High-Level Expression of Anti-Vascular Endothelial Growth Factor Chimeric Antibody in Eukaryotic Cells
10.3321/j.issn:1000-467X.2001.03.013
- VernacularTitle:抗人血管内皮生长因子嵌合抗体在真核细胞中的高效表达
- Author:
Yu-Liang RAN
1
;
Zhi-Hua YANG
;
Li-Xin SUN
;
Long YU
;
Jun LIU
;
Zhi-Wei DONG
Author Information
1. Peiking Union Medical College
- From:Chinese Journal of Cancer
2001;20(3):282-286
- CountryChina
- Language:Chinese
-
Abstract:
Objective：This study was designed to express chimeric anti-VEGF (vascular endothelial growth factor) antibody in dihydrofolate reductase-deficient Chinese hamster ovary (CHO-dhfr-)cells at high-level, and explore an optimum method to obtain high-level expression cells clone. Methods：The light chain and heavy chain genes of chimeric anti-VEGF antibody were induced into CHO-dhfr-cells using a novel eukaryotic high-level expression vectors system for genetic engineering antibodies. High-level expression was achieved after subcloning and several rounds of co-amplification of methotrexate (MTX). Biological features and productive amount of chimeric antibody was charactered by ELISA. Result：The cells strain that secret anti-VEGF chimeric antibody at the highest level of 28 μ g/ml was established. The cells were subcloned following each round of co-amplification of MTX, while greatly different results were obtained using three methods. The chimeric antibody contained constant regions of human immunoglobin and had the specificity against VEGF by ELISA. Conclusion：The anti-VEGF mouse-human chimeric antibody was expressed at high-level successfully in CHO cells. This may be an optimum method to obtain high-level expression cells clone for the eukaryotic high-level expression vectors system.
