U.S. patent application number 09/728317 was filed with the patent office on 2001-11-15 for method for producing micro-carrier and test method by using said micro-carrier.
This patent application is currently assigned to GeneMaster Biotechnology Corp.. Invention is credited to Chang, Rong-Seng, Chao, Yu-Chan.
Application Number | 20010041369 09/728317 |
Document ID | / |
Family ID | 21659696 |
Filed Date | 2001-11-15 |
United States Patent
Application |
20010041369 |
Kind Code |
A1 |
Chang, Rong-Seng ; et
al. |
November 15, 2001 |
Method for producing micro-carrier and test method by using said
micro-carrier
Abstract
The invention provides a method for producing a micro-carrier,
which includes patterning pluralities of bar code on a mask;
exposing the bar code to a substrate coated with photoresist;
etching and removing residual photoresist and electroforming to a
nickel plate; placing a bead coated with biotin or poly-L-lysine
between two-nickel plates, and compressing the bar code on the
surface of the bead to form a microcake-like particle with bar
code; and combining the particle with the corresponding
bio-molecule thereof to produce a micro-carrier with a label. The
invention also provides a test method for identifying a
bio-molecule, which includes mixing several micro-carriers with the
labeled unknown bio-molecules; and identifying the bar code on the
micro-carrier via image recognition system, wherein the numbers and
types of the known micro-carrier can be flexibly adjusted.
Inventors: |
Chang, Rong-Seng; (Taipei,
TW) ; Chao, Yu-Chan; (Taipei, TW) |
Correspondence
Address: |
DARBY & DARBY P.C.
805 Third Avenue
New York
NY
10022
US
|
Assignee: |
GeneMaster Biotechnology
Corp.
|
Family ID: |
21659696 |
Appl. No.: |
09/728317 |
Filed: |
December 1, 2000 |
Current U.S.
Class: |
436/518 |
Current CPC
Class: |
B01J 19/0046 20130101;
B01J 2219/00502 20130101; B01J 2219/00702 20130101; Y10S 435/97
20130101; Y10S 436/823 20130101; B01J 2219/00554 20130101; B01J
2219/00725 20130101; B01J 2219/0074 20130101; Y10S 436/805
20130101; Y10T 436/25375 20150115; B01J 2219/00545 20130101; G03F
7/0017 20130101; Y10S 435/961 20130101; G01N 33/54313 20130101;
Y10S 435/969 20130101; C40B 70/00 20130101; B01J 2219/00547
20130101; G01N 33/552 20130101; C40B 40/10 20130101; B01J
2219/00722 20130101; B01J 2219/005 20130101; B01J 2219/00596
20130101; C40B 40/06 20130101; Y10T 436/11 20150115 |
Class at
Publication: |
436/518 |
International
Class: |
G01N 033/543 |
Foreign Application Data
Date |
Code |
Application Number |
May 12, 2000 |
TW |
89109106 |
Claims
What is claimed is:
1. A method for producing a micro-carrier of bio-molecule,
comprising: preparing a bead and coating said bead with a layer of
bio-molecule binding material; patterning pluralities of bar code
on a mask using an integrated circuit process, wherein the bar code
represents the desired bio-molecule; exposing the bar code to a
substrate coated with photoresist using photolithography; etching
and removing residual photoresist, wherein the bar code is formed
on the substrate, and electroforming to a nickel plate; placing
said bead between two-nickel plates, and compressing the bar code
on the surface of said bead to form a microcake-like particle with
a bar code; coating a layer of bio-molecule binding material onto
said particle; and combining said particle with the corresponding
bio-molecule thereof to produce a micro-carrier with a label.
2. The method as claimed in claim 1, wherein said particle is
formed from the beads placed between two-nickel plates by dropping
a UV photosenstizer micelle onto the nickel plates, followed by UV
irradiation for curing.
3. The method as claimed in claim 1, wherein said microcake-like
particle is produced by simultaneously patterning a cake-like
pattern and bar code on a mask, etching to form a mold, and molding
by injection or hot compression.
4. The method as claimed in claim 1, wherein said micro-carrier is
further characterized by the shape, size, or color of said
micro-carrier as a code.
5. The method as claimed in claim 1, wherein the bio-molecule
comprises nucleic acid, oligonucleotide, peptide nucleic acid,
antigen, antibody, enzyme or protein.
