Establishment of A Cell Strain for High Expression of Human-Mouse Chimeric Antibody against Human TNF-α and Analysis of Expressed Antibody

Objective To establish a cell strain for high expression of human-mouse chimeric antibody against human TNF-α.Methods Transfect CHO cells with the constructed eukaryotic expression vector of gene encoding human-mouse chimeric antibody against human TNF-α and screen positive clones by DHFR/MTX gene amplification strategy.Analyze the characteristic of expressed antibody by ELISA,Western blot and neutralization test in vitro and compare with that of parental murine McAb.Results A transfected cell line 2F5 for high expression of chimeric antibody against human TNF-α was established.After static culture of the transfected cells for 4 d,the expression level of gene engineering antibody was about 80 mg/L.Compared with its parental McAb F7/2,the expressed antibody showed good specificity and neutralizing activity,and its relative affinity was 2.1×10 -10 mol/L.Conclusion A gene engineering cell strain for high expression of chimeric antibody against human TNF-α was successfully established.The expressed antibody showed potential prospect in clinical application.