97 of SEQ ID NO:45, 89-97 of SEQ ID NO:49 and 24-34 of SEQ ID NO:57.

[0113] In a fourth embodiment, the present invention relates to an antibody or antibody fragment that binds to and activates the erythropoietin receptor. The antibodies of the present invention bind to at least one epitope that is involved in activating the EPO receptor (Example 4). Unlike other antibodies or fragments known in the art that bind to and activate an erythropoietin receptor, such as the antibodies described in U.S. Pat. No. 6,319,499, the antibodies of the present invention do not interact with the peptide designated SE-3. Surprisingly, the antibodies of the present invention are erythropoietic even though the antibodies do not bind to the SE-3 peptide. Therefore, the human antibodies of the present invention interact with at least one different epitope on the human EPO receptor than the antibodies described in U.S. Pat. No. 6,319,499.

[0114] In a fifth embodiment, the present invention relates to an IgG2 antibody or antibody fragment that binds to and activates the erythropoietin receptor. The IgG2 antibodies or antibody fragments of this embodiment bind to and interact with any epitope that is involved in activating the EPO receptor.

[0115] Additionally, as demonstrated by the BIAcore results shown in Example 1, the antibodies of the present invention exhibit a binding affinity to the erythropoietin receptor within one hundred fold of the binding affinity of endogenous human erythropoietin to the erythropoietin receptor. A high (.about.1 nM) and low (.about.1 .mu.M) affinity of the EPO receptor for EPO has been reported resulting from two nonequivalent receptor binding sites on EPO (See Philo, J. S. et al., Biochemistry, 35:1681 (1996)).

[0116] The antibodies of the present invention can be polyclonal antibodies, monoclonal antibodies, chimeric antibodies (See U.S. Pat. No. 6,020,153) or human or humanized antibodies or antibody fragments thereof. Synthetic and genetically engineered variants (See U.S. Pat. No. 6,331,415) of any of the foregoing are also contemplated by the present invention. Preferably, however, the antibodies of the present invention are human or humanized antibodies. The advantage of human or humanized antibodies is that they potentially decrease or eliminate the immunogenicity of the antibody in a host recipient, thereby permitting an increase in the bioavailability and a reduction in the possibility of adverse immune reaction, thus potentially enabling multiple antibody administrations.

[0117] Humanized antibodies include chimeric or CDR-grafted antibodies. Also, human antibodies can be produced using genetically engineered strains of animals in which the antibody gene expression of the animal is suppressed and functionally replaced with human antibody gene expression.

[0118] Methods for making humanized and human antibodies are known in the art. One method for making human antibodies employs the use of transgenic animals, such as a transgenic mouse. These transgenic animals contain a substantial portion of the human antibody producing genome inserted into their own genome and the animal's own endogenous antibody production is rendered deficient in the production of antibodies. Methods for making such transgenic animals are known in the art. Such transgenic animals can be made using XenoMouse.RTM. technology or by using a "minilocus" approach. Methods for making Xenomice.TM. are described in U.S. Pat. Nos. 6,162,963, 6,150,584, 6,114,598 and 6,075,181. Methods for making transgenic animals using the "minilocus" approach are described in U.S. Pat. Nos. 5,545,807, 5,545,806 and 5,625,825. Also see International Publication No. WO93/12227.

[0119] Using the XenoMouse.RTM. technology, human antibodies can be obtained by immunizing a XenoMouse.RTM. mouse (Abgenix. Fremont, Calif.) with an antigen of interest. The lymphatic cells (such as B-cells) are recovered from the mice that express antibodies. These recovered cells can be fused with myeloid-type cell line to prepare immortal hybridoma cell lines. These hybridoma cell lines can be screened and selected to identify hybridoma cell lines that produce antibodies specific to the antigen of interest. Alternatively, the antibodies can be expressed in cell lines other than hybridoma cell lines. More specifically, sequences encoding particular antibodies can be cloned from cells producing the antibodies and used for transformation of a suitable mammalian host cell.

[0120] Transformation can be by any known method for introducing polynucleotides into a host cell, including, for example, packaging the polynucleotide in a virus or into a viral vector and transducing a host cell with a virus or vector or by transfection procedures known in the art such as those described in U.S. Pat. Nos. 4,399,216, 4,912,040, 4,740,461 and 4,959,455. For example, one or more genes encoding the heavy chain can be expressed in a cell and one or more genes encoding the light chain can be expressed in a second cell. The resulting heavy and light chains can then be fused together to form the antibodies of the present invention using techniques known in the art. Alternatively, genes encoding for parts of the heavy and light chains can be ligated using restriction endonucleases to reconstruct the gene coding for each chain. Such a gene can then be expressed in a cell to produce the antibodies of the present invention.

[0121] The transformation procedure used will depend upon the host to be transformed. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) into liposomes and direct microinjection of the DNA molecule.

[0122] Mammalian cell lines that can be used as hosts for expression are well known in the art and include, but are not limited to, Chinese hamster ovary (CHO) cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells bacterial cells, such as E. coli, yeast cells, such as Saccharomyces cerevisiae, etc.

[0123] Humanized antibodies can also be made using a CDR-grafted approach. Such humanized antibodies are well known in the art. Generally, humanized antibodies are produced by obtaining nucleic acid sequences that encode the variable heavy and variable light sequences of an antibody that binds to the EPO receptor, identifying the complementary determining region or "CDR" in the variable heavy and variable light sequences and grafting the CDR nucleic acid sequences on to human framework nucleic acid sequences. (See, for example, U.S. Pat. Nos. 4,816,567 and 5,225,539).

[0124] The human framework that is selected is one that is suitable for in vivo administration, meaning that it does not exhibit immunogenicity. For example, such a determination can be made by prior experience with in vivo usage of such antibodies and studies of amino acid similarities.

[0125] Methods for cloning nucleic acids are known in the art. These methods involve amplification of the antibody sequences to be cloned using appropriate primers by polymerase chain reaction (PCR). Primers that are suitable for amplifying antibody nucleic acid sequences and specifically murine variable heavy and variable light sequences are known in the art.

[0126] Once the CDRs and FRs of the cloned antibody sequences that are to be humanized are identified, the amino acid sequences encoding the CDRs are identified and the corresponding nucleic acid sequences grafted on to selected human FRs. This can be done using known primers and linkers, the selection of which are known in the art.

[0127] After the CDRs are grafted onto selected human FRs, the resulting "humanized" variable heavy and variable light sequences are expressed to produce a humanized Fv or humanized antibody that binds to the EPO receptor. Typically, the humanized variable heavy and light sequences are expressed as a fusion protein with human constant domain sequences so an intact antibody that binds to the EPO receptor is obtained. However, a humanized Fv antibody can be produced that does not contain the constant sequences. Fusion of the human constant sequence to the humanized variable region is preferred.

[0128] The EPO receptor that is bound by and preferably activated using the antibodies of the present invention is preferably a mammalian EPO receptor, most preferably a human EPO receptor. The present invention also contemplates the use of analogs of the EPO receptor, such as those described in U.S. Pat. No. 5,292,654. Human EPO receptor can be purchased from R & D Systems (Minneapolis, Minn.).

[0129] An example of two (2) antibodies that (1) bind to and activate the EPO receptor; (2) do not interact with a peptide having an amino acid sequence of PGNYSFSYQLEDEPWKLCRLHQAPTARGAV (SEQ ID NO: 1); and (3) exhibit a binding affinity within one hundred fold of the binding affinity of endogeous human EPO to the EPO receptor, are the human antibodies designated Ab12 and Ab198. Abl 2 and Abl 98 are human antibodies that were developed using the XenoMouse.RTM. XenoMax technology described herein (See Example 1).

[0130] In another embodiment, the present invention relates to polynucleotide and polypeptide sequences that encode for the antibodies described herein. Preferably, such polynucleotides encode for both the variable and constant regions of each of the heavy and light chains, although other combinations are also contemplated by the present invention.

[0131] The present invention also contemplates oligonucleotide fragments derived from the disclosed polynucleotides and nucleic acid sequences complementary to these polynucleotides. The polynucleotides can be in the form of RNA or DNA. Polynucleotides in the form of DNA, cDNA, genomic DNA, nucleic acid analogs and synthetic DNA are within the scope of the present invention. The DNA may be double-stranded or single-stranded, and if single stranded, may be the coding (sense) strand or non-coding (anti-sense) strand. The coding sequence that encodes the polypeptide may be identical to the coding sequence provided herein or may be a different coding sequence which coding sequence, as a result of the redundancy or degeneracy of the genetic code, encodes the same polypeptide as the DNA provided herein.

[0132] Preferably, the polynucleotides encode at least one heavy chain variable region and at least one light chain variable region of the present invention. Examples of such polynucleotides are shown in SEQ ID NOS: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 50, 52, 54 and 56 as well as fragments, complements and degenerate codon equivalents thereof. For example, SEQ ID NO: 2 encodes for the heavy chain of Abl 2 (variable region) and SEQ ID NO:4 encodes for the light chain of Abl 2 (variable region). SEQ ID NO:6 encodes for the heavy chain of Abl 98 (variable region) and SEQ ID NO: 8 encodes for the light chain of Abl 98 (variable region).

[0133] The present invention also includes variant polynucleotides containing modifications such as polynucleotide deletions, substitutions or additions, and any polypeptide modification resulting from the variant polynucleotide sequence. A polynucleotide of the present invention may also have a coding sequence that is a naturally occurring variant of the coding sequence provided herein.

[0134] It is contemplated that polynucleotides will be considered to hybridize to the sequences provided herein if there is at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95% identity between the polynucleotide and the sequence.

[0135] The present invention further relates to polypeptides that encode for the antibodies of the present invention as well as fragments, analogs and derivatives of such polypeptides. The polypeptides of the present invention may be recombinant polypeptides, naturally purified polypeptides or synthetic polypeptides. The fragment, derivative or analogs of the polypeptides of the present invention may be one in which one or more of the amino acid residues is substituted with a conserved or non-conserved amino acid residue (preferably a conserved amino acid residue) and such substituted amino acid residue may or may not be one encoded by the genetic code; or it may be one in which one or more of the amino acid residues includes a substituent group; or it may be on in which the polypeptide is fused with another compound, such as a compound to increase the half-life of the polypeptide (for example, polyethylene glycol); or it may be one in which the additional amino acids are fused to the polypeptide, such as a leader or secretory sequence or a sequence that is employed for purification of the polypeptide or a proprotein sequence. Such fragments, derivatives and analogs are within the scope of the present invention.

[0136] A polypeptide of the present invention may have an amino acid sequence that is identical to that of the antibodies described herein or that is different by minor variations due to one or more amino acid substitutions. The variation may be a "conservative change" typically in the range of about 1 to 5 amino acids, wherein the substituted amino acid has similar structural or chemical properties, e.g., replacement of leucine with isoleucine or threonine with serine. In contrast, variations may include nonconservative changes, e.g., replacement of a glycine with a tryptophan. Similar minor variations may also include amino acid deletions or insertions or both. Guidance in determining which and how many amino acid residues may be substituted, inserted, or deleted without changing biological or immunological activity may be found using computer programs well known in the art, for example DNASTAR software (DNASTAR, Inc. Madison, Wis.).

[0137] Preferably, the polypeptides encode at least one heavy chain variable region or at least one light chain variable region of the antibodies of the present invention. More preferably, the polypeptides encode at least one heavy chain variable region and one light chain variable region of the antibodies of the present invention. Examples of such polypeptides are those having the amino acid sequences shown in SEQ ID NOS: 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 46, 47, 49, 51, 53, 55, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, and fragments thereof. Specifically, the heavy chain of Ab12 has the amino acid sequence shown in SEQ ID NO: 3 and the light chain has the amino acid sequence shown in SEQ ID NO:5. The amino acid sequence of the heavy chain of Ab198 is shown in SEQ ID NO:7 and the light chain has the amino acid sequence shown in SEQ ID NO:9.

[0138] The present invention also provides vectors that include the polynucleotides of the present invention, host cells which are genetically engineered with vectors of the present invention and the production of the antibodies of the present invention by recombinant techniques.

[0139] Host cells are genetically engineered (transfected, transduced or transformed) with vectors, such as, cloning vectors or expression vectors. The vector may be in the form of a plasmid, a viral particle, a phage, etc. The engineered host cells can be cultured in conventional nutrient media modified as appropriate for activating promoters, selecting transfected cells, etc. The culture conditions, such as temperature, pH and the like, are those previously used with the host cell selected for expression, and will be apparent to those of skilled in the art.

[0140] The polynucleotides of the present invention can be employed to produce the polypeptides and hence the antibodies of the present invention. The polynucleotide sequences of the present invention can be included in any one of a variety of expression vehicles, in particular, vectors or plasmids for expressing a polypeptide. Such vectors include chromosomal, nonchromosomal and synthetic DNA sequences, derivatives of SV40, bacterial plasmids, phage DNA, yeast plasmids, vectors derived from combinations of plasmids and phage DNA, viral DNA such as vaccinia, adenovirus, fowl pox virus and pseudorabies. However, any other plasmid or vector may be used so long as it is replicable and viable in the host.

[0141] The appropriate DNA sequence may be inserted into the vector by a variety of procedures. In general, the DNA sequence is inserted into appropriate restriction endonuclease sites by procedures known in the art. The polynucleotide sequence in the expression vector is operatively linked to an appropriate expression control sequence (i.e. promoter) to direct mRNA synthesis. Examples of such promoters include, but are not limited to, the LTR or the SV40 promoter, the E. coli lac or trp, the phage lambda P.sub.L promoter and other promoters known to control expression of genes in prokaryotic or eukaryotic cells or their viruses. The expression vector also contains a ribosome binding site for translation initiation and a transcription terminator. The vector may also include appropriate sequences for amplifying expression. For example, the vector can contain enhancers, which are transcription-stimulating DNA sequences of viral origin, such as those derived form simian virus such as SV40, polyoma virus, bovine papilloma virus or Moloney sarcoma virus, or genomic, origin. The vector preferably also contains an origin of replication. The vector can be constructed to contain an exogenous origin of replication or, such an origin of replication can be derived from SV40 or another viral source, or by the host cell chromosomal replication mechanism.

[0142] In addition, the vectors preferably contain a marker gene for selection of transfected host cells such as dihydrofolate reductase or antibiotics, such as G-418 (geneticin, a neomycin-derivative) or hygromycin, or genes which complement a genetic lesion of the host cells such as the absence of thymidine kinase, hypoxanthine phosphoribosyl transferase, dihydrofolate reductase, etc.

[0143] Suitable vectors for use in the present invention are known in the art. Any plasmid or vector can be used in the present invention as long as it is replicable and is viable in the host. Examples of vectors that can be used include those that are suitable for mammalian hosts and based on viral replication systems, such as simian virus 40 (SV40), Rous sarcoma virus (RSV), adenovirus 2, bovine papilloma virus (BPV), papovavirus BK mutant (BKV), or mouse and human cytomegalovirus (CVM).

[0144] In a further embodiment, the present invention relates to host cells containing the above-described constructs. The host cell can be a higher eukaryotic cell, such as a mammalian cell, or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. Preferably, the host cells provide a suitable environment for the production of active antibodies, since the biosynthesis of functional tetrameric antibody molecules requires correct nascent polypeptide chain folding, glycosylation, and assembly. Example of suitable host cells, include mammalian cells, such as COS-7 cells, Bowes melanoma cells, Chinese hamster ovary (CHO) cells, embryonic lung cells L-132, and mammalian cells of lymphoid origin, such as myeloma or lymphoma cells. The host cells can be transfected with a vector containing a polynucleotide sequence encoding the H-chain alone, with a second vector encoding the light chain alone (such as by using two different vectors as discussed previously). Preferably, the host cells are transfected with two different vectors.

[0145] Introduction of the vectors into the host cell can be effected by calcium phosphate transfection, DEAE-Dextran mediated transfection or electroporation (L. David et al., Basic Methods in Molecular Biology 2.sup.nd Edition, Appleton and Lang, Paramount Publishing, East Norwalk, Conn. (1994)).

[0146] In order to obtain the antibodies of the present invention, one or more polynucleotide sequences that encode for the light and heavy chain variable regions and light and heavy chain constant regions of the antibodies of the present invention should be incorporated into a vector. Polynucleotide sequences encoding the light and heavy chains of the antibodies of the present invention can be incorporated into one or multiple vectors and then incorporated into the host cells.

[0147] Cell lines expressing Ab12 and Ab467 antibodies were deposited with the American Type Culture Collection (ATCC), 10801 University Boulevard, Manassas, Va. 20110, under the terms of the Budapest Treaty, on Sep. 30, 2003 and were accorded accession numbers PTA-5554 and PTA-5555. These deposits are provided for the convenience of those skilled in the art and are neither an admission that such deposits are required to practice the invention nor that equivalent embodiments are not within the skill of the art in view of the present disclosure. The public availability of these deposits is not a grant of a license to make, use or sell the deposited materials under this or any other patents. The nucleic acid sequences of the deposited materials are incorporated in the present disclosure by reference and are controlling if in conflict with any sequence described herein.

[0148] The antibodies of the present invention have a number of uses. In general, the antibodies may be used to treat any condition treatable by erythropoietin or a biologically active variant or analog thereof. For example, antibodies of the invention are useful for treating disorders characterized by low red blood cell levels and/or decreased hemoglobin levels (e.g. anemia). In addition, the antibodies of the invention may be used for treating disorders characterized by decreased or subnormal levels of oxygen in the blood or tissue, such as, for example, hypoxemia or chronic tissue hypoxia and/or diseases characterized by inadequate blood circulation or reduced blood flow. Antibodies of the invention also may be useful in promoting wound healing or for protecting against neural cell and/or tissue damage, resulting from brain/spinal cord injury, stroke and the like. Non-limiting examples of conditions that may be treatable by the antibodies of the invention include anemia, such as chemotherapy-induced anemia, cancer associated anemia, anemia of chronic disease, HIV-associated anemia, bone marrow transplant-associated anemia and the like, heart failure, ischemic heart disease and renal failure. As such, the invention includes methods of treating any of the aforementioned diseases or conditions comprising the step of administering to a mammal a therapeutically effective amount of said antibody. Preferably, the mammal is a human.

[0149] The antibodies of the present invention also can be used to identify and diagnose mammals that have a dysfunctional EPO receptor. Mammals that have a dysfunctional EPO receptor are characterized by disorders such as anemia.

[0150] Preferably, the mammal being identified and diagnosed is a human. Additionally, the antibodies of the present invention can be used in the treatment of anemia in mammals suffering from red blood cell aplasia. Red blood cell aplasia may result from the formation of neutralizing anti-erythropoietin antibodies in patients during treatment with recombinant erythropoietin (Casadevall, N. et al., n. Eng. J. Med. 346: 469 (2002)). The method involves the step of administering to a mammal suffering from said aplasia and in need of treatment a therapeutically effective amount of the antibodies of the present invention.