6. The method as claimed in claim 1, wherein the bio-molecule
binding material comprises biotin or poly-L-lysine.
7. A test method for identifying a bio-molecule, comprising:
providing a vial containing at least one micro-carrier with a bar
code; adding a labeled unknown bio-molecule to said vial and
mixing, wherein a signal is obtained when said micro-carrier is
complementary in combination with the unknown bio-molecule;
transferring said micro-carrier in the vial onto a transporter,
wherein a microscope connected with a computer is set above the
transporter; and identifying said bar code of the signaling
micro-carrier by an image recognition system, thereby identifying
the unknown bio-molecule.
8. The test method as claimed in claim 5, wherein the bio-molecule
comprises nucleic acid, oligonucleotide, peptide nucleic acid,
antigen, antibody, enzyme or protein.
9. The test method as claimed in claim 5, wherein the bio-molecule
binding material comprises biotin or poly-L-lysine.
Description
BACKGROUND OF THE INVENTION
[0001] 1. Field of the Invention
[0002] The present invention relates to a method for preserving and
testing biologically genetic information, and particularly to a
micro-carrier and a test method for identifying DNA, proteins and
other complementary substances by using a bar code labeled
micro-carrier.
[0003] 2. Description of the Related Arts
[0004] Biotechnology has been developing quickly in recent years.
Various products can be produced using molecular biology,
biological cells, or other metabolites thereof by this technique,
which can be extensively applied in the fields of pharmaceutical,
pesticide, environmental protection, process development, and
aquaculture.
[0005] The combination of biotechnology with electric technology is
a trend; wherein the most attractive is the biochip and DNA chip
(i.e. gene chip). In addition to silicon, the material of those
chips can include absorbent materials such as glass, plant
cellulose, gel, and organic polymers. The gene chip has various
gene fragments neatly aligned and adhered onto a nail-sized chip,
in which thousands upon thousands of gene fragments are
accommodated. Users can select different kinds of gene chips based
on their purposes.
[0006] The principle of the aforementioned gene chip is that
different groups of gene fragments are adhered onto a chip,
followed by immersion into a solution containing unknown genes
labeled with fluorescence. If the fluorescence-labeled gene matches
the specific gene fragment on the chip, a fluorescent signal
retained thereon due to complementary combination will be observed
by microscopy. Therefore, the unknown gene can be identified by the
complementary sequence adhered on the chip.
[0007] Under the design of large production, thousands upon
thousands of gene fragments or proteins are adhered onto the chip;
however, it has to avoid inaccuracy resulting from different gene
fragments or proteins whose locations on the chip are too close.
Thus, the precise control of the spots on the chip is very
important. Moreover, the precise control requires expensive
equipment, which restricts the application of the chip. Therefore,
there is still a need for developing a bio-molecule database and
test technique thereof, which possess advantages of more
efficiency, low cost, and low limitation.
SUMMARY OF THE INVENTION
[0008] It is therefore the main purpose of the present invention to
provide a convenient, inexpensive, and rapid method for producing a
micro-carrier of bio-molecule (e.g. gene or protein), and a method
for testing bio-molecules by using the micro-carrier.
[0009] Another purpose of the present invention is to provide a
test method for identifying a bio-molecule, wherein the numbers and
types of the known micro-carrier can be flexibly adjusted.
[0010] According to the method of the present invention, bar codes
are patterned on a mask using an integrated circuit process,
followed by exposure to a substrate coated with photoresist using
photolithography. After etching and removing residual photoresist,
the desired bar code can be formed on the substrate, and
subsequently a nickel plate is thus electroformed. Before or after
coating with bio-molecule binding material, a bead (Q-bot) is
placed between two-nickel plates, and the bar code is then hot
compressed onto the surface of the bead to form a microcake-like
particle with bar code. Afterwards, each of the particles mentioned
above are combined with the corresponding genes or proteins thereof
to produce large amount of micro-carriers with labels. On the other
hand, according to the method for testing bio-molecules described
herein, large amounts of micro-carriers mentioned above are
employed and the labeled (for example, fluorescence-labeled)
unknown bio-molecules are mixed with the micro-carriers. The
hybridization intensities of the fluorescence or different markers
of the unknown bio-molecules thus are identified by the bar code on
the micro-carrier via an image recognition system.
