Preparation of site-specific antibody-drug conjugate targeting glypican 3 and evaluation of its anti-tumor activity
10.13200/j.cnki.cjb.004000
- VernacularTitle:抗磷脂酰肌醇蛋白聚糖3抗体定点偶联药物的制备及其抗肿瘤活性评价
- Author:
SUN Zhaopeng
- Publication Type:Journal Article
- From:
Chinese Journal of Biologicals
2023;36(10):1192-1197
- CountryChina
- Language:Chinese
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Abstract:
Objective To obtain monoclonal antibody against glypican 3(GPC3) by hybridoma technique and prepare antibody-drug conjugate(ADC) to investigate the anti-tumor activity of ADC.Methods Monoclonal antibody against GPC3with high affinity was obtained by immunizing male BALB/c mice and detected for its affinity to antigen and endocytosis rate in tumor cells by ELISA and flow cytometry.ADC drugs with DAR(drug-to-antibody ratio) of 2 were obtained by sitespecific enzyme coupling technique,and the inhibitory effects on proliferation of tumor cells HepG2 and Hep3b were detected by cell killing experiments.Results Antibody 16C8-8E6 had a high affinity at protein level with the EC_(50) of 2.51 ng/mL,and the affinity at cell level(stable strains 4E1,Hep3b and HepG2 with high expression of GPC3) was significantly higher than that of the original antibody GC33(t=14.9,13.0 and 12.9,respectively,each P <0.05).The fluorescence intensity of stable strain 4E1 with high expression of GPC3 was 20 542±107;The endocytosis rate was better than that of the original antibody GC33,which reached 73.9% within 48 h.16C8-8E6-ADC showed certain inhibitory activity on the proliferation of HepG2 and Hep3b cells.Conclusion The monoclonal antibody targeting GPC3 was successfully obtained,which lays a foundation of the research of immunotherapy targeting GPC3.