[0151] In another embodiment of the invention, the EPO receptor antibodies and antibody fragments of the invention also can be used to detect EPO receptor (e.g., in a biological sample, such as tissue specimens, intact cells, or extracts thereof), using a conventional immunoassay, such as an enzyme linked immunosorbent assay (ELISA), a radioimmunoassay (RIA) or tissue immunohistochemistry. The invention provides a method for detecting EPO receptor in a biological sample comprising contacting a biological sample with an antibody or antibody fragment of the invention and detecting either the antibody (or antibody portion), to thereby detect EPO receptor in the biological sample. The antibody or antibody fragment is directly or indirectly labeled with a detectable substance to facilitate detection of the bound or unbound antibody or antibody fragment. A variety of immunoassay formats may be practiced (such as competitive assays, direct or indirect sandwich immunoassays and the like) and are well known to those of ordinary skill in the art.

[0152] Suitable detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, B-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine, dansyl chloride or phycoerythrin; and an example of a luminescent material includes luminol; and examples of suitable radioactive material include .sup.125I, .sup.131I, .sup.35S, or .sup.3H.

[0153] In yet another embodiment, the present invention relates to a pharmaceutical composition containing a therapeutically effective amount of the antibody of the present invention along with a pharmaceutically acceptable carrier or excipient. As used herein, "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, coating, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible. Examples of pharmaceutically acceptable carriers or excipients include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol and the like as well as combinations thereof. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Pharmaceutically acceptable substances such as wetting or minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the of the antibody or antibody portion also may be included. Optionally, disintegrating agents can be included, such as cross-linked polyvinyl pyrrolidone, agar, alginic acid or a salt thereof, such as sodium alginate and the like. In addition to the excipients, the pharmaceutical composition can include one or more of the following, carrier proteins such as serum albumin, buffers, binding agents, sweeteners and other flavoring agents; coloring agents and polyethylene glycol.

[0154] The compositions of this invention may be in a variety of forms. They include, for example, liquid, semi-solid and solid dosage forms, such as liquid solutions (e.g. injectable and infusible solutions), dispersions or suspensions, tablets, pills, powders, liposomes and suppositories. The preferred form depends on the intended mode of administration and therapeutic application. Typical preferred compositions are in the form of injectable or infusible solutions, such as compositions similar to those used for passive immunization of humans with other antibodies. The preferred mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In a preferred embodiment, the antibody is administered by intravenous infusion or injection. In another preferred embodiment, the antibody or antibody fragment is administered by intramuscular or subcutaneous injection.

[0155] Other suitable routes of administration for the pharmaceutical composition include, but are not limited to, rectal, transdermal, vaginal, transmucosal or intestinal administration.

[0156] Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, dispersion, liposome, or other ordered structure suitable to high drug concentration. Sterile injectable solutions can be prepared by incorporating the active compound (i.e. antibody or antibody fragment) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. The proper fluidity of a solution can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Prolonged absorption of injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, monostearate salts and gelatin.

[0157] The antibodies and antibody fragments of the invention can be administered by a variety of methods known in the art, although for many therapeutic applications, the preferred route/mode of administration is intravenous injection or infusion. As will be appreciated by the skilled artisan, the route and/or mode of administration will vary depending upon the desired results. In certain embodiments, the active compound may be prepared with a carrier that will protect the compound against rapid release, such as a controlled release formulation, including implants, transdermal patches, and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Many methods for the preparation of such formulations are patented or generally known to those skilled in the art. See, e.g. Sustained and Controlled Release Drug Delivery Systems, J. R. Robinson, ed., Marcel Dekker, Inc., New York, 1978).

[0158] In certain embodiments, an antibody or antibody portion of the invention may be orally administered, for example, with an inert diluent or an assimilable edible carrier. The compound (and other ingredients if desired) may also be enclosed in a hard or soft shell gelatin capsule, compressed into tablets, buccal tablets, troches, capsules, elixiers, suspensions, syrups, wafers, and the like. To administer an antibody or antibody fragment of the invention by other than parenteral administration, it may be necessary to coat the compound with, or co-administer the compound with, a material to prevent its inactivation.

[0159] Supplementary active compounds also can be incorporated into the compositions. In certain embodiments, an antibody or antibody fragment of the invention is coformulated with and/or coadministered with one or more additional therapeutic agents. For example, an EPO receptor antibody or antibody fragment of the invention may be coformulated and/or coadministerd with one or more additional antibodies that bind other targets (e.g., antibodies that bind other cytokines or that bind cell surface molecules) or one or more cytokines. Furthermore, one or more antibodies of the invention may be used in combination with two or more of the foregoing therapeutic agents. Such combination therapies may advantageously utilize lower dosages of the administered therapeutic agents, thus avoiding possible toxicities or complications associated with the various monotherapies.

[0160] As used herein, the term "therapeutically effective amount" means an amount of antibody or antibody fragment that produces the effects for which it is administered. The exact dose will be ascertainable by one skilled in the art. As known in the art, adjustments based on age, body weight, sex, diet, time of administration, drug interaction and severity of condition may be necessary and will be ascertainable with routine experimentation by those skilled in the art. A therapeutically effective amount is also one in which the therapeutically beneficial effects outweigh any toxic or detrimental effects of the antibody or antibody fragment. A "prophylactically effective amount" refers to an amount effective, at dosages and for periods of time necessary to achieve the desired prophylactic result. Typically, since a prophylactic dose is used in subjects prior to or at an earlier stage of disease, the prophylactically effective amount will be less than the therapeutically effective amount.

[0161] Dosage regimens may be adjusted to provide the optimum desired response (e.g., a therapeutic or prophylactic response). For example, a single bolus may be administered, several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be tested; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active compound and the particular therapeutic or prophylactic effect to be achieved and (b) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals.

[0162] An exemplary, non-limiting range for a therapeutically or prophylactically effective amount of an antibody or antibody portion of the invention is 0.1-20 mg/kg, more preferably 1-10 mg/kg. It is to be noted that dosage values may vary with the type and severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions, and that dosage ranges set forth herein are exemplary only and are not intended to limit the scope or practice of the claimed composition.

[0163] By way of example, and not of limitation, examples of the present invention shall now be given.

Example 1

Generation of Human Erythropoietin Receptor Antibodies

[0164] Antigen Preparation. The antigen used for immunization of XenoMouse.RTM. animals was coupled to a universal T-cell epitope (TCE) (J. Immunol., 148(5):1499 (1992)) using two different methods. A mixture containing an equal amount of each was used as the immunogen.

[0165] 1) 2.3 mg of Dithiothreitol (DTT), and 200 mcg of cysteine coupled TCE (J. Immunol., 148(5):1499 (1992)) are mixed at room temperature for 30 minutes. DTT is removed by centrifugation through a Sephadex G10 (Pharmacia, Upsala, Sweden) chromatography column. The reduced cysteine coupled TCE is added to 200 mcg soluble extracellular domain of human EpoR (R&D Systems, Minneapolis Minn.) re-suspended in Phosphate Buffered Saline (PBS) (8.1 mM Na.sub.2HPO.sub.4, 1.6 mM NaH.sub.2PO.sub.4, 136 mM NaCl, 2.6 mM KCl, pH 7.4) and 33 mcg of Sulfosuccinimidyl 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (sulfo SMCC), and mixed 4.degree. C. over night. Un-reacted EpoR was removed by centrifugation through a 10KDa cut off Centricon column (Millipore, Bedford, Mass.).

[0166] 2) The soluble extracellular domain (200 mcg) of human EpoR (R&D Systems, Minneapolis, Minn.) was re-suspended in PBS and mixed with 4 mcg of TCE-BPA (p-Benzoyl Phenylalanine) and incubated under UV light (362 nM) at room temperature for 45 minutes. The un-reacted EpoR was removed by centrifugation through a 10KDa cut off Centricon column (Millipore, Bedford, Mass.).

[0167] Immunization of animals. Monoclonal antibodies of the invention, including Ab12 and Ab198 (also referred to herein as AB-ABT2-XG2-012 and AB-ABT2-XG2-198, respectively) were developed by immunizing XenoMouse.RTM. mice (XenoMouse.RTM. XG2, Abgenix, Inc., Fremont, Calif. and Vancouver, BC) with soluble EpoR coupled to a TCE as described above. The initial immunization was with 20 mcg of antigen and mixed 1:1 v/v with Complete Freund's Adjuvant (CFA) (Sigma, St Louis, Mo.) per mouse. The subsequent immunizations were with 20 mcg of antigen mixed 1:1 v/v with incomplete Freund's (IFA). In particular, each animal was immunized at the base of tail and by intraperitoneal injection on days 0, 14, 28 and 42.

[0168] Biotinylation of EpoR. 300 mcg of EpoR (Abbott CHO cell derived ref. # RB69084:4) was re-suspended in 990 mcL of PBS pH 8.6 and added to 100 mcg of biotin-NHS (Pierce, Rockford, 1H) dissolved in DMSO (Dimethyl Sulfoxide) incubated for forty minutes at room temperature (RT). Free biotin and buffer was removed by centrifugation through a 5 kDa Centricon column with several washes with PBS pH 7.4 and re-suspended in an appropriate volume to a final concentration was 600 mcg/mL.

[0169] Selection of animals for harvest. Anti-EpoR antibody titers were determined by ELISA. 0.7 mcg/ml biotin EpoR (described above) was coated onto streptavadin plates (Sigma, St Louis, Mo.) at room temperature for 1 hour. The solution containing unbound biotin EpoR was removed and all plates were washed five times with dH.sub.2O. XenoMouse.RTM. sera from the EpoR immunized animals, or naive XenMouse.RTM. animals, were titrated in 2% milk/PBS at a 1:2 dilution in duplicate from a 1:100 initial dilution. The last well was left blank, and plates were washed five times with dH.sub.2O. A goat anti-human IgG Fc-specific horseradish peroxidase (HRP) (Pierce, Rockford, Ill.) conjugated antibody was added at a final concentration of 1 mcg/mL for 1 hour at room temperature. The plates were washed five times with dH.sub.2O. The plates were developed with the addition of TMB chromogenic substrate (KPL, Gaithersburg, Md.) for 30 minutes and the ELISA was stopped by the addition of 1 M phosphoric acid. The specific titers obtained from XenoMouse.RTM. animals were determined from the optical density at 450 nm and are shown in Table 1. The titer represents the reciprocal dilution of the serum and therefore the higher the number the greater the humoral immune response to EpoR.

TABLE-US-00001 TABLE 1 Mouse I.D. Titer 11 1600 12 12800 13 51200 14 102400 15 102400 16 0 17 102400 18 3200 19 102400 20 2560

XenoMouse.RTM. animal 14 was selected for harvest based on the serology data in Table 1.

[0170] Culture and selection of B cells. B cells from the harvested animals were cultured and those secreting EpoR-specific antibodies were isolated essentially as described in Babcook et al., Proc. Natl. Acad. Sci. USA, 93:7843-7848 (1996). ELISA, performed as described above for sera titers, was used to identify EpoR-specific wells. Fifty plates cultured at 500 cells/well were screened on biotin EpoR to identify the antigen-specific wells. The data as shown in Table 2 demonstrated the presence of 701 wells with ODs significantly over background (0.05).

TABLE-US-00002 TABLE 2 Optical Density Number of Positives 0.1 701 0.2 273 0.3 163 0.4 130 0.5 102 0.6 91 0.7 76 0.8 70 0.9 67 1.0 65 2.0 25 3.0 7

[0171] These data indicated a very low frequency of hits and indicated that the wells were monoclonal for antigen-specificity. These 701 positive wells were re-screened on biotin EpoR and 137 wells (shown in bold in Table 3 below) were found to repeat as real antigen-specific wells with ODs significantly over background (0.05).

TABLE-US-00003 TABLE 3 Optical Density Number of Positives 0.1 207 0.15 137 0.2 110 0.3 94 0.4 85 0.5 79 0.6 71 0.7 63 0.8 57 0.9 53 1.0 50 2.0 32 3.0 13

[0172] Agonist activity assay. Proliferation of an Epo responsive cell line was used as the basis for the agonist screen. These 137 wells were then screened for agonist activity using the human erythroleukemia cell line UT-7/Epo (Abbott ref #.RB29454-174). 12.5 mcL of supernatant were added to 1.times.105 cells per well in RPMI 1640 (10% FCS) to a final volume of 50 mcL in a half-area 96 well plate. The well size is half the area of a typical 96 well plate. Proliferation was identified visually and compared to cells in media containing a titration of human Epo or no Epo as a base line control. Eleven wells with proliferation activity were identified.

[0173] EpoR-specific Hemolytic Plaque Assay. A number of specialized reagents are needed to conduct the assay. These reagents were prepared as follows.

[0174] Biotinylation of Sheep red blood cells (SRBC): SRBC are stored in RPMI media as a 25% stock. A 250 ul SRBC packed-cell pellet was obtained by aliquoting 1.0 ml of the stock into a 15-ml falcon tube, spinning down the cells and removing the supernatant. The cell pellet was then re-suspended in 4.75 ml PBS at pH 8.6 in a 50 ml tube. In a separate 50 ml tube, 2.5 mg of Sulfo-NHS biotin was added to 45 ml of PBS at pH 8.6. Once the biotin had completely dissolved, 5 ml of SRBCs were added and the tube rotated at RT for 1 hour. The SRBCs were centrifuged at 3000 g for 5 min, the supernatant drawn off and 25 mls PBS at pH 7.4 as a wash. The wash cycle was repeated 3 times, then 4.75 ml immune cell media (RPMI 1640 with 10% FCS) was added to the 250 ul biotinylated-SRBC (B-SRBC) pellet to gently re-suspend the B-SRBC (5% B-SRBC stock). Stock was stored at 4.degree. C. until needed.

[0175] Streptavidin (SA) coating of B-SRBC: One ml of the 5% B-SRBC stock was transferred into to a fresh eppendorf tube. The B-SRBC cells were pelleted with a pulse spin at 8000 rpm (6800 rcf) in a microfuge, the supernatant drawn off, the pellet re-suspended in 1.0 ml PBS at pH 7.4, and the centrifugation repeated. The wash cycle was repeated 2 times, then the B-SRBC pellet was resuspended in 1.0 ml of PBS at pH 7.4 to give a final concentration of 5% (v/v). 10 ul of a 10 mg/ml streptavidin (CalBiochem, San Diego, Calif.) stock solution was added and the tube mixed and rotated at RT for 20 min. The washing steps were repeated and the SA-SRBC were re-suspended in 1 ml PBS pH 7.4 (5% (v/v)).

[0176] EpoR coating of SA-SRBC: The SA-SRBC were coated with biotinylated EpoR at 10 ug/ml, the mixed and rotated at RT for 20 min. The SRBC were washed twice with 1.0 ml of PBS at pH 7.4 as above. The EpoR-coated SRBC were re-suspended in RPMI (+10% FCS) to a final concentration of 5% (v/v).

[0177] Determination of the quality of EpoR-SRBC by immunofluorescence (IF): 10 ul of 5% SA-SRBC and 10 ul of 5% PTH-coated SRBC were each added to separate fresh 1.5 ml eppendorf tube containing 40 ul of PBS. The murine anti-EpoR antibody (R&D Systems Cat. # MAB307) was added to each sample of SRBCs at 20 ug/ml. The tubes were rotated at RT for 25 min. and the cells were then washed three times with 100 ul of PBS. The cells were re-suspended in 50 ul of PBS and incubated with 40 mcg/mL Gt-anti mouse IgG Fc antibody conjugated to Alexa488 (Molecular Probes, Eugene, Oreg.). The tubes were rotated at RT for 25 min, and then washed with 100 ul PBS and the cells re-suspended in 10 ul PBS. 10 ul of the stained cells were spotted onto a clean glass microscope slide, covered with a glass coverslip, observed under fluorescent light, and scored on an arbitrary scale of 0-4.

[0178] Preparation of Plasma Cells: the Contents of a Single Microculture Well identified by the previous assays as containing a B cell clone secreting the immunoglobulin of interest were harvested. Using a 100-1000 ul pipettman, the contents of the well were recovered by adding 37C RPMI (+10% FCS). The cells were re-suspended by pipetting and then transferred to a fresh 1.5 ml eppendorf tube (final vol. approx 500-700 ul). The cells were centrifuged in a microfuge at 1500 rpm (240 rct) for 2 minutes at room temperature, then the tube rotated 180 degrees and spun again for 2 minutes at 1500 rpm. The freeze media was drawn off and the immune cells resuspended in 100 ul RPMI (10% FCS), then centrifuged. This washing with RPMI (10% FCS) was repeated and the cells re-suspended in 60 ul RPMI (FCS) and stored on ice until ready to use.

[0179] Plaque assay: Glass slides (2.times.3 inch) were prepared in advance with silicone edges and allowed to cure overnight at RT. Before use the slides were treated with approx. 5 ul of SigmaCoat (Sigma, Oakville, ON) wiped evenly over glass surface, allowed to dry and then wiped vigorously. To a 60 ul sample of cells was added 60 ul each of EpoR-coated SRBC (5% v/v stock), 4.times. guinea pig complement (Sigma, Oakville, ON) stock prepared in RPMI with 10% FCS, and 4.times. enhancing sera stock (1:900 in RPMI with 10% FCS). The mixture (3-5 ul) was spotted onto the prepared slides and the spots covered with undiluted paraffin oil. The slides were incubated at 37.degree. C. for a minimum of 45 minutes.

[0180] Plague assay results: The coating was determined qualitatively by immunofluorescent microscopy to be very high (4/4) using MAB307 to detect coating compared to a secondary detection reagent alone (0/4). There was no signal detected using the MAB307 antibody on red blood cells that were only coated with streptavidin (0/4). These red blood cells were then used to identify antigen-specific plasma cells from the fourteen wells identified in Table 4. After micromanipulation to rescue the antigen-specific plasma cells, the genes encoding the variable region genes were rescued by RT-PCR on a single plasma cell.

TABLE-US-00004 TABLE 4 Plate ID Single Cell numbers 11G10 ABT2-SCX-251-260 21D1 ABT2-SCX-54 25C3 ABT2-SCX-134-144 29G8 ABT2-SCX-1-11 33G8 ABT2-SCX-12-18 37A11 ABT2-SCX-19-44 43H12 ABT2-SCX-185-201, 233-239 16F7 ABT2-SCX-267-278 24C3 ABT2-SCX-55-77 24F8 ABT2-SCX-82-102 34D4 ABT2-SCX-145-168

[0181] Expression. After isolation of the single plasma cells, mRNA was extracted and reverse transcriptase PCR was conducted to generate cDNA. The cDNA encoding the variable heavy and light chains was specifically amplified using polymerase chain reaction. The variable heavy chain region was cloned into an IgG2 expression vector. This vector was generated by cloning the constant domain of human IgG2 into the multiple cloning site of pcDNA3.1+/Hygro (Invitrogen, Burlington, ON). The variable light chain region was cloned into an IgK expression vector. This vector was generated by cloning the constant domain of human IgK into the multiple cloning site of pcDNA3.1+Neo (Invitrogen, Burlington, ON). The appropriate pairs of heavy chain and the light chain expression vectors were then co-lipofected into a 60 mm dish of 70% confluent human embryonal kidney 293 cells and the transfected cells were left to secrete a recombinant antibody for 24 hours. The supernatant (3 mL) was harvested from the HEK 293 cells and the secretion of an intact antibody (AB-ABT2-XG2-012 and AB-ABT2-XG2-198) was demonstrated with a sandwich ELISA to specifically detect human IgG (Table 5, fourth column). The specificity of AB-ABT2-XG2-012 and AB-ABT2-XG2-198 was assessed through binding of the recombinant antibody to biotinylated EpoR using ELISA (Table 5, fifth column).