BRIEF DESCRIPTION OP THE DRAWINGS
[0011] The present invention will be more fully understood and
further advantages will become apparent when reference is made to
the following description of the invention and the accompanying
drawings in which:
[0012] FIG. 1 is a diagram showing an insert for producing the
micro-carrier of the present invention; and
[0013] FIG. 2 is a schematic diagram showing the process for
producing the micro-carrier of the present invention.
DETAILED DESCRIPTION OF THE INVENTION
[0014] The present invention is characterized by the combination of
biotechnology with integrated circuit process to produce a
bio-molecule micro-carrier. Another feature of the present
invention is a method for testing unknown bio-molecules by using
the micro-carrier.
[0015] The method for producing a micro-carrier of the present
invention is performed as follows. A layer of bio-molecule binding
material, such as biotin, poly-L-lysine, etc., was coated onto the
surface of a bead. The desired individual bio-molecules (e.g. gene
or protein) were represented by a corresponding bar code, wherein
pluralities of the bar code were patterned on a mask using an
integrated circuit process, followed by exposing to a substrate
coated with photoresist using photolithography. After etching and
removing residual photoresist, the bar code was formed on the
substrate, and subsequently a nickel plate was thus electroformed.
The aforementioned bead was placed between two-nickel plates, and
the bar code facing inwards was then hot compressed onto the
surface of the bead to form a microcake-like particle with the bar
code. A layer of bio-molecule binding material was coated onto the
particle before or after bar code patterning. Finally, the
particles mentioned above were combined with the corresponding
bio-molecules thereof to produce various micro-carriers of
bio-molecules with labels. Therefore, users can produce a vial
containing various micro-carriers with bar codes in accordance with
the present invention.
[0016] The term "micro-carrier" used herein refers to a bead marked
with a specific bar code, then coated with a layer of bio-molecule
binding material, and then carries a corresponding bio-molecule.
The material of the bead is not limited, including silicon, glass,
plant cellulose, gel, and organic polymers. The size of the bead
ranges from 20 .mu.m to 200 .mu.m in diameter, preferably less than
100 .mu.m.
[0017] The bio-molecules used herein can include, but are not
limited to, nucleic acid, oligonucleotide, peptide nucleic acid
(PNA), antigen, antibody, enzyme or protein.
[0018] In the process of producing the above particles, the
hemisphere particles can be alternatively formed from the beads
placed between two-nickel plates by dropping a UV photosensitizer
micelle, such as Arabic micelle, onto the nickel plates, followed
by UV irradiation for curing.
[0019] In addition, the cake-like pattern and bar code can be
simultaneously patterned on a masks, as shown in FIG. 1, followed
by etching to form a mold. The microcake-like particle can thus be
molded by injection or hot compression.
[0020] Another aspect of the present invention provides a method
for testing an unknown bio-molecule by using the micro-carrier
mentioned above. The method is comprised of the following steps:
providing a vial containing numerous micro-carriers with bar code;
adding a labeled (for example, fluorescence-labeled) unknown
bio-molecule to said vial and mixing (i.e. hybridizing), wherein a
signal, such as fluorescence, is obtained when the micro-carrier is
complementary with or recognized by the unknown bio-molecule;
transferring the micro-carrier in the vial onto a transporter,
wherein a microscope connected with a computer is set above the
transporter; and identifying said bar code of the signaling
micro-carrier by an image recognition system, thereby identifying
the unknown bio-molecule.
[0021] In addition to the bar code used to identify bio-molecules,
the present invention further employs the shape, size, color, etc.
of the carrier as codes, which can be classified into many
categories, such as: (1) Shape. The sphere bead described above can
be replaced by a rectangle or polygon. For example, a certain kind
of length and width can represent a specific bio-molecule, or
either a triangle or polygon with sides of different length can
represent different bio-molecules. (2) Size. For example, the large
bead represents one bio-molecule and the small one represents
another. The diameter of the bead can be used as a bio-molecule
marker. (3) Color Different colors can represent different
bio-molecules. For example, red, yellow, blue, and white can be
used to represent four different kinds of bio-molecules. Similarly,
each micro-carrier can be identified and counted via the microscope
connected with computer and the image recognition system.
[0022] Furthermore, the insert 10 of the aforementioned carrier
with different shape and/or size can also be produced by
photolithography, as shown in FIG. 1, followed by injection or hot
compression. The resulting particles 12 are wedged in insert 10 due
to their very small size. The insert 10 can be electrified with a
negative charge and the particles 12 can thus be attracted to a
collection plate 14
* * * * *