Preliminary Study On Anti-NRP-1b1b2Monoclonal Antibody Purification, Identification And Activity Of Anti-tumor Activity In Gastric Cancer Cells

ObjectivesTo study the expression and location of neuropilin-1(NRP-1) in human gastric cancercells; and to investigate the effect of the anti NRP-1monoclonal antibody (mAb) oncell growth, proliferation and migration of gastric cancer cell lines..MethodsFirstly, the anti-NRP-1mAb were obtained by rProtein A column purification fromthe ascites. the affinity of antibody was identified by ELISA method and identified bySDS-PAGE purity antibodies, flow cytometry was used to observe combinationability of anti-NRP-1mAb to BGC823and MKN28cells, immunofluorescence wasused to observe the localization of anti-NRP-1mAb in the cells, respectively. Finally,Then we used cell proliferation experiment and wound healing experiment to researcheffects of anti-NRP-1mAb on cell proliferation, migration of BGC823andMKN28cells.ResultsOur experiment results showed that the purity of purified anti-NRP-1mAb was morethan95%, concentration was4mg/mL. Results of ELISA and Flow cytometryindicated that the anti-NRP-1mAb could bind to recombinant NRP-1b1b2proteinand gastric cancer cell line BGC823and MKN28with higher affinity.Immunofluorescence experiments results showed that the anti-NRP-1mAblocalizated on the cell surface. Cell proliferation experiment and wound healingexperiment respectively showed that anti-NRP-1mAb inhibited the growth, migrationof BGC823and MKN28cells.Conclusions(1) We built an affinity chromatography purification method to purify theanti-NRP-1mAb, anti-NRP-1mAb with higher affinity and purity was obtained. (2) Anti-NRP-1mAb inhibited the growth, migration of BGC823and MKN28cells.The Anti-NRP-1mAb is likely to be a prospective antibody drug candidate for tumortreatment.