TABLE-US-00005 TABLE 5 Well ID Single cell number Secretion Binding 11G10 ABT2-SCX-254 1:4 1:8 21D1 ABT2-SCX-054 >1:64 >1:64 25C3 ABT2-SCX-135 1:4 1:4 29G8 ABT2-SCX-003 >1:64 >1:64 33G8 ABT2-SCX-012 >1:64 >1:64 37A11 ABT2-SCX-022 >1:64 >1:64 43H12 ABT2-SCX-198 >1:64 >1:64 16F7 ABT2-SCX-267 >1:64 >1:64 24C3 ABT2-SCX-060 >1:64 >1:64 24F8 ABT2-SCX-102 >1:64 >1:64 34D4 ABT2-SCX-145 >1:64 >1:64

[0182] The ELISA for antigen specific antibody secretion was performed as follows. Control plates were coated with 2 mg/mL Goat anti-human IgG H+L O/N. For the binding plates, biotin-EpoR (0.7 mcg/mL) was coated onto streptavadin 96 well plates (Sigma, St Louis, Mo.) for one hour at room temperature. The plates were washed five times with dH.sub.2O. Recombinant antibodies were titrated 1:2 for 7 wells from the undiluted minilipofection supernatant. The plates were washed five times with dH.sub.2O. A goat anti-human IgG Fc-specific HRP-conjugated antibody was added at a final concentration of 1 ug/mL for 1 hour at RT for the secretion and the binding ELISA. The plates were washed five times with dH.sub.2O. The plates were developed with the addition of TMB chromogenic substrate (KPL, Gaithersburg, Md.) for 30 minutes and the ELISA was stopped by the addition of 1 M phosphoric acid. Each ELISA plate was analyzed to determine the optical density of each well at 450 nm.

[0183] Purification of AB-ABT2-XG2-012 and AB-ABT2-XG2-198. For larger scale production, the heavy and light chain expression vectors (2.5 ug of each chain/dish) were lipofected into ten 100 mm dishes that were 70% confluent with HEK 293 cells. The transfected cells were incubated at 37.degree. C. for 4 days, the supernatant (6 mL) was harvested and replaced with 6 mL of fresh media. At day 7, the supernatant was removed and pooled with the initial harvest (120 mL total from 10 plates). The ABT2-XG2-012 and ABT2-XG2-198 antibody were purified from the supernatant using a Protein-A Sepharose (Amersham Biosciences, Piscataway, N.J.) affinity chromatography (1 mL). The antibody was eluted from the Protein-A column with 500 mcL of 0.1 M Glycine pH 2.5. The eluate was dialysed in PBS pH 7.4 and filter sterilized. The antibody was analyzed by non-reducing SDS-PAGE to assess purity and yield.

[0184] Agonist activity of recombinant antibodies. The ability of these recombinant antibodies to stimulate the proliferation of Epo responsive cells was examined using the UT-7/Epo cells with proliferation quantitated by MTS reagent (Promega, Madison, Wis.) measured at 490 nm as described in the Agonist Activity Assay above. ABT2-SCX-012 and ABT2-SCX-198 induced proliferation in comparison to cells in media without antibody and are shown below (FIGS. 14 and 15 respectively).

[0185] Effect of anti-Human Fc. It is possible that the agonist activity of ABT2-SCX-012 and ABT2-SCX-198 are due to self-aggregation. In order to address this issue we induced aggregation by the addition of an anti-human Fc secondary antibody and the effect on the agonist activity of ABT2-SCX-012 and ABT2-SCX-198 was determined using the UT-7/Epo cells. As shown below the addition of a secondary antibody had no effect on the activity of ABT2-SCX-198 (FIG. 16) and inhibited the activity of ABT2-SCX-012 (FIG. 4 17).

[0186] Since the addition of secondary Ab inhibited the activity of ABT2-SCX-012 we concluded that aggregation of this antibody interferes with it's activity and thus it is unlikely that ABT2-SCX-012 has agonist activity due to aggregation. However, the results of ABT2-SCX-198 are more difficult to interpret. The lack of an effect could suggest that ABT2-SCX-198 is fully aggregated and thus the addition of secondary Ab has no further effects on its activity. Alternatively, the lack of effect suggests the activity of ABT2-SCX-198 is not perturbed by the conformational restrictions applied by a secondary antibody.

[0187] Sequence analysis of ABT2-SCX-012 and ABT2-SCX-198 The variable heavy chains and the variable light chains for antibodies ABT2-SCX-012 and ABT2-SCX-198 were sequenced to determine their DNA sequences. The complete sequence information for the anti-EpoR antibodies shown in FIGS. 1, 2, and 18-30 with nucleotide and amino acid sequences for each variable region of the heavy chain gamma and kappa light chains. FIGS. 1 and 2 provide full-length sequences, including the constant regions.

[0188] The variable heavy sequences were analyzed to determine the VH family, the D-region sequence and the J-region sequence. The sequences were then translated to determine the primary amino acid sequence (FIG. 29) and compared to the germline VH, D and J-region sequences to assess somatic hypermutations. The primary amino acid sequences of all the anti-EpoR antibody gamma chains are shown in FIG. 16. The germline sequences are shown above and the mutations are indicated with the new amino acid sequence. Unaltered amino acids are indicated with a dash (-). The light chain was analyzed similarly to determine the V and the J-regions and to identify any somatic mutations from germline kappa sequences (FIG. 30). The heavy chain of ABT2-SCX-012 was shown to utilize the VH 4-59 (DP-71), DlR4rc and the JH4a gene segments, while the light chain was shown to use the VkI (A30) and the Jk1 gene segments. The heavy chain of ABT2-SCX-198 was shown to utilize the VH 3-30 (V3-30), D4-23 and the JH6b gene segments, while the light chain was shown to use the VkI (L5) and the Jk3 gene segments.

Example 2

Competition of Ab12 with .sup.125I-Labeled EPO for Binding CHO Cells Expressing Recombinant EPO Receptor

[0189] CHO cells expressing the full length recombinant human EPO receptor were plated at 5.times.10.sup.5 cells/well in 24 well plates 72 hours prior to the assay. On the day of the assay, 95 ul of Abl 2, Abl 98, or EPO at indicated concentrations (shown in FIG. 5) diluted in RPMI 1640, 0.5% BSA, 1 mM Na N.sub.3 and 5 ul (6 ng) of .sup.125I-EPO (Amersham Cat. #IM178, Arlington Heights, Ill. 486 ci/mM) were added to the wells. After incubating at 37.degree. C. for 1.5 hours, the wells were washed three times with cold HBSS and harvested using 0.5 ml 0.1N NaOH. Samples were counted in a Micromedic ME Plus gamma counter. The results are shown in FIG. 5. Specifically, the results show that Abs 12 and 198 competed with EPO for binding to the erythropoietin receptor.

Example 3

Biacore Studies

[0190] The studies described below were performed on a Biacore 2000 utilizing the Biacontrol software version 3.1. (Biacore, Uppsala, Sweden). Binding analyses were performed with antibody immobilized directly to the chip surface and followed by injection of varying receptor concentrations.

Immobilization of Antibody

[0191] Immobilizations of antibody were performed using the default immobilization program in the Biacore software package. Antibodies were diluted to 10 ug/mL in the supplied acetate buffers to prescreen for the appropriate pH at which to conduct the immobilizations. For immobilizations, antibodies were diluted into the appropriate acetate buffer (10 mM acetate pH 4.0) and coupled directly to the chip surface using standard EDC chemistry at three different protein levels (500, 1000, and 1500 RU). The fourth flow cell was mock coupled with EDC to cap the carboxyl groups and provide a background surface as a negative control.

Binding Studies

[0192] Binding studies were performed by successive injections of varying concentrations of soluble human EPO receptor over the chip surface (500 RU immobilized protein). Binding analyses were performed in the supplied HBS-EP buffer [HBS buffer-10 mM HEPES pH=7.4, 150 mM NaCl, 3 mM EDTA, 0.005% Polysorbate 20 (v/v), Biacore] using receptor diluted to the desired concentrations (10-200 nM) using the running buffer (HBS-EP). Experiments were performed at a flow rate of 30 uL/min. The receptor was injected over a period of 3 minutes followed by a 15 minute dissociation period. Simultaneous injections over the flow cell created as a negative control were also performed. All injections were performed in triplicate.

Model Fitting

[0193] Data were fit to the models available in the BiaEvaluation 3.0.2 software package (Biacore). The data points from the experimental injections were corrected by subtraction of data points from simultaneous over the negative control surface. The corrected data were used to fit to the 1:1 (Langmuir) binding model as well as the bivalent analyte model available in the BiaEvaluation software package. Dissociation constants were calculated directly from fitting to the Langmuir binding model. For the bivalent analyte model, the dissociation constants were calculated indirectly using the calculated values for the kinetic dissociation and kinetic association constants, kd and ka.

TABLE-US-00006 TABLE 6 Antibody kD Ab 12 17.5 nM Ab 198 13.9 nM

Example 4

EPO Dependent Human Cell Proliferation Assay

[0194] Stock cultures of the human erythroleukemic cell line, F36E cells were maintained in RPMI 1640 media with 10% fetal bovine serum and 1 unit per in L of recombinant human erythropoietin. Prior to assays, cells were cultured overnight at a density of 4.0 to 5.0.times.10.sup.5 cells per mL in growth medium without EPO. Cells were recovered, washed and resuspended at a density of 1.0.times.10.sup.6 cells per mL in assay medium (RPMI 1640+10% FBS) and 50 uL of cells added to wells of a 96 well microtiter plate. 50 uL of each of Ab12, Ab 390, Ab 412, Ab 467, Ab 484, Ab 430/432 and Ab198 or EPO standards (recombinant human EPO (rHuEPO)) in assay medium were added to wells and the plates were incubated in a humidified incubator at 37.degree. C. with a 5% CO.sub.2 atmosphere. After 72 hours, 20 .mu.L of Promega Cell Titer 96 Aqueous.RTM. reagent (as prepared per manufacturer's instructions, Madison, Wisconsin) was added to all wells. Plates were incubated at 37.degree. C. with a 5% CO.sub.2 atmosphere for 4 hours and the optical density at 490 nm was determined using a microplate reader (Wallac Victor 1420 Multilabel Counter, Wallac Company, Boston, Mass.). The results are shown in FIG. 6. All Abs stimulated proliferation of the F36E cell line. Maximal proliferative activity was similar to that observed with the EPO control and shown by a bell shaped curve as concentration increased. The results in FIG. 7 demonstrate that Ab12, after storage at 4.degree. C. for up to 20 days, is active in inducing the proliferation of F36E cells. Proliferative activity was similar to that observed with the EPO control with the maximal response differing about ten-fold on a molar equivalent basis

Example 5

Human CD36+ CFUe Assay

[0195] Frozen human CD36+ erythroid progenitor cells obtained from Poietics (Biowhittaker (Walkersville, Md.)) were thawed and 10.sup.4 cells/ml in IMDM-2% FBS. Cells (0.3 ml) were added to 0.3 ml tubes containing 2.4 ml Methocult (StemCell Technologies, Vancouver. Canada) Cat. #04230), 0.3 ml stem cell growth factor (Sigma, St. Louis, Mo. Cat. #S7901, 100 ug/ml), and 0.3 ml EPO(R&D Systems), Ab 12, or IMDM-2% FBS. After mixing, 11.1 ml of the Methocult suspension was added to a 35 mm non tissue culture treated sterile petri dish and incubated at 37.degree. C., 5% CO.sub.2 for 2 weeks. Colonies were identified microscopically. The results are shown in FIG. 8. Specifically, Ab12 induced the formation of CFU-E colonies from human CD 36+ progenitor cells. The colonies, identified microscopically, were red in color. The size and number of the colonies is reduced compared to those observed with the EPO control probably due to a reduced proliferative signal.

Example 6

Demonstration of Erythopoietic Activity in Liquid Cultures

[0196] CD34+ cells were enriched from human peripheral blood using a Direct CD34+ Progenitor Cell Isolation Kit (Miltenyi, Auburn, Calif.). Recovered cells were washed twice with alpha-medium and re-suspended in suspension culture media (alpha-media supplemented with 30% FCS, 1% deionized BSA, 10.sup.-5M .beta.-mercaptoethanol, 10.sup.-6 M dexamethasone, 0.3 mg/mL human hollo-transferrin and 10 ng/mL human recombinant stem cell factor). Cells were plated out at a density of 1.times.10.sup.4 cells/mL in duplicates in 6-well microplates with test antibody at concentrations ranging from 0.1-100 ng/m L. Plates were incubated at 37.degree. C. and 5% CO.sub.2 for two weeks. Duplicate samples from each well were recovered for cell counts and staining with benzidine (Reference Fibach, E., 1998 Hemoglobin, 22:5-6, 445-458).

[0197] The results are shown in FIG. 9. Specifically, Ab198 induced the proliferation of human erythroid producing cells derived from progenitor cells in a dose dependent manner. The number of proliferating cells and the percentage expressing hemoglobin, as indicated by staining with benzidine, was reduced compared to the EPO treated controls again probably due to a reduced proliferative signal.

Example 7

Cynomolgus Bone Marrow CFUe Assay

[0198] Bone marrow was harvested from cynomolgus monkeys and diluted 1:2 with PBS. Three ml of the diluted bone marrow was layered over six ml of Lymphoprep (Gibco (Invitrogen), Carlsbad, Calif. Cat. #1001967), centrifuged at 2700 rpm for 20 minutes and the buffy coat recovered and diluted in 10 ml IMDM-2% FBS. Cells were centrifuged and resuspended at 10.sup.6 cells/ml in IMDM-2% FBS. Cells (0.3 ml) were added to tubes containing 2.4 ml Methocult (StemCell Technologies, Vancouver, Canada) Cat. #04230), 0.3 ml stem cell growth factor (Sigma, Cat. #S7901, 100 ug/ml), 0.3 ml EPO(R& D Systems, Minneapolis, Minn.), test antibody (Ab198), or IMDM-2% FBS. After mixing, 1.1 ml of the Methocult suspension was added to a 35 mm non tissue culture treated sterile petri dish and incubated at 37.degree. C., 5% CO.sub.2 for 2 weeks. Colonies were identified microscopically. The results of this assay are shown in FIG. 10 demonstrate that Ab198 induced the formation of CFU-E colonies (although the number of colonies was reduced compared to that observed with the EPO control).

Example 8

ELISA to Measure Binding of SE-3 Peptide

[0199] 96 well polystyrene plates (Dynatec (Elk Grove Village, Ill.) Immunolon 4) were coated with 80 ul of 5 ug/ml soluble EPO receptor (sEPOR) (R&D Systems (Minneapolis, Minn.) Cat. #307-ER/LF)--, or peptide SE-3 (PGNYSFSYQLEDEPWKLCRLHWAPTARGAV) (described in U.S. Pat. No. 6,319,499) diluted in 0.015M Na.sub.2CO.sub.3, 0.035M NaHCO.sub.3, pH 9.4 for 2 hours at room temperature and overnight at 4.degree. C. Plates were blocked for 30 minutes at room temperature with 100 ul of 5% BSA in PBS (Gibco (Invitrogen (Carlsbad, Calif.)) Cat. #10010). After removal of blocking solution, 50 ul of Ab12 at 5 ug/ml in PBS with 1% BSA was added to wells and plates were incubated at room temperature for 2 hours. Plates were washed three times using a Skatron 400 Plate Washer with PBS/0.05% Tween 20 and 50 ul of secondary antibody diluted in PBS/0.25% BSA/0.05% Tween 20 added to the wells. For Abl 2, goat anti-human IgG (Fc)-HRP (Caltag (Burlingame, Calif.) Cat. #H10507) diluted 1:1000 was used and for Ab 71A (available from the American Type Culture Collection HB11689, also described in U.S. Pat. No. 6,319,499), goat anti mouse IgG (Fc)-HRP (Jackson Laboratories (West Grove, Pa.) Cat. #115-035-164) diluted 1:5000 was used. After a 1 hour incubation at room temperature, plates were washed three times as before and 50 ul of OPD Developing Reagent (Sigma #P9187) added to each well. Color development was stopped by addition of 50 ul of 1N HCl to the wells and optical density measured at 490 nm on a Victor 1420 Multi-Label Counter.

[0200] FIG. 11 shows that Ab12 does not interact (i.e. bind) with SE-3 peptide. Ab 71A does interact (i.e. binds) with the SE-3 peptide Both Abs 12, and 71A interacted with immobilized erythropoietin receptor.

Example 9

EPO Dependent Proliferation Assay

[0201] Primary hybridoma supernatants were diluted in assay medium and tested for their ability to stimulate the proliferation of the F36E human erythroleukemic cells as described in EXAMPLE 5. Results with five primary supernatants are shown in FIG. 12. These samples stimulated the proliferation of F36E cells.

Example 10

ELISA to Measure Binding of Hybridoma Supernatants to SE-3 Peptide

[0202] Forty-two primary hybridoma supernatants were tested for their ability to bind to either immobilized EPO receptor or peptide SE-3 as described in EXAMPLE 10. FIG. 13 shows that whereas all the hybridoma supernatants tested interact with immobilized EPO receptor, only sample 16 interacted with SE-3 peptide at levels above background.

Example 11

Comparison of Erythropoietic Activity of Gamma-1 Ab12 Versus Gamma-2 Ab12

[0203] Proliferation assays (as described in Example 4) were performed to compare the erythropoietic activity of gamma-1 Ab 12 and gamma-2 Ab 12 on F36e human erythroleukemic cells. The results are shown in FIG. 31. As FIG. 31 shows, gamma-2 Ab 12 was more effective at stimulating proliferation of the F36E cell line than gamma-1 Ab 12.

Example 12

Effect of Ab 12 on Erythropoiesis In Vivo

[0204] (a) Construction of mEpoR-/-, hEopR+transgenic mice: Transgenic mice that produced only human EpoR (hEpoR+, single allele) and no endogenous mouse EpoR (mEpoR-/-, double allele mutation) were generated as described in Liu. C. et al., Journal of Biological Chemistry 272:32395 (1997) and Yu, X., et al., Blood, 98(2):475 (2001). Breeding colonies were established to generate mice for in vivo studies of eryhthropoiesis.

