US 12,286,620 B2
Antibody library construction method and application thereof
Yongchao Yao, Guangdong (CN); Youjia Li, Guangdong (CN); Sha Yin, Guangdong (CN); and Guangjie Liu, Guangdong (CN)
Assigned to SHENZHEN ISTIRBIO CO., LTD., Guangdong (CN)
Appl. No. 16/973,722
Filed by SHENZHEN ISTIRBIO CO., LTD., Guangdong (CN)
PCT Filed Sep. 26, 2019, PCT No. PCT/CN2019/108128
§ 371(c)(1), (2) Date Dec. 9, 2020,
PCT Pub. No. WO2020/125120, PCT Pub. Date Jun. 25, 2020.
Claims priority of application No. 201811571758.5 (CN), filed on Dec. 21, 2018.
Prior Publication US 2021/0340523 A1, Nov. 4, 2021
Int. Cl. C12N 15/10 (2006.01); C07K 16/28 (2006.01); C07K 16/30 (2006.01); C40B 30/04 (2006.01)
CPC C12N 15/1037 (2013.01) [C07K 16/2803 (2013.01); C07K 16/303 (2013.01); C40B 30/04 (2013.01)] 10 Claims
 
1. A method for screening antibody expression cells using drugs, comprising:
(1) transfecting a vector containing a CIS activator and a screening marker gene combination into a cell and constructing a monoclonal cell line stably transfected with the CIS activator and the screening marker gene combination, wherein the screening marker gene combination comprises a combination of a drug resistance gene, a suicide gene, and a fluorescent protein gene or a molecular tag;
(2) transfecting antibody gene library expression vectors carrying an extracellular antibody library coding domain, a Notch core domain and an intracellular transcription domain into cells of the monoclonal cell line of the step (1), to obtain a library of antibody expression cells;
(3) expressing the antibody expression cells of step (2), contacting them with an antigen, and adding a screening drug for the drug resistance gene for positive screening of the antibody library expression cells; and
(4) adding a screening drug for the suicide gene for negative screening of the screened cells in step (3), to obtain a screened library of antibody expression cells.