Preparation and in vitro Bioactivity Characterization of Recombinant Humanized Anti-MSLN Monoclonal Antibody
Mesothelin(MSLN)could be served as an important target for immunotherapy for its wide expression in a variety of solid tumors and less expression in normal mesothelial cells.In this study,a recombinant humanized IgGl monoclonal antibody(rhMSLN mAb)sequence against MSLN with high affinity was constructed into the mammalian cell expression vector pcDNA3.4,and efficiently expressed by the Expi293F cell line.Then,the affinity of the antibody was assessed by surface plasmon resonance and flow cytometry at the molecular and cellular levels respectively.Lactate dehydrogenase(LDH)release assay was used to identify the antibody-dependent cellular cytotoxicity(ADCC)mediated by the antibody.The results showed that rhMSLN mAb could yield up to 70.2 mg/L under specific conditions by the Expi293F cell line.The affinity to recombinant MSLN was 7.02 nmol/L.The EC50 of ADCC against human gastric carcinoma cell line(NCI-N87 cell)was 2.48×10-3 μg/mL.This study provided explorative support for the development and production of biopharmaceuticals targeting MSLN.
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