[0205] (b) Multiple dosing regimen: In initial experiments, animals were subjected to a multiple dosing regimen of Ab 12 to determine whether the antibody would cause an increase in reticulocyte counts and/or % hematocrit. Five transgenic mice (mEpoR-/-, hEpoR+, were injected subcutaneously with either 5 .mu.g or 50 .mu.g of Ab 12 in 0.2 mL vehicle (phosphate buffered saline [PBS] containing 0.1% bovine serum albumin ([BSA]). Control animals also were injected in the same manner with equal volumes of the vehicle alone or vehicle containing 5 U Epogen.RTM. (Amgen.RTM., Thousand Oaks, Calif.). All animals were dosed over a three-week period in accordance with the following schedule:

TABLE-US-00007 Week 1 Week 2 Week 3 Monday, Tuesday, Wednesday, Monday, Wednesday, Monday, Friday Friday Wednesday

Sample bleeds were taken on day 4 (Thursday of week 1) for determining reticulocyte counts and on day 19 (Friday of week 3) for determining hematocrits. Reticulocyte counts and hematocrit determinations were made using methods well known in the art. As FIG. 32 shows, Ab 12 caused a statistically significant increase (over controls) in reticulocyte count and % hematocrit in animals receiving either 5 or 50 .mu.g of Ab 12 antibody.

[0206] (c) Weekly dosing regimen: To assess whether the results seen under a multiple dosing regimen still would be observed in animals receiving fewer doses of Ab 12, transgenic mice were injected (as described in (b) above) with varying concentrations (0.5, 2.5, 5.0, 50 and 250 .mu.g) of Ab 12 or a control, Aranesp.TM. (Amgen.RTM., Thousand Oaks, Calif.), a more active variant of Epogen.RTM. on days 1, 8 and 15 and bled on days 4 and 19 for determination of reticulocyte count and hematocrit, respectively. Control animals received a single dose of vehicle only or a human IgG2 isotype control. FIG. 33 shows that Ab 12 caused a statistically significant increase (over vehicle and isotype controls) in percent hematocrit with all but the lowest concentrations tested.

[0207] (d) Single versus weekly dosing regimens: To determine whether a single dose of Ab-12 would have an effect on erythropoiesis after 3 weeks, transgenic mice were dosed with Ab 12 (50 .mu.g), at one week intervals for 3 weeks or with a single dose of Ab 12 (150 .mu.g) and bled on day 19 for determination of percent hematocrit. Control animals received vehicle alone, a single dose of Aranesp.TM. (900 ng) or 3 total doses of Aranesp.TM. injected at weekly intervals (300 ng.times.3). FIG. 34 shows that both dosing regimens of Ab 12 caused a statistically significant increase in percent hematocrit over the vehicle control. In contrast, the single dose regimen of Aranesp.TM. did not have this effect.

[0208] All abstracts, references, patents and published patent applications referred to herein are hereby incorporated by reference.

[0209] The present invention is illustrated by way of the foregoing description and examples. The foregoing description is intended as a non-limiting illustration, since many variations will become apparent to those skilled in the art in view thereof.

[0210] Changes can be made to the composition, operation and arrangement of the method of the present invention described herein without departing from the concept and scope of the invention.

Sequence CWU 1

1

115130PRTHomo sapiens 1Pro Gly Asn Tyr Ser Phe Ser Tyr Gln Leu Glu Asp Glu Pro Trp Lys1 5 10 15Leu Cys Arg Leu His Gln Ala Pro Thr Ala Arg Gly Ala Val 20 25 302349DNAHomo sapiens 2caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgc ctccatcagt agttactact ggagctggat ccggcagccc 120ccagggaagg gactggagtg gattgggtat atctattaca gtgggagcac caactacaac 180ccctccctca agagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctg 240aagctgaggt ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgag agagcgactg 300gggatcgggg actactgggg ccaaggaacc ctggtcaccg tctcctcag 3493116PRTHomo sapiens 3Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ala Ser Ile Ser Ser Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Arg Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Glu Arg Leu Gly Ile Gly Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110Thr Val Ser Ser 1154322DNAHomo sapiens 4gacatccagc tgacccaatc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatactt accctccgac gttcggccaa 300gggaccaagg tggaaatcaa ac 3225107PRTHomo sapiens 5Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Thr Tyr Pro Pro 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 1056370DNAHomo sapiens 6caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgtag cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agttgaggac acggctgtgt attactgtgc gagagatcac 300ggtgggaggt acgtctacga ctacggtatg gacgtctggg gccaagggac cacggtcacc 360gtctcctcag 3707123PRTHomo sapiens 7Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp His Gly Gly Arg Tyr Val Tyr Asp Tyr Gly Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 1208322DNAHomo sapiens 8gacatccaga tgacccaatc tccatcttcc gtgtctgcat ctataggaga cagagtctcc 60atcacttgtc gggcgagtca gggtattagc agctggttag cctggtatca gcagaaacca 120gggaaagccc ctacgctcct tatctatgct gcatccactt tgcaacgtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 300gggaccaaag tggatatcaa ac 3229107PRTHomo sapiens 9Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Ile Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Thr Leu Leu Ile 35 40 45Tyr Ala Ala Ser Thr Leu Gln Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 10510370DNAHomo sapiens 10caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agttgaggac acggctgtgt attactgtgc gagagatcac 300ggtgggaggt acgtctacga ctacggtatg gacgtctggg gccaagggac cacggtcacc 360gtctcctcag 37011123PRTHomo sapiens 11Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp His Gly Gly Arg Tyr Val Tyr Asp Tyr Gly Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 12012322DNAHomo sapiens 12gacatccaga tgacccaatc tccatcttcc gtgtctgcat ctgtaggaga cagagtctcc 60atcacttgtc gggcgagtca gggtattagc agctggttag tctggtatca gcagaaacca 120gggaaagccc ctgcgctcct aatctatgct gcatccagtt tgcagcgtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagac ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 300gggaccaaag tggatatcaa ac 32213107PRTHomo sapiens 13Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Val Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Ala Leu Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 10514370DNAHomo sapiens 14caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggtagtt atatcatatg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agttgaggac acggctgtgt attactgtgc gagagatcac 300ggtgggaggt acgtctacga ctacggtatg gacgtctggg gccaagggac cacggtcacc 360gtctcctcag 37015123PRTHomo sapiens 15Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Val Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp His Gly Gly Arg Tyr Val Tyr Asp Tyr Gly Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 12016322DNAHomo sapiens 16gacatccaga tgacccaatc tccatcttcc gtgtctgcat ctgtaggaga cagagtctcc 60atcacttgtc gggcgagtca gggtattagc agctggttag cctggtatca gcagaaacca 120gggaaagccc ctacgctcct aatctatgct gcatccagtt tgcaacgtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 300gggaccaaag tggatatcaa ac 32217107PRTHomo sapiens 17Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Thr Leu Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 10518349DNAHomo sapiens 18caggtgcagc tggtggagtc ggggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt aaatatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt ttatggtatg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc gagaggtccg 300tactactttg actactgggg ccagggaacc ctggtcaccg tctcctcag 34919116PRTHomo sapiens 19Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Lys Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Leu Trp Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Pro Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110Thr Val Ser Ser 11520325DNAHomo sapiens 20gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac tggcatccca 180gacaggttca gtggcagtgg gtctgggaca gacttcactg tcaccatcag cagactggaa 240cctgaagatt ttgcagtgta ttactgtcag cagtatggta gttcaccgtg gacgttcggc 300caagggacca aggtggaaat caaac 32521108PRTHomo sapiens 21Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Val Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 10522322DNAHomo sapiens 22gacatccaga tgacccaatc tccatcttcc gtgtccgcat ctgtaggaga cagagtctcc 60atcacttgtc gggcgagtca gggtattagc agctggttag cctggtatca gcagaaacca 120gggaaagccc ctacgctcct aatctatgct gcatccagtt tgcaacgtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 300gggaccaaag tggatatcaa ac 32223107PRTHomo sapiens 23Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Thr Leu Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 10524322DNAHomo sapiens 24gacatccaga tgacccaatc tccatcttcc gtgtctgcat ctgtaggaga cagagtctcc 60atcacttgtc gggcgagtca gggtattagc agctggttag cctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaacgtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 300gggaccaaag tggatatcaa ac 32225107PRTHomo sapiens 25Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 10526322DNAHomo sapiens 26gacatccaga tgacccagtc tccatcttcc gtgtctacat ctgtaggaga cagagtctcc 60atcacttgtc gggcgagtca gggtattggc agctggttag cctggtatca gcagaaacca 120gggcaagccc ctacgctcct aatctatgct gcatccagtt tgcaacgtgg ggtcccatca 180agattcagcg gcagtggatc tgggacagat ttcactctca ccatcaacag cctgcagcct 240gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 300gggaccaaag tggatgtcaa ac 32227107PRTHomo sapiens 27Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Thr Ser Val Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Gly Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Thr Leu Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Val Lys 100 10528322DNAHomo sapiens 28gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtctcc 60atcacttgtc gggcgagtca gggtattggc agctggttag cctggtatca gcagaaacca 120gggcaagccc ctacgctcct aatctatgct gcatccagtt tgcaacgtgg ggtcccatca 180agattcagcg gcagtggatc tgggacagat ttcactctca ccatcaacag cctgcagcct 240gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 300gggaccaaag tggatgtcaa ac 32229107PRTHomo sapiens 29Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Gly Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Thr Leu Leu Ile 35

40 45Tyr Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Val Lys 100 10530349DNAHomo sapiens 30caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatggtttg atggaaataa taaattctat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agtcgaggac acggctgtgt attactgtgc gcgaggcggg 300agctactggg actactgggg ccagggaacc ctggtcaccg tctcctcag 34931116PRTHomo sapiens 31Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Phe Asp Gly Asn Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Tyr Trp Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110Thr Val Ser Ser 11532336DNAHomo sapiens 32gatattgtga tgacccagac tccactcttc tcatttgtca tgattggaca gccggcctcc 60atctcctgca ggtctaggca aagcctcgta cacagtgatg gaaacaccta cttgaattgg 120cttcagcaga ggccaggcca gcctccaaga ctcctaattt ataagacttc taaccggttc 180tctggggtcc cagatagatt cagtggcagt ggggcaggga cagatttcac actgaaaatc 240agcagggtgg aagctgagga tgtcggggtt tattactgta tgcaagctac acaatttcct 300atcacgttcg gccaagggac acgactggag attaaa 33633112PRTHomo sapiens 33Asp Ile Val Met Thr Gln Thr Pro Leu Phe Ser Phe Val Met Ile Gly1 5 10 15Gln Pro Ala Ser Ile Ser Cys Arg Ser Arg Gln Ser Leu Val His Ser 20 25 30Asp Gly Asn Thr Tyr Leu Asn Trp Leu Gln Gln Arg Pro Gly Gln Pro 35 40 45Pro Arg Leu Leu Ile Tyr Lys Thr Ser Asn Arg Phe Ser Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ala Gly Thr Asp Phe Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala 85 90 95Thr Gln Phe Pro Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100 105 11034370DNAHomo sapiens 34caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt agctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatcatatg atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agttgaggac acggctgtgt attactgtgc gaaagatcac 300ggtgggaggt acgtctacga ctacggtatg gacgtctggg gccaagggac cacggtcacc 360gtctcctcag 37035123PRTHomo sapiens 35Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Asp His Gly Gly Arg Tyr Val Tyr Asp Tyr Gly Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 12036322DNAHomo sapiens 36gacatccaga tgacccagtc tccatcttcc gtgtctgcat ctgtaggaga cagagtctcc 60atcacttgtc gggcgagtca gggtattggc agctggttag cctggtatca gcagaaacca 120gggcaagccc ctacgctcct aatctatgct gcctccagtt tgcaacgtgg ggtcccatca 180agattcagcg gcagtggatc tgggacagat ttcactctca ccatcaacag cctgcagcct 240gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 300gggaccaaag tggatgtcaa ac 32237107PRTHomo sapiens 37Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly Ile Gly Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Thr Leu Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Arg Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Asn Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn Ser Phe Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Val Lys 100 10538348DNAHomo sapiens 38caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgc ctccatcagt aattactact ggagctggat ccggcagccc 120ccagggaagg gactggagtg gattgggtat gtctcttaca gtgggagtac gtactacaac 180ccctccctca agggtcgagt caccatgtca gtagacacgt ccaagaacca gttctccctg 240aagctgagct ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgag agaaaaactg 300gggattggag actactgggg ccagggaacc ctggtcaccg tctcctca 34839116PRTHomo sapiens 39Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ala Ser Ile Ser Asn Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Val Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Gly Arg Val Thr Met Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Glu Lys Leu Gly Ile Gly Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110Thr Val Ser Ser 11540322DNAHomo sapiens 40gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattaaa aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatagtt atccgtgcag ttttggccag 300gggaccaagc tggagatcaa ac 32241107PRTHomo sapiens 41Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Lys Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Cys 85 90 95Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10542354DNAHomo sapiens 42caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcacagac cctgtccctc 60acctgcactg tctctggtgc ctccatcagc agtggtgctt actactggag ttggatccgc 120cagcacccag ggaagggcct ggagtggatt gggtacatct ataagagtga gacctcctac 180tacaacccgt ccctcaagag tcgacttacc ctatcagtag acacgtctaa gaaccagttc 240tccctgaacc tgatctctgt gactgccgcg gacacggccg tgtattattg tgcgagagat 300aaactgggga tcgcggacta ctggggccag ggaaccctgg tcaccgtctc ctca 35443118PRTHomo sapiens 43Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Gln1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ala Ser Ile Ser Ser Gly 20 25 30Ala Tyr Tyr Trp Ser Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu 35 40 45Trp Ile Gly Tyr Ile Tyr Lys Ser Glu Thr Ser Tyr Tyr Asn Pro Ser 50 55 60Leu Lys Ser Arg Leu Thr Leu Ser Val Asp Thr Ser Lys Asn Gln Phe65 70 75 80Ser Leu Asn Leu Ile Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95Cys Ala Arg Asp Lys Leu Gly Ile Ala Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11544322DNAHomo sapiens 44gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca ggacattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccaatt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatagct accctcccac tttcggcgga 300gggaccaagg tggaaatcaa ac 32245107PRTHomo sapiens 45Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Asn Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Pro 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 10546349DNAHomo sapiens 46caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgt ctccatcagt aattactact ggagctggat ccggcagtcc 120ccagggaagg gactggagtg gattggatat atctattaca gtgggagtcc ctattacaac 180ccctccctca agagtcgagt cactatatct gcagacacgt ccaagaacca attctccctg 240aagctgagct ctgtgaccgc tgcggacacg gccatttatt actgtgcgag agaaaaactg 300gggattggag actactgggg ccagggaacc ctggtcaccg tctcctcag 34947116PRTHomo sapiens 47Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val Ser Ile Ser Asn Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Tyr Tyr Ser Gly Ser Pro Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Ala Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Ile Tyr Tyr Cys Ala 85 90 95Arg Glu Lys Leu Gly Ile Gly Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110Thr Val Ser Ser 11548322DNAHomo sapiens 48gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtcggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatagtt accctcccac tttcggccct 300gggaccaagg tggatatcaa ac 32249107PRTHomo sapiens 49Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Pro 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100 10550349DNAHomo sapiens 50caggtgcagc tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtgg ctccatcagt cgttactact ggagctggat ccggcagccc 120ccagggaagg gactggagtg gattgggtat gtctcttaca gtgggagcac ctactacaac 180ccctccctca agagtcgagt caccatatca gtagacacgt ccaagaacca gttctccctg 240aagctgagct ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgag agataaactg 300gggattggag actactgggg ccagggaacc ctggtcaccg tctcctcag 34951116PRTHomo sapiens 51Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Arg Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Tyr Val Ser Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Asp Lys Leu Gly Ile Gly Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110Thr Val Ser Ser 11552322DNAHomo sapiens 52gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaaccg 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatagtt acccgtgcag ttttggccag 300gggaccaagc tggagatcaa ac 32253107PRTHomo sapiens 53Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Cys 85 90 95Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10554355DNAHomo sapiens 54caggtgcagc tgcaggagtc gggcccagga ctggtgaagc ctttacagac cctgtccctc 60acctgcactg tctctggtgg ctccatcagc agtggtgttt actactggag ctggatccgc 120cagcacccag ggaagggcct ggagtggatt gggtacatct ataacagtaa gacctcctat 180tataatccgt ccctcaagag tcgacttacc ctatcagtag acacgtctaa gaaccagttc 240tccctgaacc tgatctctgt gactgccgcg gacacggccg tgtattactg tgcgagagat 300aaattgggga tcgcggacta ctggggccag ggaaccctgg tcaccgtctc ctcag 35555118PRTHomo sapiens 55Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Leu Gln1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Gly 20 25 30Val Tyr Tyr Trp Ser Trp Ile Arg Gln His Pro Gly Lys Gly Leu Glu 35 40 45Trp Ile Gly Tyr Ile Tyr Asn Ser Lys Thr Ser Tyr Tyr Asn Pro Ser 50 55 60Leu Lys Ser Arg Leu Thr Leu Ser Val Asp Thr Ser Lys Asn Gln Phe65 70 75 80Ser Leu Asn Leu Ile Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95Cys Ala Arg Asp Lys Leu Gly Ile Ala Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser 11556322DNAHomo sapiens 56gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc ggacaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 120gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg caacttatta ctgtctacag cataatagct accctcccac tttcggcgga 300gggaccaagg tggagatcaa ac 32257107PRTHomo sapiens 57Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5

