# US 2013/0157262 A1 · Method for Selecting Antibody-Producing Cell Line, and Kit Thereof

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- Number: US 2013/0157262 A1
- Document ID: US20130157262A1
- Status: Published application
- Kind: A1
- Application number: 13/643,952
- Filed: 2011-04-29
- Published: 2013-06-20
- Earliest claimed: 2010-04-29
- Assignees: Korea Research Institute of Bioscience and Biotechnology
- Inventors: Eun Gyo Lee; Hong Weon Lee; Yeon Gu Kim; Joon Ki Jung; Jung Oh Ahn; Chun Sug Kim; Hyeok Won Lee; Joo Hwan Lee; Hye Lym Lee
- Web page: https://patentgrep.com/patent/US20130157262A1

## CPC

- C12Q 1/025: {for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics (antimicrobial activity C12Q1/18)}
- C12N 15/1055: {Protein x Protein interaction, e.g. two hybrid selection}
- G01N 33/6854: {Immunoglobulins}
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)

## Abstract

Provided is a method for selecting antibody-producing cell lines using a split fluorescent protein, and a kit for selecting antibody-producing cell lines. By using the split fluorescent protein to select the antibody-producing cell lines, the antibody-producing cell lines can be easily detected by observing whether a single fluorescent color derived from reassembly of the split fluorescent proteins is expressed, which leads in a drastic reduction in selection time and cost required to select highly productive antibody-producing cell line.

## Claims

### Claim 1

A method for selecting antibody-producing cell lines, comprising:
transfecting a first expression vector and a second expression vector into cells, wherein the first expression vector comprises a sequence which encodes a first fragment of a fluorescent protein and a sequence which encodes a heavy chain of an antibody, and the second expression vector comprises a sequence which encodes a second fragment of the fluorescent protein and a sequence which encodes a light chain of the antibody; and
selecting the antibody-producing cell lines by confirming fluorescence derived from reassembly between the first fragment and the second fragment of the fluorescent protein.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the fluorescent protein is a green fluorescent protein (GFP), a red fluorescent protein (RFP), a blue fluorescent protein (BFP), a yellow fluorescent protein (YFP), a cyan fluorescent protein (CFP), or an enhanced fluorescent protein (EFP).

### Claim 3 (depends on 1)

The method according to claim 1, wherein one of the first fragment and the second fragment of the fluorescent protein is a C-terminal fragment of the fluorescent protein, and the other is an N-terminal fragment of the fluorescent protein.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the first expression vector further comprises a sequence which encodes a first linker peptide conjugated to the sequence which encodes the first fragment of the fluorescent protein,
the second expression vector further comprises a sequence which encodes a second linker peptide conjugated to the sequence which encodes the second fragment of the fluorescent protein, and
the first linker peptide and the second linker peptide are constructed so as to be conjugated to each other.

### Claim 5 (depends on 1)

The method according to claim 1, further comprising internal ribosome entry site (IRES) sequences between entry regions between the sequence which encodes the first fragment of the fluorescent protein of the first expression vector and an insertion site into which the sequence which encodes the heavy chain of the antibody are inserted; and between the sequence which encodes the second fragment of the fluorescent protein of the second expression vector and an insertion site into which the sequence which encodes the light chain of the antibody are inserted.

### Claim 6 (depends on 1)

The method according to claim 1, further comprising:
confirming production of the antibody by the selected antibody-producing cell lines.

### Claim 7

A kit for selecting antibody-producing cell lines, comprising:
a first expression vector comprising a sequence which encodes a first fragment of a fluorescent protein and an insertion site into which a sequence which encodes a heavy chain of an antibody may be introduced; and
a second expression vector comprising a sequence which encodes a second fragment of the fluorescent protein and an insertion site into which a sequence which encodes a light chain of the antibody may be introduced.