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Title 

New cell line development for antibody-producing Chinese hamster ovary cells using split green fluorescent protein

 

분열형광 단백질을 이용한 신규 치료용 항체 생산 세포주 선별법

Authors 

Yeon Gu KimByoung-Woo ParkJungoh AhnJoon Ki JungHong-Weon LeeEun Gyo Lee

Publisher 

BioMed Central

Issue Date 

2012

Citation 

BMC Biotechnology, vol. 12, no. 1, pp. 24-24

Keywords 

Antibody productionCell line developmentCHO cellsFACSSplit GFP

Abstract 

Background: The establishment of high producer is an important issue in Chinese hamster ovary (CHO) cell culture considering increased heterogeneity by the random integration of a transfected foreign gene and the altered position of the integrated gene. Fluorescence-activated cell sorting (FACS)-based cell line development is an efficient strategy for the selection of CHO cells in high therapeutic protein production. Results: An internal ribosome entry site (IRES) was introduced for using two green fluorescence protein (GFP) fragments as a reporter to both antibody chains, the heavy chain and the light chain. The cells co-transfected with two GFP fragments showed the emission of green fluorescence by the reconstitution of split GFP. The FACS-sorted pool with GFP expression had a higher specific antibody productivity (qAb) than that of the unsorted pool. The qAb was highly correlated with the fluorescence intensity with a high correlation coefficient, evidenced from the analysis of median GFP and qAb in individual selected clones. Conclusions: This study proved that the fragment complementation for split GFP could be an efficient indication for antibody production on the basis of high correlation of qAb with reconstitution of GFP. Taken together, we developed an efficient FACS-based screening method for high antibody-producing CHO cells with the benefits of the split GFP system.

ISSN 

1472-6750

Link 

http://dx.doi.org/10.1186/1472-6750-12-24

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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