10 15Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Gly Ile Arg Asn Asp 20 25 30Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Arg Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn Ser Tyr Pro Pro 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100 1055834PRTHomo sapiens 58Gly Ala Ser Ile Ser Ser Tyr Tyr Trp Ser Tyr Ile Tyr Tyr Ser Gly1 5 10 15Ser Thr Asn Tyr Asn Pro Ser Leu Lys Ser Glu Arg Leu Gly Ile Gly 20 25 30Asp Tyr5941PRTHomo sapiens 59Gly Phe Thr Phe Ser Ser Tyr Gly Met His Val Ile Ser Tyr Asp Gly1 5 10 15Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys Gly Asp His Gly Gly Arg 20 25 30Tyr Val Tyr Asp Tyr Gly Met Asp Val 35 406034PRTHomo sapiens 60Gly Phe Thr Phe Ser Lys Tyr Gly Met His Val Leu Trp Tyr Asp Gly1 5 10 15Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys Gly Asp Gly His Tyr Phe 20 25 30Asp Tyr6134PRTHomo sapiens 61Gly Phe Thr Phe Ser Ser Tyr Gly Met His Val Ile Trp Phe Asp Gly1 5 10 15Asn Asn Lys Phe Tyr Ala Asp Ser Val Lys Gly Ala Pro Ala Tyr Trp 20 25 30Asp Tyr6227PRTHomo sapiens 62Arg Ala Ser Gln Gly Ile Arg Asn Asp Leu Gly Ala Ala Ser Ser Leu1 5 10 15Gln Ser Leu Gln His Asn Thr Tyr Pro Pro Thr 20 256327PRTHomo sapiens 63Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala Ala Ala Ser Thr Leu1 5 10 15Gln Arg Gln Gln Ala Asn Ser Phe Pro Phe Thr 20 256429PRTHomo sapiens 64Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Val Ala Leu Ala Ala Ser1 5 10 15Ser Leu Gln Arg Gln Gln Ala Asn Ser Phe Pro Phe Thr 20 256527PRTHomo sapiens 65Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala Ala Ala Ser Ser Leu1 5 10 15Gln Arg Gln Gln Ala Asn Ser Phe Pro Phe Thr 20 256627PRTHomo sapiens 66Arg Ala Ser Gln Gly Ile Gly Ser Trp Leu Ala Ala Ala Ser Ser Leu1 5 10 15Gln Arg Gln Gln Ala Asn Ser Phe Pro Phe Thr 20 256732PRTHomo sapiens 67Arg Ser Arg Gln Ser Leu Val His Ser Asp Gly Asn Thr Tyr Leu Asn1 5 10 15Lys Thr Ser Asn Arg Phe Ser Met Gln Ala Thr Gln Phe Pro Ile Thr 20 25 306828PRTHomo sapiens 68Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala Gly Ala Ser Ser1 5 10 15Arg Ala Thr Gln Gln Tyr Gly Ser Ser Pro Trp Thr 20 25691990DNAHomo sapiens 69atgaagcatc tgtggttctt ccttctccta gtggcagctc ccagatgggt cctgtcccag 60gtgcagctgc aggagtcggg cccaggactg gtgaagcctt cggagaccct gtccctcacc 120tgcactgtct ctggtgcctc catcagtagt tactactgga gctggatccg gcagccccca 180gggaagggac tggagtggat tgggtatatc tattacagtg ggagcaccaa ctacaacccc 240tccctcaaga gtcgagtcac catatcagta gacacgtcca agaaccagtt ctccctgaag 300ctgaggtctg tgaccgctgc ggacacggcc gtgtattact gtgcgagaga gcgactgggg 360atcggggact actggggcca aggaaccctg gtcaccgtct cctcagcctc caccaagggc 420ccatcggtct tccccctggc gccctgctct agaagcacct ccgagagcac agccgccctg 480ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgct 540ctgaccagcg gcgtgcacac cttcccagct gtcctacagt cctcaggact ctactccctc 600agcagcgtgg tgaccgtgcc ctccagcaac ttcggcaccc agacctacac ctgcaacgta 660gatcacaagc ccagcaacac caaggtggac aagacagttg gtgagaggcc agctcaggga 720gggagggtgt ctgctggaag ccaggctcag ccctcctgcc tggacgcacc ccggctgtgc 780agccccagcc cagggcagca aggcaggccc catctgtctc ctcacccgga ggcctctgcc 840cgccccactc atgctcaggg agagggtctt ctggcttttt ccaccaggct ccaggcaggc 900acaggctggg tgcccctacc ccaggccctt cacacacagg ggcaggtgct tggctcagac 960ctgccaaaag ccatatccgg gaggaccctg cccctgacct aagccgaccc caaaggccaa 1020actgtccact ccctcagctc ggacaccttc tctcctccca gatccgagta actcccaatc 1080ttctctctgc agagcgcaaa tgttgtgtcg agtgcccacc gtgcccaggt aagccagccc 1140aggcctcgcc ctccagctca aggcgggaca ggtgccctag agtagcctgc atccagggac 1200aggccccagc tgggtgctga cacgtccacc tccatctctt cctcagcacc acctgtggca 1260ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc 1320cctgaggtca cgtgcgtggt ggtggacgtg agccacgaag accccgaggt ccagttcaac 1380tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccacggga ggagcagttc 1440aacagcacgt tccgtgtggt cagcgtcctc accgttgtgc accaggactg gctgaacggc 1500aaggagtaca agtgcaaggt ctccaacaaa ggcctcccag cccccatcga gaaaaccatc 1560tccaaaacca aaggtgggac ccgcggggta tgagggccac atggacagag gccggctcgg 1620cccaccctct gccctgggag tgaccgctgt gccaacctct gtccctacag ggcagccccg 1680agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag 1740cctgacctgc ctggtcaaag gcttctaccc cagcgacatc gccgtggagt gggagagcaa 1800tgggcagccg gagaacaact acaagaccac acctcccatg ctggactccg acggctcctt 1860cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc 1920atgctccgtg atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc 1980tccgggtaaa 1990701990DNAHomo sapiens 70tttacccgga gacagggaga ggctcttctg cgtgtagtgg ttgtgcagag cctcatgcat 60cacggagcat gagaagacgt tcccctgctg ccacctgctc ttgtccacgg tgagcttgct 120gtagaggaag aaggagccgt cggagtccag catgggaggt gtggtcttgt agttgttctc 180cggctgccca ttgctctccc actccacggc gatgtcgctg gggtagaagc ctttgaccag 240gcaggtcagg ctgacctggt tcttggtcat ctcctcccgg gatgggggca gggtgtacac 300ctgtggttct cggggctgcc ctgtagggac agaggttggc acagcggtca ctcccagggc 360agagggtggg ccgagccggc ctctgtccat gtggccctca taccccgcgg gtcccacctt 420tggttttgga gatggttttc tcgatggggg ctgggaggcc tttgttggag accttgcact 480tgtactcctt gccgttcagc cagtcctggt gcacaacggt gaggacgctg accacacgga 540acgtgctgtt gaactgctcc tcccgtggct ttgtcttggc attatgcacc tccacgccgt 600ccacgtacca gttgaactgg acctcggggt cttcgtggct cacgtccacc accacgcacg 660tgacctcagg ggtccgggag atcatgaggg tgtccttggg ttttgggggg aagaggaaga 720ctgacggtcc tgccacaggt ggtgctgagg aagagatgga ggtggacgtg tcagcaccca 780gctggggcct gtccctggat gcaggctact ctagggcacc tgtcccgcct tgagctggag 840ggcgaggcct gggctggctt acctgggcac ggtgggcact cgacacaaca tttgcgctct 900gcagagagaa gattgggagt tactcggatc tgggaggaga gaaggtgtcc gagctgaggg 960agtggacagt ttggcctttg gggtcggctt aggtcagggg cagggtcctc ccggatatgg 1020cttttggcag gtctgagcca agcacctgcc cctgtgtgtg aagggcctgg ggtaggggca 1080cccagcctgt gcctgcctgg agcctggtgg aaaaagccag aagaccctct ccctgagcat 1140gagtggggcg ggcagaggcc tccgggtgag gagacagatg gggcctgcct tgctgccctg 1200ggctggggct gcacagccgg ggtgcgtcca ggcaggaggg ctgagcctgg cttccagcag 1260acaccctccc tccctgagct ggcctctcac caactgtctt gtccaccttg gtgttgctgg 1320gcttgtgatc tacgttgcag gtgtaggtct gggtgccgaa gttgctggag ggcacggtca 1380ccacgctgct gagggagtag agtcctgagg actgtaggac agctgggaag gtgtgcacgc 1440cgctggtcag agcgcctgag ttccacgaca ccgtcaccgg ttcggggaag tagtccttga 1500ccaggcagcc cagggcggct gtgctctcgg aggtgcttct agagcagggc gccaggggga 1560agaccgatgg gcccttggtg gaggctgagg agacggtgac cagggttcct tggccccagt 1620agtccccgat ccccagtcgc tctctcgcac agtaatacac ggccgtgtcc gcagcggtca 1680cagacctcag cttcagggag aactggttct tggacgtgtc tactgatatg gtgactcgac 1740tcttgaggga ggggttgtag ttggtgctcc cactgtaata gatataccca atccactcca 1800gtcccttccc tgggggctgc cggatccagc tccagtagta actactgatg gaggcaccag 1860agacagtgca ggtgagggac agggtctccg aaggcttcac cagtcctggg cccgactcct 1920gcagctgcac ctgggacagg acccatctgg gagctgccac taggagaagg aagaaccaca 1980gatgcttcat 199071241PRTHomo sapiens 71Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Leu Ala Ala Pro1 5 10 15Arg Trp Val Leu Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu 20 25 30Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ala 35 40 45Ser Ile Ser Ser Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys 50 55 60Gly Leu Glu Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr65 70 75 80Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Ala Ser Pro Thr 85 90 95Ser Lys Asn Gln Phe Ser Leu Lys Leu Arg Ser Val Thr Ala Ala Asp 100 105 110Thr Ala Val Tyr Tyr Cys Ala Arg Glu Arg Leu Gly Ile Gly Asp Tyr 115 120 125Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 130 135 140Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser145 150 155 160Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 165 170 175Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 180 185 190Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 195 200 205Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 210 215 220Ala Ser Pro His Lys Pro Ser Asn Thr Lys Val Ala Ser Pro Lys Thr225 230 235 240Val7212PRTHomo sapiens 72Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro1 5 1073115PRTHomo sapiens 73Ala Pro Pro Val Ala Leu Ala Gly Pro Ser Val Phe Leu Phe Pro Pro1 5 10 15Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 20 25 30Val Val Val Ala Ser Pro Val Ser His Glu Asp Pro Glu Val Gln Phe 35 40 45Asn Trp Tyr Val Ala Ser Pro Gly Val Glu Val His Asn Ala Lys Thr 50 55 60Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val65 70 75 80Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 85 90 95Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser 100 105 110Lys Thr Lys 11574107PRTHomo sapiens 74Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu1 5 10 15Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 20 25 30Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 35 40 45Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe 50 55 60Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly65 70 75 80Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 85 90 95Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 100 10575310PRTHomo sapiens 75Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr1 5 10 15Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 20 25 30Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 35 40 45Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr 50 55 60Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys65 70 75 80Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro 85 90 95Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 100 105 110Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 115 120 125Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly 130 135 140Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn145 150 155 160Ser Thr Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp 165 170 175Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro 180 185 190Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu 195 200 205Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 210 215 220Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile225 230 235 240Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 245 250 255Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 260 265 270Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 275 280 285Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 290 295 300Ser Leu Ser Pro Gly Lys305 31076552DNAHomo sapiens 76atgagggtcc ccgctcagct cctggggctc ctgctgctct ggttcccagg tgccaggtgt 60aagcttgaca tccagctgac ccaatctcca tcctccctgt ctgcatctgt aggagacaga 120gtcaccatca cttgccgggc aagtcagggc attagaaatg atttaggctg gtatcagcag 180aaaccaggga aagcccctaa gcgcctgatc tatgctgcat ccagtttgca aagtggggtc 240ccatcaaggt tcagcggcag tggatctggg acagaattca ctctcacaat cagcagcctg 300cagcctgaag attttgcaac ttattactgt ctacagcata atacttaccc tccgacgttc 360ggccaaggga ccaaggtgga aatcaaacga actgtggctg caccatctgt cttcatcttc 420ccgccatctg atgagcagtt gaaatctgga actgctagcg ttgtgtgcct gctgaataac 480ttctatccca gagaggccaa agtacagtgg aaggtggata acgccctcca atcgggtaac 540tcccaggaga gt 55277552DNAHomo sapiens 77actctcctgg gagttacccg attggagggc gttatccacc ttccactgta ctttggcctc 60tctgggatag aagttattca gcaggcacac aacgctagca gttccagatt tcaactgctc 120atcagatggc gggaagatga agacagatgg tgcagccaca gttcgtttga tttccacctt 180ggtcccttgg ccgaacgtcg gagggtaagt attatgctgt agacagtaat aagttgcaaa 240atcttcaggc tgcaggctgc tgattgtgag agtgaattct gtcccagatc cactgccgct 300gaaccttgat gggaccccac tttgcaaact ggatgcagca tagatcaggc gcttaggggc 360tttccctggt ttctgctgat accagcctaa atcatttcta atgccctgac ttgcccggca 420agtgatggtg actctgtctc ctacagatgc agacagggag gatggagatt gggtcagctg 480gatgtcaagc ttacacctgg cacctgggaa ccagagcagc aggagcccca ggagctgagc 540ggggaccctc at 55278184PRTHomo sapiens 78Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro1 5 10 15Gly Ala Arg Cys Lys Leu Asp Ile Gln Leu Thr Gln Ser Pro Ser Ser 20 25 30Leu Ser Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser 35 40 45Gln Gly Ile Arg Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys 50 55 60Ala Pro Lys Arg Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val65 70 75 80Pro Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr 85 90 95Ile Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln 100 105 110His Asn Thr Tyr Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 115 120 125Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp 130 135 140Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn145 150 155 160Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu 165 170 175Gln Ser Gly Asn Ser Gln Glu Ser 1807931PRTHomo sapiens 79Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln1 5 10 15Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser 20 25 30802011DNAHomo sapiens 80atggaattgg ggctccgctg ggttttcctc gttgctcttt taagaggtgt ccagtgtcag 60gtgcagctgg tggagtctgg gggaggcgtg gtccagcctg ggaggtccct gagactctcc 120tgtgtagcct ctggattcac cttcagtagc tatggcatgc actgggtccg ccaggctcca 180ggcaaggggc tggagtgggt ggcagttata tcatatgatg gaagtaataa atactatgca 240gactccgtga agggccgatt caccatctcc agagacaatt ccaagaacac gctgtatctg 300caaatgaaca gcctgagagt tgaggacacg gctgtgtatt actgtgcgag agatcacggt 360gggaggtacg tctacgacta cggtatggac gtctggggcc aagggaccac ggtcaccgtc 420tcctcagcct ccaccaaggg cccatcggtc ttccccctgg cgccctgctc tagaagcacc 480tccgagagca cagccgccct gggctgcctg gtcaaggact acttccccga accggtgacg 540gtgtcgtgga actcaggcgc tctgaccagc ggcgtgcaca ccttcccagc tgtcctacag 600tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcaa cttcggcacc

