安徽医科大学学报2018，Vol.53Issue(2)：176-181,6.DOI:10.19405/j.cnki.issn1000-1492.2018.02.003
全人源化PD-L1单抗功能活性及其对人肺腺癌HCC827细胞毒性作用的研究
Functional activity of humanized PD-L1 monoclonal antibody and its toxic effects on human lung adenocarcinoma cell line HCC827
摘要
Abstract
Objective To explore the function and activity of human humanized PD-L1 monoclonal antibody and its toxicity to human lung adenocarcinoma cell line HCC827. Methods Sodium dodecyl sulfate polypropylene gel electrophoresis (SDS-PAGE) was used to determine the purity of Atezolizumab and PD-L1 monoclonal antibodies. The binding and competitive blocking activity of the self-developed PD-L1 monoclonal antibody were determined by enzyme linked immunosorbent assay (ELISA) with Atezolizumab as a positive control. Cell blocking activity of two antibody was measured by cell interruption test. Flow cytometry (FACS) was used to detect the expression of PD-L1 on the surface of human lung adenocarcinoma cell line HCC827, antibody dependent cell-mediated cytotoxicity(ADCC) effects were detected in two of antibodies. Results High purity Atezolizumab and PD-L1 monoclonal antibodies were identified by SDS-PAGE. Cell blocking activity of two antibody was measured by cell interruption test. ELISA showed that both PD-L1 monoclonal antibodies and Atezolizumab could bind to human PD-L1 and block the binding of PD-L1 to PD-1. Both PD-L1 monoclonal antibodies and Atezolizumab could block the binding of two cell surface molecules PD-L1 with PD-1, EC50 were 0. 208 5 μg/ml and 0. 154 8 μg/ml, respectively, there was no significant differences in certain concentration. FACS results showed that high expression of PD-L1 in human lung adenocarcinoma HCC827 cells. Cytotoxicity experiments showed that when the ratio between effector cells and target cells was constant, monoclonal antibodies against PD-L1 and Atezolizumab have strong toxic effects on human lung adenocarcinoma cell line HCC827. Conclusion The research of humanized monoclonal antibody PD-L1 has functional activity, T cell receptor signaling can be attenuated by blocking the combination of PD-L1 and PD-1 on the cell surface, and may exert a certain antitumor effect through the ADCC effect.关键词
PD-L1单抗/阿特朱单抗/ADCC/肺癌Key words
PD-L1 antibody/Atezolizumab/antibody dependent cell-mediated cytotoxicity/lung cancer分类
医药卫生引用本文复制引用
谢庆书,束军,陈晴晴,胡思怡..全人源化PD-L1单抗功能活性及其对人肺腺癌HCC827细胞毒性作用的研究[J].安徽医科大学学报,2018,53(2):176-181,6.基金项目
安徽省自然科学基金面上项目(编号:1308085MH141) （编号:1308085MH141）
安徽高校省级自然科学研究项目(编号:KJ2011A173) （编号:KJ2011A173）
