Volume 1 Issue 4
Dec.  2017
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Huan Wang, Xiaojun Tang, Xin Xu, Chu Chu, Siping Xiong, Feng Zheng, Qisong Peng. The establishment of human anti-TROP2 antibody IgG and its inhibition function on pancreatic cancer cell proliferation[J]. Blood&Genomics, 2017, 1(4): 65-71. doi: 10.46701/APJBG.2017042017045
Citation: Huan Wang, Xiaojun Tang, Xin Xu, Chu Chu, Siping Xiong, Feng Zheng, Qisong Peng. The establishment of human anti-TROP2 antibody IgG and its inhibition function on pancreatic cancer cell proliferation[J]. Blood&Genomics, 2017, 1(4): 65-71. doi: 10.46701/APJBG.2017042017045

The establishment of human anti-TROP2 antibody IgG and its inhibition function on pancreatic cancer cell proliferation

doi: 10.46701/APJBG.2017042017045
  • Publish Date: 2017-12-30
  • On the basis of anti-TROP2 Fab antibody, this study seek to construct a eukaryotic expression system of human anti-TROP2 antibody IgG, and to analyse the inhibition function of human anti-TROP2 antibody IgG in the cell proliferation of pancreatic cancer. The heavy and light chain genes of anti-TROP2 antibody were amplified respectively to establish the recombinant expression vector of human anti-TROP2 antibody IgG, named pWS-anti-TROP2. The human anti-TROP2 antibody IgG was obtained through transfecting the plasmids into the CHO dhfr- cell line, selecting the monoclonal cell strains with high amounts of antibody expression by MTX screening and applying Protein G affinity in purification. The identification and immunologic activity of human anti-TROP2 antibody IgG were researched by Western Blot,SDS-PAGE, ELISA, immunofluorescence assay and flow cytometry method (FCM). MTT assay was conducted to analyse the inhibition effect of human anti-TROP2 antibody IgG on BxPC3 cell proliferation. The human anti-TROP2 antibody IgG eukaryotic expression system was established successfully to express human anti-TROP2 antibody IgG, in which the molecular weight of heavy chain and light chain were consistent with expectation, and it could specifically combine with TROP2 protein, the antibody titer reached 1:6,400. The MTT assay results indicated that human anti-TROP2 antibody IgG had a significant effect on inhibiting the proliferation of BxPC3 cell, and the inhibition function can be gradually increased with improved antibody dose and prolonged time. In the study, the human anti-TROP2 antibody IgG eukaryotic expression system was constructed successfully, the antibody could specifically bind to TROP2 protein on the surface of pancreatic cancer cells, and it is shown to have a significant inhibitory action in pancreatic cancer cell proliferation.

     

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