660cagacctaca cctgcaacgt agatcacaag cccagcaaca ccaaggtgga caagacagtt 720ggtgagaggc cagctcaggg agggagggtg tctgctggaa gccaggctca gccctcctgc 780ctggacgcac cccggctgtg cagccccagc ccagggcagc aaggcaggcc ccatctgtct 840cctcacccgg aggcctctgc ccgccccact catgctcagg gagagggtct tctggctttt 900tccaccaggc tccaggcagg cacaggctgg gtgcccctac cccaggccct tcacacacag 960gggcaggtgc ttggctcaga cctgccaaaa gccatatccg ggaggaccct gcccctgacc 1020taagccgacc ccaaaggcca aactgtccac tccctcagct cggacacctt ctctcctccc 1080agatccgagt aactcccaat cttctctctg cagagcgcaa atgttgtgtc gagtgcccac 1140cgtgcccagg taagccagcc caggcctcgc cctccagctc aaggcgggac aggtgcccta 1200gagtagcctg catccaggga caggccccag ctgggtgctg acacgtccac ctccatctct 1260tcctcagcac cacctgtggc aggaccgtca gtcttcctct tccccccaaa acccaaggac 1320accctcatga tctcccggac ccctgaggtc acgtgcgtgg tggtggacgt gagccacgaa 1380gaccccgagg tccagttcaa ctggtacgtg gacggcgtgg aggtgcataa tgccaagaca 1440aagccacggg aggagcagtt caacagcacg ttccgtgtgg tcagcgtcct caccgttgtg 1500caccaggact ggctgaacgg caaggagtac aagtgcaagg tctccaacaa aggcctccca 1560gcccccatcg agaaaaccat ctccaaaacc aaaggtggga cccgcggggt atgagggcca 1620catggacaga ggccggctcg gcccaccctc tgccctggga gtgaccgctg tgccaacctc 1680tgtccctaca gggcagcccc gagaaccaca ggtgtacacc ctgcccccat cccgggagga 1740gatgaccaag aaccaggtca gcctgacctg cctggtcaaa ggcttctacc ccagcgacat 1800cgccgtggag tgggagagca atgggcagcc ggagaacaac tacaagacca cacctcccat 1860gctggactcc gacggctcct tcttcctcta cagcaagctc accgtggaca agagcaggtg 1920gcagcagggg aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac 1980gcagaagagc ctctccctgt ctccgggtaa a 2011812011DNAHomo sapiens 81tttacccgga gacagggaga ggctcttctg cgtgtagtgg ttgtgcagag cctcatgcat 60cacggagcat gagaagacgt tcccctgctg ccacctgctc ttgtccacgg tgagcttgct 120gtagaggaag aaggagccgt cggagtccag catgggaggt gtggtcttgt agttgttctc 180cggctgccca ttgctctccc actccacggc gatgtcgctg gggtagaagc ctttgaccag 240gcaggtcagg ctgacctggt tcttggtcat ctcctcccgg gatgggggca gggtgtacac 300ctgtggttct cggggctgcc ctgtagggac agaggttggc acagcggtca ctcccagggc 360agagggtggg ccgagccggc ctctgtccat gtggccctca taccccgcgg gtcccacctt 420tggttttgga gatggttttc tcgatggggg ctgggaggcc tttgttggag accttgcact 480tgtactcctt gccgttcagc cagtcctggt gcacaacggt gaggacgctg accacacgga 540acgtgctgtt gaactgctcc tcccgtggct ttgtcttggc attatgcacc tccacgccgt 600ccacgtacca gttgaactgg acctcggggt cttcgtggct cacgtccacc accacgcacg 660tgacctcagg ggtccgggag atcatgaggg tgtccttggg ttttgggggg aagaggaaga 720ctgacggtcc tgccacaggt ggtgctgagg aagagatgga ggtggacgtg tcagcaccca 780gctggggcct gtccctggat gcaggctact ctagggcacc tgtcccgcct tgagctggag 840ggcgaggcct gggctggctt acctgggcac ggtgggcact cgacacaaca tttgcgctct 900gcagagagaa gattgggagt tactcggatc tgggaggaga gaaggtgtcc gagctgaggg 960agtggacagt ttggcctttg gggtcggctt aggtcagggg cagggtcctc ccggatatgg 1020cttttggcag gtctgagcca agcacctgcc cctgtgtgtg aagggcctgg ggtaggggca 1080cccagcctgt gcctgcctgg agcctggtgg aaaaagccag aagaccctct ccctgagcat 1140gagtggggcg ggcagaggcc tccgggtgag gagacagatg gggcctgcct tgctgccctg 1200ggctggggct gcacagccgg ggtgcgtcca ggcaggaggg ctgagcctgg cttccagcag 1260acaccctccc tccctgagct ggcctctcac caactgtctt gtccaccttg gtgttgctgg 1320gcttgtgatc tacgttgcag gtgtaggtct gggtgccgaa gttgctggag ggcacggtca 1380ccacgctgct gagggagtag agtcctgagg actgtaggac agctgggaag gtgtgcacgc 1440cgctggtcag agcgcctgag ttccacgaca ccgtcaccgg ttcggggaag tagtccttga 1500ccaggcagcc cagggcggct gtgctctcgg aggtgcttct agagcagggc gccaggggga 1560agaccgatgg gcccttggtg gaggctgagg agacggtgac cgtggtccct tggccccaga 1620cgtccatacc gtagtcgtag acgtacctcc caccgtgatc tctcgcacag taatacacag 1680ccgtgtcctc aactctcagg ctgttcattt gcagatacag cgtgttcttg gaattgtctc 1740tggagatggt gaatcggccc ttcacggagt ctgcatagta tttattactt ccatcatatg 1800atataactgc cacccactcc agccccttgc ctggagcctg gcggacccag tgcatgccat 1860agctactgaa ggtgaatcca gaggctacac aggagagtct cagggacctc ccaggctgga 1920ccacgcctcc cccagactcc accagctgca cctgacactg gacacctctt aaaagagcaa 1980cgaggaaaac ccagcggagc cccaattcca t 201182252PRTHomo sapiens 82Met Glu Leu Gly Leu Arg Trp Val Phe Leu Val Ala Leu Ala Leu Leu1 5 10 15Arg Gly Val Gln Cys Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val 20 25 30Val Gln Pro Gly Arg Ser Leu Arg Leu Ser Cys Val Ala Leu Ala Ser 35 40 45Gly Phe Thr Phe Ser Ser Tyr Gly Met His Trp Val Ala Arg Gly Gln 50 55 60Ala Pro Gly Lys Gly Leu Glu Trp Val Ala Leu Ala Val Ile Ser Tyr65 70 75 80Asp Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr 85 90 95Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser 100 105 110Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp His Gly 115 120 125Gly Arg Tyr Val Tyr Asp Tyr Gly Met Asp Val Trp Gly Gln Gly Thr 130 135 140Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro145 150 155 160Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly 165 170 175Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 180 185 190Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 195 200 205Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 210 215 220Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Ala Ser Pro His Lys225 230 235 240Pro Ser Asn Thr Lys Val Ala Ser Pro Lys Thr Val 245 25083752DNAHomo sapiens 83atgagggtcc ccgctcagct cctggggctc ctgctgctct ggttcccagg ttccagatgc 60gacatccaga tgacccaatc tccatcttcc gtgtctgcat ctataggaga cagagtctcc 120atcacttgtc gggcgagtca gggtattagc agctggttag cctggtatca gcagaaacca 180gggaaagccc ctacgctcct tatctatgct gcatccactt tgcaacgtgg ggtcccatca 240aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag cctgcagcct 300gaagattttg caacttactt ttgtcaacag gctaacagtt tcccattcac tttcggccct 360gggaccaaag tggatatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcca 420tctgatgagc agttgaaatc tggaactgct agcgttgtgt gcctgctgaa taacttctat 480cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 540gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 600ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 660ctgagctcgc ccgtcacaaa gagcttcaac aggggaagtg ggtagtcccg gactcgagcg 720ggcagtgttt ctcgaagttg tcccctgagt gt 75284752DNAHomo sapiens 84acactcaggg gacaacttcg agaaacactg cccgctcgag tccgggacta cccacttccc 60ctgttgaagc tctttgtgac gggcgagctc aggccctgat gggtgacttc gcaggcgtag 120actttgtgtt tctcgtagtc tgctttgctc agcgtcaggg tgctgctgag gctgtaggtg 180ctgtccttgc tgtcctgctc tgtgacactc tcctgggagt tacccgattg gagggcgtta 240tccaccttcc actgtacttt ggcctctctg ggatagaagt tattcagcag gcacacaacg 300ctagcagttc cagatttcaa ctgctcatca gatggcggga agatgaagac agatggtgca 360gccacagttc gtttgatatc cactttggtc ccagggccga aagtgaatgg gaaactgtta 420gcctgttgac aaaagtaagt tgcaaaatct tcaggctgca ggctgctgat ggtgagagtg 480aaatctgtcc cagatccact gccgctgaac cttgatggga ccccacgttg caaagtggat 540gcagcataga taaggagcgt aggggctttc cctggtttct gctgatacca ggctaaccag 600ctgctaatac cctgactcgc ccgacaagtg atggagactc tgtctcctat agatgcagac 660acggaagatg gagattgggt catctggatg tcgcatctgg aacctgggaa ccagagcagc 720aggagcccca ggagctgagc ggggaccctc at 75285234PRTHomo sapiens 85Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro1 5 10 15Gly Ser Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser 20 25 30Ala Ser Ile Gly Asp Arg Val Ser Ile Thr Cys Arg Ala Ser Gln Gly 35 40 45Ile Ser Ser Trp Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 50 55 60Thr Leu Leu Ile Tyr Ala Ala Ser Thr Leu Gln Arg Gly Val Pro Ser65 70 75 80Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 85 90 95Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Ala Asn 100 105 110Ser Phe Pro Phe Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Arg 115 120 125Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr145 150 155 160Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230861990DNAHomo sapiens 86atgaagcatc tgtggttctt ccttctcctg gtggcagctc ccagatgggt cctgtcccag 60gtgcagctgc aggagtcggg cccaggactg gtgaagcctt cggagaccct gtccctcacc 120tgcactgtct ctggtgcctc catcagtaat tactactgga gctggatccg gcagccccca 180gggaagggac tggagtggat tgggtatgtc tcttacagtg ggagtacgta ctacaacccc 240tccctcaagg gtcgagtcac catgtcagta gacacgtcca agaaccagtt ctccctgaag 300ctgagctctg tgaccgctgc ggacacggcc gtgtattact gtgcgagaga aaaactgggg 360attggagact actggggcca gggaaccctg gtcaccgtct cctcagcctc caccaagggc 420ccatcggtct tccccctggc gccctgctct agaagcacct ccgagagcac agccgccctg 480ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgct 540ctgaccagcg gcgtgcacac cttcccagct gtcctacagt cctcaggact ctactccctc 600agcagcgtgg tgaccgtgcc ctccagcaac ttcggcaccc agacctacac ctgcaacgta 660gatcacaagc ccagcaacac caaggtggac aagacagttg gtgagaggcc agctcaggga 720gggagggtgt ctgctggaag ccaggctcag ccctcctgcc tggacgcacc ccggctgtgc 780agccccagcc cagggcagca aggcaggccc catctgtctc ctcacccgga ggcctctgcc 840cgccccactc atgctcaggg agagggtctt ctggcttttt ccaccaggct ccaggcaggc 900acaggctggg tgcccctacc ccaggccctt cacacacagg ggcaggtgct tggctcagac 960ctgccaaaag ccatatccgg gaggaccctg cccctgacct aagccgaccc caaaggccaa 1020actgtccact ccctcagctc ggacaccttc tctcctccca gatccgagta actcccaatc 1080ttctctctgc agagcgcaaa tgttgtgtcg agtgcccacc gtgcccaggt aagccagccc 1140aggcctcgcc ctccagctca aggcgggaca ggtgccctag agtagcctgc atccagggac 1200aggccccagc tgggtgctga cacgtccacc tccatctctt cctcagcacc acctgtggca 1260ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc 1320cctgaggtca cgtgcgtggt ggtggacgtg agccacgaag accccgaggt ccagttcaac 1380tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccacggga ggagcagttc 1440aacagcacgt tccgtgtggt cagcgtcctc accgttgtgc accaggactg gctgaacggc 1500aaggagtaca agtgcaaggt ctccaacaaa ggcctcccag cccccatcga gaaaaccatc 1560tccaaaacca aaggtgggac ccgcggggta tgagggccac atggacagag gccggctcgg 1620cccaccctct gccctgggag tgaccgctgt gccaacctct gtccctacag ggcagccccg 1680agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag 1740cctgacctgc ctggtcaaag gcttctaccc cagcgacatc gccgtggagt gggagagcaa 1800tgggcagccg gagaacaact acaagaccac acctcccatg ctggactccg acggctcctt 1860cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc 1920atgctccgtg atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc 1980tccgggtaaa 1990871990DNAHomo sapiens 87tttacccgga gacagggaga ggctcttctg cgtgtagtgg ttgtgcagag cctcatgcat 60cacggagcat gagaagacgt tcccctgctg ccacctgctc ttgtccacgg tgagcttgct 120gtagaggaag aaggagccgt cggagtccag catgggaggt gtggtcttgt agttgttctc 180cggctgccca ttgctctccc actccacggc gatgtcgctg gggtagaagc ctttgaccag 240gcaggtcagg ctgacctggt tcttggtcat ctcctcccgg gatgggggca gggtgtacac 300ctgtggttct cggggctgcc ctgtagggac agaggttggc acagcggtca ctcccagggc 360agagggtggg ccgagccggc ctctgtccat gtggccctca taccccgcgg gtcccacctt 420tggttttgga gatggttttc tcgatggggg ctgggaggcc tttgttggag accttgcact 480tgtactcctt gccgttcagc cagtcctggt gcacaacggt gaggacgctg accacacgga 540acgtgctgtt gaactgctcc tcccgtggct ttgtcttggc attatgcacc tccacgccgt 600ccacgtacca gttgaactgg acctcggggt cttcgtggct cacgtccacc accacgcacg 660tgacctcagg ggtccgggag atcatgaggg tgtccttggg ttttgggggg aagaggaaga 720ctgacggtcc tgccacaggt ggtgctgagg aagagatgga ggtggacgtg tcagcaccca 780gctggggcct gtccctggat gcaggctact ctagggcacc tgtcccgcct tgagctggag 840ggcgaggcct gggctggctt acctgggcac ggtgggcact cgacacaaca tttgcgctct 900gcagagagaa gattgggagt tactcggatc tgggaggaga gaaggtgtcc gagctgaggg 960agtggacagt ttggcctttg gggtcggctt aggtcagggg cagggtcctc ccggatatgg 1020cttttggcag gtctgagcca agcacctgcc cctgtgtgtg aagggcctgg ggtaggggca 1080cccagcctgt gcctgcctgg agcctggtgg aaaaagccag aagaccctct ccctgagcat 1140gagtggggcg ggcagaggcc tccgggtgag gagacagatg gggcctgcct tgctgccctg 1200ggctggggct gcacagccgg ggtgcgtcca ggcaggaggg ctgagcctgg cttccagcag 1260acaccctccc tccctgagct ggcctctcac caactgtctt gtccaccttg gtgttgctgg 1320gcttgtgatc tacgttgcag gtgtaggtct gggtgccgaa gttgctggag ggcacggtca 1380ccacgctgct gagggagtag agtcctgagg actgtaggac agctgggaag gtgtgcacgc 1440cgctggtcag agcgcctgag ttccacgaca ccgtcaccgg ttcggggaag tagtccttga 1500ccaggcagcc cagggcggct gtgctctcgg aggtgcttct agagcagggc gccaggggga 1560agaccgatgg gcccttggtg gaggctgagg agacggtgac cagggttccc tggccccagt 1620agtctccaat ccccagtttt tctctcgcac agtaatacac ggccgtgtcc gcagcggtca 1680cagagctcag cttcagggag aactggttct tggacgtgtc tactgacatg gtgactcgac 1740ccttgaggga ggggttgtag tacgtactcc cactgtaaga gacataccca atccactcca 1800gtcccttccc tgggggctgc cggatccagc tccagtagta attactgatg gaggcaccag 1860agacagtgca ggtgagggac agggtctccg aaggcttcac cagtcctggg cccgactcct 1920gcagctgcac ctgggacagg acccatctgg gagctgccac caggagaagg aagaaccaca 1980gatgcttcat 199088241PRTHomo sapiens 88Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Leu Ala Ala Pro1 5 10 15Arg Trp Val Leu Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu 20 25 30Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ala 35 40 45Ser Ile Ser Asn Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys 50 55 60Gly Leu Glu Trp Ile Gly Tyr Val Ser Tyr Ser Gly Ser Thr Tyr Tyr65 70 75 80Asn Pro Ser Leu Lys Gly Arg Val Thr Met Ser Val Ala Ser Pro Thr 85 90 95Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp 100 105 110Thr Ala Val Tyr Tyr Cys Ala Arg Glu Lys Leu Gly Ile Gly Asp Tyr 115 120 125Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 130 135 140Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser145 150 155 160Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 165 170 175Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 180 185 190Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 195 200 205Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 210 215 220Ala Ser Pro His Lys Pro Ser Asn Thr Lys Val Ala Ser Pro Lys Thr225 230 235 240Val89702DNAHomo sapiens 89atgaggctcc ccgctcagct cctggggctc ctgctgctct ggttcccagg tgccaggtgt 60gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 120atcacttgcc gggcaagtca gggcattaaa aatgatttag gctggtatca gcagaaacca 180gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 240aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 300gaagattttg caacttatta ctgtctacag cataatagtt atccgtgcag ttttggccag 360gggaccaagc tggagatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcca 420tctgatgagc agttgaaatc tggaactgct agcgttgtgt gcctgctgaa taacttctat 480cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 540gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 600ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 660ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 70290702DNAHomo sapiens 90acactctccc ctgttgaagc tctttgtgac gggcgagctc aggccctgat gggtgacttc 60gcaggcgtag actttgtgtt tctcgtagtc tgctttgctc agcgtcaggg tgctgctgag 120gctgtaggtg ctgtccttgc tgtcctgctc tgtgacactc tcctgggagt tacccgattg 180gagggcgtta tccaccttcc actgtacttt ggcctctctg ggatagaagt tattcagcag 240gcacacaacg ctagcagttc cagatttcaa ctgctcatca gatggcggga agatgaagac 300agatggtgca gccacagttc gtttgatctc cagcttggtc ccctggccaa aactgcacgg 360ataactatta tgctgtagac agtaataagt tgcaaaatct tcaggctgca ggctgctgat 420tgtgagagtg aattctgtcc cagatccact gccgctgaac cttgatggga ccccactttg 480caaactggat gcagcataga tcaggcgctt aggggctttc cctggtttct gctgatacca 540gcctaaatca tttttaatgc cctgacttgc ccggcaagtg atggtgactc tgtctcctac 600agatgcagac agggaggatg gagactgggt catctggatg tcacacctgg cacctgggaa 660ccagagcagc aggagcccca

ggagctgagc ggggagcctc at 70291234PRTHomo sapiens 91Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro1 5 10 15Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 20 25 30Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly 35 40 45Ile Lys Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 50 55 60Lys Arg Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser65 70 75 80Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser 85 90 95Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn 100 105 110Ser Tyr Pro Cys Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr145 150 155 160Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230921996DNAHomo sapiens 92atgaaacatc tgtggttctt cctcctgctg gtggcagctc ccagatgggt cctgtcccag 60gtgcagctgc aggagtcggg cccaggactg gtgaagcctt cacagaccct gtccctcacc 120tgcactgtct ctggtgcctc catcagcagt ggtgcttact actggagttg gatccgccag 180cacccaggga agggcctgga gtggattggg tacatctata agagtgagac ctcctactac 240aacccgtccc tcaagagtcg acttacccta tcagtagaca cgtctaagaa ccagttctcc 300ctgaacctga tctctgtgac tgccgcggac acggccgtgt attattgtgc gagagataaa 360ctggggatcg cggactactg gggccaggga accctggtca ccgtctcctc agcctccacc 420aagggcccat cggtcttccc cctggcgccc tgctctagaa gcacctccga gagcacagcc 480gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 540ggcgctctga ccagcggcgt gcacaccttc ccagctgtcc tacagtcctc aggactctac 600tccctcagca gcgtggtgac cgtgccctcc agcaacttcg gcacccagac ctacacctgc 660aacgtagatc acaagcccag caacaccaag gtggacaaga cagttggtga gaggccagct 720cagggaggga gggtgtctgc tggaagccag gctcagccct cctgcctgga cgcaccccgg 780ctgtgcagcc ccagcccagg gcagcaaggc aggccccatc tgtctcctca cccggaggcc 840tctgcccgcc ccactcatgc tcagggagag ggtcttctgg ctttttccac caggctccag 900gcaggcacag gctgggtgcc cctaccccag gcccttcaca cacaggggca ggtgcttggc 960tcagacctgc caaaagccat atccgggagg accctgcccc tgacctaagc cgaccccaaa 1020ggccaaactg tccactccct cagctcggac accttctctc ctcccagatc cgagtaactc 1080ccaatcttct ctctgcagag cgcaaatgtt gtgtcgagtg cccaccgtgc ccaggtaagc 1140cagcccaggc ctcgccctcc agctcaaggc gggacaggtg ccctagagta gcctgcatcc 1200agggacaggc cccagctggg tgctgacacg tccacctcca tctcttcctc agcaccacct 1260gtggcaggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct catgatctcc 1320cggacccctg aggtcacgtg cgtggtggtg gacgtgagcc acgaagaccc cgaggtccag 1380ttcaactggt acgtggacgg cgtggaggtg cataatgcca agacaaagcc acgggaggag 1440cagttcaaca gcacgttccg tgtggtcagc gtcctcaccg ttgtgcacca ggactggctg 1500aacggcaagg agtacaagtg caaggtctcc aacaaaggcc tcccagcccc catcgagaaa 1560accatctcca aaaccaaagg tgggacccgc ggggtatgag ggccacatgg acagaggccg 1620gctcggccca ccctctgccc tgggagtgac cgctgtgcca acctctgtcc ctacagggca 1680gccccgagaa ccacaggtgt acaccctgcc cccatcccgg gaggagatga ccaagaacca 1740ggtcagcctg acctgcctgg tcaaaggctt ctaccccagc gacatcgccg tggagtggga 1800gagcaatggg cagccggaga acaactacaa gaccacacct cccatgctgg actccgacgg 1860ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt 1920cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc 1980cctgtctccg ggtaaa 1996931996DNAHomo sapiens 93tttacccgga gacagggaga ggctcttctg cgtgtagtgg ttgtgcagag cctcatgcat 60cacggagcat gagaagacgt tcccctgctg ccacctgctc ttgtccacgg tgagcttgct 120gtagaggaag aaggagccgt cggagtccag catgggaggt gtggtcttgt agttgttctc 180cggctgccca ttgctctccc actccacggc gatgtcgctg gggtagaagc ctttgaccag 240gcaggtcagg ctgacctggt tcttggtcat ctcctcccgg gatgggggca gggtgtacac 300ctgtggttct cggggctgcc ctgtagggac agaggttggc acagcggtca ctcccagggc 360agagggtggg ccgagccggc ctctgtccat gtggccctca taccccgcgg gtcccacctt 420tggttttgga gatggttttc tcgatggggg ctgggaggcc tttgttggag accttgcact 480tgtactcctt gccgttcagc cagtcctggt gcacaacggt gaggacgctg accacacgga 540acgtgctgtt gaactgctcc tcccgtggct ttgtcttggc attatgcacc tccacgccgt 600ccacgtacca gttgaactgg acctcggggt cttcgtggct cacgtccacc accacgcacg 660tgacctcagg ggtccgggag atcatgaggg tgtccttggg ttttgggggg aagaggaaga 720ctgacggtcc tgccacaggt ggtgctgagg aagagatgga ggtggacgtg tcagcaccca 780gctggggcct gtccctggat gcaggctact ctagggcacc tgtcccgcct tgagctggag 840ggcgaggcct gggctggctt acctgggcac ggtgggcact cgacacaaca tttgcgctct 900gcagagagaa gattgggagt tactcggatc tgggaggaga gaaggtgtcc gagctgaggg 960agtggacagt ttggcctttg gggtcggctt aggtcagggg cagggtcctc ccggatatgg 1020cttttggcag gtctgagcca agcacctgcc cctgtgtgtg aagggcctgg ggtaggggca 1080cccagcctgt gcctgcctgg agcctggtgg aaaaagccag aagaccctct ccctgagcat 1140gagtggggcg ggcagaggcc tccgggtgag gagacagatg gggcctgcct tgctgccctg 1200ggctggggct gcacagccgg ggtgcgtcca ggcaggaggg ctgagcctgg cttccagcag 1260acaccctccc tccctgagct ggcctctcac caactgtctt gtccaccttg gtgttgctgg 1320gcttgtgatc tacgttgcag gtgtaggtct gggtgccgaa gttgctggag ggcacggtca 1380ccacgctgct gagggagtag agtcctgagg actgtaggac agctgggaag gtgtgcacgc 1440cgctggtcag agcgcctgag ttccacgaca ccgtcaccgg ttcggggaag tagtccttga 1500ccaggcagcc cagggcggct gtgctctcgg aggtgcttct agagcagggc gccaggggga 1560agaccgatgg gcccttggtg gaggctgagg agacggtgac cagggttccc tggccccagt 1620agtccgcgat ccccagttta tctctcgcac aataatacac ggccgtgtcc gcggcagtca 1680cagagatcag gttcagggag aactggttct tagacgtgtc tactgatagg gtaagtcgac 1740tcttgaggga cgggttgtag taggaggtct cactcttata gatgtaccca atccactcca 1800ggcccttccc tgggtgctgg cggatccaac tccagtagta agcaccactg ctgatggagg 1860caccagagac agtgcaggtg agggacaggg tctgtgaagg cttcaccagt cctgggcccg 1920actcctgcag ctgcacctgg gacaggaccc atctgggagc tgccaccagc aggaggaaga 1980accacagatg tttcat 199694243PRTHomo sapiens 94Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Leu Ala Ala Pro1 5 10 15Arg Trp Val Leu Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu 20 25 30Val Lys Pro Ser Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Ala 35 40 45Ser Ile Ser Ser Gly Ala Tyr Tyr Trp Ser Trp Ile Arg Gln His Pro 50 55 60Gly Lys Gly Leu Glu Trp Ile Gly Tyr Ile Tyr Lys Ser Glu Thr Ser65 70 75 80Tyr Tyr Asn Pro Ser Leu Lys Ser Arg Leu Thr Leu Ser Val Ala Ser 85 90 95Pro Thr Ser Lys Asn Gln Phe Ser Leu Asn Leu Ile Ser Val Thr Ala 100 105 110Ala Asp Thr Ala Val Tyr Tyr Cys Ala Arg Asp Lys Leu Gly Ile Ala 115 120 125Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 130 135 140Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser145 150 155 160Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200 205Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys 210 215 220Asn Val Ala Ser Pro His Lys Pro Ser Asn Thr Lys Val Ala Ser Pro225 230 235 240Lys Thr Val95702DNAHomo sapiens 95atgagggtcc ccgctcagct cctggggctc ctgctgctct ggttcccagg cgccaggtgt 60gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 120atcacttgcc gggcaagtca ggacattaga aatgatttag gctggtatca gcagaaacca 180gggaaagccc ctaagcgcct gatctatgct gcatccaatt tgcaaagtgg ggtcccatca 240aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 300gaagattttg caacttatta ctgtctacag cataatagct accctcccac tttcggcgga 360gggaccaagg tggaaatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcca 420tctgatgagc agttgaaatc tggaactgct agcgttgtgt gcctgctgaa taacttctat 480cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 540gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 600ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 660ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 70296702DNAHomo sapiens 96acactctccc ctgttgaagc tctttgtgac gggcgagctc aggccctgat gggtgacttc 60gcaggcgtag actttgtgtt tctcgtagtc tgctttgctc agcgtcaggg tgctgctgag 120gctgtaggtg ctgtccttgc tgtcctgctc tgtgacactc tcctgggagt tacccgattg 180gagggcgtta tccaccttcc actgtacttt ggcctctctg ggatagaagt tattcagcag 240gcacacaacg ctagcagttc cagatttcaa ctgctcatca gatggcggga agatgaagac 300agatggtgca gccacagttc gtttgatttc caccttggtc cctccgccga aagtgggagg 360gtagctatta tgctgtagac agtaataagt tgcaaaatct tcaggctgca ggctgctgat 420tgtgagagtg aattctgtcc cagatccact gccgctgaac cttgatggga ccccactttg 480caaattggat gcagcataga tcaggcgctt aggggctttc cctggtttct gctgatacca 540gcctaaatca tttctaatgt cctgacttgc ccggcaagtg atggtgactc tgtctcctac 600agatgcagac agggaggatg gagactgggt catctggatg tcacacctgg cgcctgggaa 660ccagagcagc aggagcccca ggagctgagc ggggaccctc at 70297234PRTHomo sapiens 97Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro1 5 10 15Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 20 25 30Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Asp 35 40 45Ile Arg Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 50 55 60Lys Arg Leu Ile Tyr Ala Ala Ser Asn Leu Gln Ser Gly Val Pro Ser65 70 75 80Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser 85 90 95Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn 100 105 110Ser Tyr Pro Pro Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 115 120 125Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr145 150 155 160Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230981990DNAHomo sapiens 98atgaaacacc tgtggttctt ccttctcctg gtggcagctc ccagatgggt cctgtcccag 60gtgcagctgc aggagtcggg cccaggactg gtgaagcctt cggagaccct gtccctcacc 120tgcactgtct ctggtgtctc catcagtaat tactactgga gctggatccg gcagtcccca 180gggaagggac tggagtggat tggatatatc tattacagtg ggagtcccta ttacaacccc 240tccctcaaga gtcgagtcac tatatctgca gacacgtcca agaaccaatt ctccctgaag 300ctgagctctg tgaccgctgc ggacacggcc atttattact gtgcgagaga aaaactgggg 360attggagact actggggcca gggaaccctg gtcaccgtct cctcagcctc caccaagggc 420ccatcggtct tccccctggc gccctgctct agaagcacct ccgagagcac agccgccctg 480ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgct 540ctgaccagcg gcgtgcacac cttcccagct gtcctacagt cctcaggact ctactccctc 600agcagcgtgg tgaccgtgcc ctccagcaac ttcggcaccc agacctacac ctgcaacgta 660gatcacaagc ccagcaacac caaggtggac aagacagttg gtgagaggcc agctcaggga 720gggagggtgt ctgctggaag ccaggctcag ccctcctgcc tggacgcacc ccggctgtgc 780agccccagcc cagggcagca aggcaggccc catctgtctc ctcacccgga ggcctctgcc 840cgccccactc atgctcaggg agagggtctt ctggcttttt ccaccaggct ccaggcaggc 900acaggctggg tgcccctacc ccaggccctt cacacacagg ggcaggtgct tggctcagac 960ctgccaaaag ccatatccgg gaggaccctg cccctgacct aagccgaccc caaaggccaa 1020actgtccact ccctcagctc ggacaccttc tctcctccca gatccgagta actcccaatc 1080ttctctctgc agagcgcaaa tgttgtgtcg agtgcccacc gtgcccaggt aagccagccc 1140aggcctcgcc ctccagctca aggcgggaca ggtgccctag agtagcctgc atccagggac 1200aggccccagc tgggtgctga cacgtccacc tccatctctt cctcagcacc acctgtggca 1260ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc 1320cctgaggtca cgtgcgtggt ggtggacgtg agccacgaag accccgaggt ccagttcaac 1380tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccacggga ggagcagttc 1440aacagcacgt tccgtgtggt cagcgtcctc accgttgtgc accaggactg gctgaacggc 1500aaggagtaca agtgcaaggt ctccaacaaa ggcctcccag cccccatcga gaaaaccatc 1560tccaaaacca aaggtgggac ccgcggggta tgagggccac atggacagag gccggctcgg 1620cccaccctct gccctgggag tgaccgctgt gccaacctct gtccctacag ggcagccccg 1680agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag 1740cctgacctgc ctggtcaaag gcttctaccc cagcgacatc gccgtggagt gggagagcaa 1800tgggcagccg gagaacaact acaagaccac acctcccatg ctggactccg acggctcctt 1860cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc 1920atgctccgtg atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc 1980tccgggtaaa 1990991990DNAHomo sapiens 99tttacccgga gacagggaga ggctcttctg cgtgtagtgg ttgtgcagag cctcatgcat 60cacggagcat gagaagacgt tcccctgctg ccacctgctc ttgtccacgg tgagcttgct 120gtagaggaag aaggagccgt cggagtccag catgggaggt gtggtcttgt agttgttctc 180cggctgccca ttgctctccc actccacggc gatgtcgctg gggtagaagc ctttgaccag 240gcaggtcagg ctgacctggt tcttggtcat ctcctcccgg gatgggggca gggtgtacac 300ctgtggttct cggggctgcc ctgtagggac agaggttggc acagcggtca ctcccagggc 360agagggtggg ccgagccggc ctctgtccat gtggccctca taccccgcgg gtcccacctt 420tggttttgga gatggttttc tcgatggggg ctgggaggcc tttgttggag accttgcact 480tgtactcctt gccgttcagc cagtcctggt gcacaacggt gaggacgctg accacacgga 540acgtgctgtt gaactgctcc tcccgtggct ttgtcttggc attatgcacc tccacgccgt 600ccacgtacca gttgaactgg acctcggggt cttcgtggct cacgtccacc accacgcacg 660tgacctcagg ggtccgggag atcatgaggg tgtccttggg ttttgggggg aagaggaaga 720ctgacggtcc tgccacaggt ggtgctgagg aagagatgga ggtggacgtg tcagcaccca 780gctggggcct gtccctggat gcaggctact ctagggcacc tgtcccgcct tgagctggag 840ggcgaggcct gggctggctt acctgggcac ggtgggcact cgacacaaca tttgcgctct 900gcagagagaa gattgggagt tactcggatc tgggaggaga gaaggtgtcc gagctgaggg 960agtggacagt ttggcctttg gggtcggctt aggtcagggg cagggtcctc ccggatatgg 1020cttttggcag gtctgagcca agcacctgcc cctgtgtgtg aagggcctgg ggtaggggca 1080cccagcctgt gcctgcctgg agcctggtgg aaaaagccag aagaccctct ccctgagcat 1140gagtggggcg ggcagaggcc tccgggtgag gagacagatg gggcctgcct tgctgccctg 1200ggctggggct gcacagccgg ggtgcgtcca ggcaggaggg ctgagcctgg cttccagcag 1260acaccctccc tccctgagct ggcctctcac caactgtctt gtccaccttg gtgttgctgg 1320gcttgtgatc tacgttgcag gtgtaggtct gggtgccgaa gttgctggag ggcacggtca 1380ccacgctgct gagggagtag agtcctgagg actgtaggac agctgggaag gtgtgcacgc 1440cgctggtcag agcgcctgag ttccacgaca ccgtcaccgg ttcggggaag tagtccttga 1500ccaggcagcc cagggcggct gtgctctcgg aggtgcttct agagcagggc gccaggggga 1560agaccgatgg gcccttggtg gaggctgagg agacggtgac cagggttccc tggccccagt 1620agtctccaat ccccagtttt tctctcgcac agtaataaat ggccgtgtcc gcagcggtca 1680cagagctcag cttcagggag aattggttct tggacgtgtc tgcagatata gtgactcgac 1740tcttgaggga ggggttgtaa tagggactcc cactgtaata gatatatcca atccactcca 1800gtcccttccc tggggactgc cggatccagc tccagtagta attactgatg gagacaccag 1860agacagtgca ggtgagggac agggtctccg aaggcttcac cagtcctggg cccgactcct 1920gcagctgcac ctgggacagg acccatctgg gagctgccac caggagaagg aagaaccaca 1980ggtgtttcat 1990100239PRTHomo sapiens 100Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Leu Ala Ala Pro1 5 10 15Arg Trp Val Leu Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu 20 25 30Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Val 35 40 45Ser Ile Ser Asn Tyr Tyr Trp Ser Trp Ile Arg Gln Ser Pro Gly Lys 50 55 60Gly Leu Glu Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser Pro Tyr Tyr65 70 75 80Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Ala Asp Thr Ser Lys 85 90 95Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala 100 105 110Ile Tyr Tyr Cys Ala Arg Glu Lys Leu Gly Ile Gly Asp Tyr Trp Gly 115 120 125Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 130 135 140Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala145 150

155 160Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 165 170 175Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 180 185 190Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 195 200 205Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Ala Ser 210 215 220Pro His Lys Pro Ser Asn Thr Lys Val Ala Ser Pro Lys Thr Val225 230 235101702DNAHomo sapiens 101atgagggtcc ccgctcagct cctggggctc ctgctgctct ggttcccagg tgccaggtgt 60gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtcggaga cagagtcacc 120atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 180gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 240aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 300gaagattttg caacttatta ctgtctacag cataatagtt accctcccac tttcggccct 360gggaccaagg tggatatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcca 420tctgatgagc agttgaaatc tggaactgct agcgttgtgt gcctgctgaa taacttctat 480cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 540gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 600ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 660ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 702102702DNAHomo sapiens 102acactctccc ctgttgaagc tctttgtgac gggcgagctc aggccctgat gggtgacttc 60gcaggcgtag actttgtgtt tctcgtagtc tgctttgctc agcgtcaggg tgctgctgag 120gctgtaggtg ctgtccttgc tgtcctgctc tgtgacactc tcctgggagt tacccgattg 180gagggcgtta tccaccttcc actgtacttt ggcctctctg ggatagaagt tattcagcag 240gcacacaacg ctagcagttc cagatttcaa ctgctcatca gatggcggga agatgaagac 300agatggtgca gccacagttc gtttgatatc caccttggtc ccagggccga aagtgggagg 360gtaactatta tgctgtagac agtaataagt tgcaaaatct tcaggctgca ggctgctgat 420tgtgagagtg aattctgtcc cagatccact gccgctgaac cttgatggga ccccactttg 480caaactggat gcagcataga tcaggcgctt aggggctttc cctggtttct gctgatacca 540gcctaaatca tttctaatgc cctgacttgc ccggcaagtg atggtgactc tgtctccgac 600agatgcagac agggaggatg gagactgggt catctggatg tcacacctgg cacctgggaa 660ccagagcagc aggagcccca ggagctgagc ggggaccctc at 702103234PRTHomo sapiens 103Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro1 5 10 15Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 20 25 30Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly 35 40 45Ile Arg Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 50 55 60Lys Arg Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser65 70 75 80Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser 85 90 95Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn 100 105 110Ser Tyr Pro Pro Thr Phe Gly Pro Gly Thr Lys Val Asp Ile Lys Arg 115 120 125Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr145 150 155 160Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 2301041990DNAHomo sapiens 104atgaaacatc tgtggttctt ccttctcctg gtggcagctc ccagatgggt cctgtcccag 60gtgcagctgc aggagtcggg cccaggactg gtgaagcctt cggagaccct gtccctcacc 120tgcactgtct ctggtggctc catcagtcgt tactactgga gctggatccg gcagccccca 180gggaagggac tggagtggat tgggtatgtc tcttacagtg ggagcaccta ctacaacccc 240tccctcaaga gtcgagtcac catatcagta gacacgtcca agaaccagtt ctccctgaag 300ctgagctctg tgaccgctgc ggacacggcc gtgtattact gtgcgagaga taaactgggg 360attggagact actggggcca gggaaccctg gtcaccgtct cctcagcctc caccaagggc 420ccatcggtct tccccctggc gccctgctct agaagcacct ccgagagcac agccgccctg 480ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgct 540ctgaccagcg gcgtgcacac cttcccagct gtcctacagt cctcaggact ctactccctc 600agcagcgtgg tgaccgtgcc ctccagcaac ttcggcaccc agacctacac ctgcaacgta 660gatcacaagc ccagcaacac caaggtggac aagacagttg gtgagaggcc agctcaggga 720gggagggtgt ctgctggaag ccaggctcag ccctcctgcc tggacgcacc ccggctgtgc 780agccccagcc cagggcagca aggcaggccc catctgtctc ctcacccgga ggcctctgcc 840cgccccactc atgctcaggg agagggtctt ctggcttttt ccaccaggct ccaggcaggc 900acaggctggg tgcccctacc ccaggccctt cacacacagg ggcaggtgct tggctcagac 960ctgccaaaag ccatatccgg gaggaccctg cccctgacct aagccgaccc caaaggccaa 1020actgtccact ccctcagctc ggacaccttc tctcctccca gatccgagta actcccaatc 1080ttctctctgc agagcgcaaa tgttgtgtcg agtgcccacc gtgcccaggt aagccagccc 1140aggcctcgcc ctccagctca aggcgggaca ggtgccctag agtagcctgc atccagggac 1200aggccccagc tgggtgctga cacgtccacc tccatctctt cctcagcacc acctgtggca 1260ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc 1320cctgaggtca cgtgcgtggt ggtggacgtg agccacgaag accccgaggt ccagttcaac 1380tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccacggga ggagcagttc 1440aacagcacgt tccgtgtggt cagcgtcctc accgttgtgc accaggactg gctgaacggc 1500aaggagtaca agtgcaaggt ctccaacaaa ggcctcccag cccccatcga gaaaaccatc 1560tccaaaacca aaggtgggac ccgcggggta tgagggccac atggacagag gccggctcgg 1620cccaccctct gccctgggag tgaccgctgt gccaacctct gtccctacag ggcagccccg 1680agaaccacag gtgtacaccc tgcccccatc ccgggaggag atgaccaaga accaggtcag 1740cctgacctgc ctggtcaaag gcttctaccc cagcgacatc gccgtggagt gggagagcaa 1800tgggcagccg gagaacaact acaagaccac acctcccatg ctggactccg acggctcctt 1860cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc 1920atgctccgtg atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc 1980tccgggtaaa 19901051990DNAHomo sapiens 105tttacccgga gacagggaga ggctcttctg cgtgtagtgg ttgtgcagag cctcatgcat 60cacggagcat gagaagacgt tcccctgctg ccacctgctc ttgtccacgg tgagcttgct 120gtagaggaag aaggagccgt cggagtccag catgggaggt gtggtcttgt agttgttctc 180cggctgccca ttgctctccc actccacggc gatgtcgctg gggtagaagc ctttgaccag 240gcaggtcagg ctgacctggt tcttggtcat ctcctcccgg gatgggggca gggtgtacac 300ctgtggttct cggggctgcc ctgtagggac agaggttggc acagcggtca ctcccagggc 360agagggtggg ccgagccggc ctctgtccat gtggccctca taccccgcgg gtcccacctt 420tggttttgga gatggttttc tcgatggggg ctgggaggcc tttgttggag accttgcact 480tgtactcctt gccgttcagc cagtcctggt gcacaacggt gaggacgctg accacacgga 540acgtgctgtt gaactgctcc tcccgtggct ttgtcttggc attatgcacc tccacgccgt 600ccacgtacca gttgaactgg acctcggggt cttcgtggct cacgtccacc accacgcacg 660tgacctcagg ggtccgggag atcatgaggg tgtccttggg ttttgggggg aagaggaaga 720ctgacggtcc tgccacaggt ggtgctgagg aagagatgga ggtggacgtg tcagcaccca 780gctggggcct gtccctggat gcaggctact ctagggcacc tgtcccgcct tgagctggag 840ggcgaggcct gggctggctt acctgggcac ggtgggcact cgacacaaca tttgcgctct 900gcagagagaa gattgggagt tactcggatc tgggaggaga gaaggtgtcc gagctgaggg 960agtggacagt ttggcctttg gggtcggctt aggtcagggg cagggtcctc ccggatatgg 1020cttttggcag gtctgagcca agcacctgcc cctgtgtgtg aagggcctgg ggtaggggca 1080cccagcctgt gcctgcctgg agcctggtgg aaaaagccag aagaccctct ccctgagcat 1140gagtggggcg ggcagaggcc tccgggtgag gagacagatg gggcctgcct tgctgccctg 1200ggctggggct gcacagccgg ggtgcgtcca ggcaggaggg ctgagcctgg cttccagcag 1260acaccctccc tccctgagct ggcctctcac caactgtctt gtccaccttg gtgttgctgg 1320gcttgtgatc tacgttgcag gtgtaggtct gggtgccgaa gttgctggag ggcacggtca 1380ccacgctgct gagggagtag agtcctgagg actgtaggac agctgggaag gtgtgcacgc 1440cgctggtcag agcgcctgag ttccacgaca ccgtcaccgg ttcggggaag tagtccttga 1500ccaggcagcc cagggcggct gtgctctcgg aggtgcttct agagcagggc gccaggggga 1560agaccgatgg gcccttggtg gaggctgagg agacggtgac cagggttccc tggccccagt 1620agtctccaat ccccagttta tctctcgcac agtaatacac ggccgtgtcc gcagcggtca 1680cagagctcag cttcagggag aactggttct tggacgtgtc tactgatatg gtgactcgac 1740tcttgaggga ggggttgtag taggtgctcc cactgtaaga gacataccca atccactcca 1800gtcccttccc tgggggctgc cggatccagc tccagtagta acgactgatg gagccaccag 1860agacagtgca ggtgagggac agggtctccg aaggcttcac cagtcctggg cccgactcct 1920gcagctgcac ctgggacagg acccatctgg gagctgccac caggagaagg aagaaccaca 1980gatgtttcat 1990106241PRTHomo sapiens 106Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Leu Ala Ala Pro1 5 10 15Arg Trp Val Leu Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu 20 25 30Val Lys Pro Ser Glu Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly 35 40 45Ser Ile Ser Arg Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys 50 55 60Gly Leu Glu Trp Ile Gly Tyr Val Ser Tyr Ser Gly Ser Thr Tyr Tyr65 70 75 80Asn Pro Ser Leu Lys Ser Arg Val Thr Ile Ser Val Ala Ser Pro Thr 85 90 95Ser Lys Asn Gln Phe Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp 100 105 110Thr Ala Val Tyr Tyr Cys Ala Arg Asp Lys Leu Gly Ile Gly Asp Tyr 115 120 125Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly 130 135 140Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser145 150 155 160Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val 165 170 175Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 180 185 190Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 195 200 205Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 210 215 220Ala Ser Pro His Lys Pro Ser Asn Thr Lys Val Ala Ser Pro Lys Thr225 230 235 240Val107702DNAHomo sapiens 107atgaggctcc ctgctcagct cctggggctc ctgctgctct ggttcccagg tgccaggtgt 60gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 120atcacttgcc gggcaagtca gggcattaga aatgatttag gctggtatca gcagaaaccg 180gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 240aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 300gaagattttg caacttatta ctgtctacag cataatagtt acccgtgcag ttttggccag 360gggaccaagc tggagatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcca 420tctgatgagc agttgaaatc tggaactgct agcgttgtgt gcctgctgaa taacttctat 480cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 540gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 600ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 660ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 702108702DNAHomo sapiens 108acactctccc ctgttgaagc tctttgtgac gggcgagctc aggccctgat gggtgacttc 60gcaggcgtag actttgtgtt tctcgtagtc tgctttgctc agcgtcaggg tgctgctgag 120gctgtaggtg ctgtccttgc tgtcctgctc tgtgacactc tcctgggagt tacccgattg 180gagggcgtta tccaccttcc actgtacttt ggcctctctg ggatagaagt tattcagcag 240gcacacaacg ctagcagttc cagatttcaa ctgctcatca gatggcggga agatgaagac 300agatggtgca gccacagttc gtttgatctc cagcttggtc ccctggccaa aactgcacgg 360gtaactatta tgctgtagac agtaataagt tgcaaaatct tcaggctgca ggctgctgat 420tgtgagagtg aattctgtcc cagatccact gccgctgaac cttgatggga ccccactttg 480caaactggat gcagcataga tcaggcgctt aggggctttc cccggtttct gctgatacca 540gcctaaatca tttctaatgc cctgacttgc ccggcaagtg atggtgactc tgtctcctac 600agatgcagac agggaggatg gagactgggt catctggatg tcacacctgg cacctgggaa 660ccagagcagc aggagcccca ggagctgagc agggagcctc at 702109234PRTHomo sapiens 109Met Arg Leu Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro1 5 10 15Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 20 25 30Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly 35 40 45Ile Arg Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 50 55 60Lys Arg Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser65 70 75 80Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser 85 90 95Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn 100 105 110Ser Tyr Pro Cys Ser Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg 115 120 125Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr145 150 155 160Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 2301101996DNAHomo sapiens 110atgaagcatc tgtggttctt cctcctgctg gtggcagctc ccagatgggt cctgtcccag 60gtgcagctgc aggagtcggg cccaggactg gtgaagcctt tacagaccct gtccctcacc 120tgcactgtct ctggtggctc catcagcagt ggtgtttact actggagctg gatccgccag 180cacccaggga agggcctgga gtggattggg tacatctata acagtaagac ctcctattat 240aatccgtccc tcaagagtcg acttacccta tcagtagaca cgtctaagaa ccagttctcc 300ctgaacctga tctctgtgac tgccgcggac acggccgtgt attactgtgc gagagataaa 360ttggggatcg cggactactg gggccaggga accctggtca ccgtctcctc agcctccacc 420aagggcccat cggtcttccc cctggcgccc tgctctagaa gcacctccga gagcacagcc 480gccctgggct gcctggtcaa ggactacttc cccgaaccgg tgacggtgtc gtggaactca 540ggcgctctga ccagcggcgt gcacaccttc ccagctgtcc tacagtcctc aggactctac 600tccctcagca gcgtggtgac cgtgccctcc agcaacttcg gcacccagac ctacacctgc 660aacgtagatc acaagcccag caacaccaag gtggacaaga cagttggtga gaggccagct 720cagggaggga gggtgtctgc tggaagccag gctcagccct cctgcctgga cgcaccccgg 780ctgtgcagcc ccagcccagg gcagcaaggc aggccccatc tgtctcctca cccggaggcc 840tctgcccgcc ccactcatgc tcagggagag ggtcttctgg ctttttccac caggctccag 900gcaggcacag gctgggtgcc cctaccccag gcccttcaca cacaggggca ggtgcttggc 960tcagacctgc caaaagccat atccgggagg accctgcccc tgacctaagc cgaccccaaa 1020ggccaaactg tccactccct cagctcggac accttctctc ctcccagatc cgagtaactc 1080ccaatcttct ctctgcagag cgcaaatgtt gtgtcgagtg cccaccgtgc ccaggtaagc 1140cagcccaggc ctcgccctcc agctcaaggc gggacaggtg ccctagagta gcctgcatcc 1200agggacaggc cccagctggg tgctgacacg tccacctcca tctcttcctc agcaccacct 1260gtggcaggac cgtcagtctt cctcttcccc ccaaaaccca aggacaccct catgatctcc 1320cggacccctg aggtcacgtg cgtggtggtg gacgtgagcc acgaagaccc cgaggtccag 1380ttcaactggt acgtggacgg cgtggaggtg cataatgcca agacaaagcc acgggaggag 1440cagttcaaca gcacgttccg tgtggtcagc gtcctcaccg ttgtgcacca ggactggctg 1500aacggcaagg agtacaagtg caaggtctcc aacaaaggcc tcccagcccc catcgagaaa 1560accatctcca aaaccaaagg tgggacccgc ggggtatgag ggccacatgg acagaggccg 1620gctcggccca ccctctgccc tgggagtgac cgctgtgcca acctctgtcc ctacagggca 1680gccccgagaa ccacaggtgt acaccctgcc cccatcccgg gaggagatga ccaagaacca 1740ggtcagcctg acctgcctgg tcaaaggctt ctaccccagc gacatcgccg tggagtggga 1800gagcaatggg cagccggaga acaactacaa gaccacacct cccatgctgg actccgacgg 1860ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt 1920cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc 1980cctgtctccg ggtaaa 19961111996DNAHomo sapiens 111tttacccgga gacagggaga ggctcttctg cgtgtagtgg ttgtgcagag cctcatgcat 60cacggagcat gagaagacgt tcccctgctg ccacctgctc ttgtccacgg tgagcttgct 120gtagaggaag aaggagccgt cggagtccag catgggaggt gtggtcttgt agttgttctc 180cggctgccca ttgctctccc actccacggc gatgtcgctg gggtagaagc ctttgaccag 240gcaggtcagg ctgacctggt tcttggtcat ctcctcccgg gatgggggca gggtgtacac 300ctgtggttct cggggctgcc ctgtagggac agaggttggc acagcggtca ctcccagggc 360agagggtggg ccgagccggc ctctgtccat gtggccctca taccccgcgg gtcccacctt 420tggttttgga gatggttttc tcgatggggg ctgggaggcc tttgttggag accttgcact 480tgtactcctt gccgttcagc cagtcctggt gcacaacggt gaggacgctg accacacgga 540acgtgctgtt gaactgctcc tcccgtggct ttgtcttggc attatgcacc tccacgccgt 600ccacgtacca gttgaactgg acctcggggt cttcgtggct cacgtccacc accacgcacg 660tgacctcagg ggtccgggag atcatgaggg tgtccttggg ttttgggggg aagaggaaga 720ctgacggtcc tgccacaggt ggtgctgagg aagagatgga ggtggacgtg tcagcaccca 780gctggggcct gtccctggat gcaggctact ctagggcacc tgtcccgcct tgagctggag 840ggcgaggcct gggctggctt acctgggcac ggtgggcact cgacacaaca tttgcgctct 900gcagagagaa gattgggagt tactcggatc tgggaggaga gaaggtgtcc gagctgaggg 960agtggacagt ttggcctttg gggtcggctt aggtcagggg cagggtcctc ccggatatgg 1020cttttggcag gtctgagcca agcacctgcc cctgtgtgtg

aagggcctgg ggtaggggca 1080cccagcctgt gcctgcctgg agcctggtgg aaaaagccag aagaccctct ccctgagcat 1140gagtggggcg ggcagaggcc tccgggtgag gagacagatg gggcctgcct tgctgccctg 1200ggctggggct gcacagccgg ggtgcgtcca ggcaggaggg ctgagcctgg cttccagcag 1260acaccctccc tccctgagct ggcctctcac caactgtctt gtccaccttg gtgttgctgg 1320gcttgtgatc tacgttgcag gtgtaggtct gggtgccgaa gttgctggag ggcacggtca 1380ccacgctgct gagggagtag agtcctgagg actgtaggac agctgggaag gtgtgcacgc 1440cgctggtcag agcgcctgag ttccacgaca ccgtcaccgg ttcggggaag tagtccttga 1500ccaggcagcc cagggcggct gtgctctcgg aggtgcttct agagcagggc gccaggggga 1560agaccgatgg gcccttggtg gaggctgagg agacggtgac cagggttccc tggccccagt 1620agtccgcgat ccccaattta tctctcgcac agtaatacac ggccgtgtcc gcggcagtca 1680cagagatcag gttcagggag aactggttct tagacgtgtc tactgatagg gtaagtcgac 1740tcttgaggga cggattataa taggaggtct tactgttata gatgtaccca atccactcca 1800ggcccttccc tgggtgctgg cggatccagc tccagtagta aacaccactg ctgatggagc 1860caccagagac agtgcaggtg agggacaggg tctgtaaagg cttcaccagt cctgggcccg 1920actcctgcag ctgcacctgg gacaggaccc atctgggagc tgccaccagc aggaggaaga 1980accacagatg cttcat 1996112235PRThomo sapiens 112Met Lys His Leu Trp Phe Phe Leu Leu Leu Val Ala Ala Pro Arg Trp1 5 10 15Val Leu Ser Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys 20 25 30Pro Leu Gln Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile 35 40 45Ser Ser Gly Val Tyr Tyr Trp Ser Trp Ile Arg Gln His Pro Gly Lys 50 55 60Gly Leu Glu Trp Ile Gly Tyr Ile Tyr Asn Ser Lys Thr Ser Tyr Tyr65 70 75 80Asn Pro Ser Leu Lys Ser Arg Leu Thr Leu Ser Val Asp Thr Ser Lys 85 90 95Asn Gln Phe Ser Leu Asn Leu Ile Ser Val Thr Ala Ala Asp Thr Ala 100 105 110Val Tyr Tyr Cys Ala Arg Asp Lys Leu Gly Ile Ala Asp Tyr Trp Gly 115 120 125Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser 130 135 140Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala145 150 155 160Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val 165 170 175Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala 180 185 190Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val 195 200 205Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His 210 215 220Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val225 230 235113702DNAHomo sapiens 113atgagggtcc ctgctcagct cctggggctc ctgctgctct ggttcccagg tgccaggtgt 60gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 120atcacttgcc ggacaagtca gggcattaga aatgatttag gctggtatca gcagaaacca 180gggaaagccc ctaagcgcct gatctatgct gcatccagtt tgcaaagtgg ggtcccatca 240aggttcagcg gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct 300gaagattttg caacttatta ctgtctacag cataatagct accctcccac tttcggcgga 360gggaccaagg tggagatcaa acgaactgtg gctgcaccat ctgtcttcat cttcccgcca 420tctgatgagc agttgaaatc tggaactgct agcgttgtgt gcctgctgaa taacttctat 480cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 540gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 600ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 660ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 702114702DNAHomo sapiens 114acactctccc ctgttgaagc tctttgtgac gggcgagctc aggccctgat gggtgacttc 60gcaggcgtag actttgtgtt tctcgtagtc tgctttgctc agcgtcaggg tgctgctgag 120gctgtaggtg ctgtccttgc tgtcctgctc tgtgacactc tcctgggagt tacccgattg 180gagggcgtta tccaccttcc actgtacttt ggcctctctg ggatagaagt tattcagcag 240gcacacaacg ctagcagttc cagatttcaa ctgctcatca gatggcggga agatgaagac 300agatggtgca gccacagttc gtttgatctc caccttggtc cctccgccga aagtgggagg 360gtagctatta tgctgtagac agtaataagt tgcaaaatct tcaggctgca ggctgctgat 420tgtgagagtg aattctgtcc cagatccact gccgctgaac cttgatggga ccccactttg 480caaactggat gcagcataga tcaggcgctt aggggctttc cctggtttct gctgatacca 540gcctaaatca tttctaatgc cctgacttgt ccggcaagtg atggtgactc tgtctcctac 600agatgcagac agggaggatg gagactgggt catctggatg tcacacctgg cacctgggaa 660ccagagcagc aggagcccca ggagctgagc agggaccctc at 702115234PRTHomo sapiens 115Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp Phe Pro1 5 10 15Gly Ala Arg Cys Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser 20 25 30Ala Ser Val Gly Asp Arg Val Thr Ile Thr Cys Arg Thr Ser Gln Gly 35 40 45Ile Arg Asn Asp Leu Gly Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro 50 55 60Lys Arg Leu Ile Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser65 70 75 80Arg Phe Ser Gly Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser 85 90 95Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Leu Gln His Asn 100 105 110Ser Tyr Pro Pro Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg 115 120 125Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln 130 135 140Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr145 150 155 160Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser 165 170 175Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr 180 185 190Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys 195 200 205His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro 210 215 220Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225 230

Claims

1. (canceled)

2. (canceled)

3. (canceled)

4. (canceled)

5. (canceled)

6. (canceled)

7. (canceled)

8. (canceled)

9. (canceled)

10. (canceled)

11. (canceled)

12. (canceled)

13. (canceled)

14. (canceled)

15. (canceled)

16. (canceled)

17. (canceled)

18. (canceled)

19. (canceled)

20. (canceled)

21. (canceled)

22. (canceled)

23. (canceled)

24. (canceled)

25. (canceled)

26. (canceled)

27. (canceled)

28. (canceled)

29. (canceled)

30. (canceled)

31. (canceled)

32. (canceled)

33. (canceled)

34. (canceled)

35. (canceled)

36. (canceled)

37. (canceled)

38. (canceled)

39. (canceled)

40. (canceled)

41. (canceled)

42. A method of activating an endogenous activity of a human erythropoietin receptor in a mammal, the method comprising the step of administering to said mammal a therapeutically effective amount of an antibody or antibody fragment thereof to activate said receptor, wherein said antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of SEQ ID NO:1.

43. A method of modulating an endogenous activity of a human erythropoietin receptor in a mammal, the method comprising the step of administering to a mammal a therapeutically effective amount of the antibody or antibody fragment of claim 1 to modulate the activity of the receptor.

44. A method of treating a mammal suffering aplasia, the method comprising the step of administering to a mammal in need of treatment a therapeutically effective amount of an antibody or antibody fragment thereof to activate said receptor, wherein said antibody or antibody fragment thereof does not interact with a peptide having an amino acid sequence of SEQ ID NO:1.

45. A method of treating a mammal suffering aplasia, the method comprising the step of administering to a mammal in need of treatment a therapeutically effective amount of the antibody or antibody fragment of claim 1 to modulate the activity of the receptor.

46. (canceled)

47. (canceled)

48. (canceled)

49. (canceled)

50. (canceled)

51. (canceled)

52. (canceled)

53. (canceled)

54. (canceled)

55. (canceled)

56. (canceled)

57. (canceled)

58. (canceled)

59. (canceled)

60. (canceled)