Construction,expression,and the analysis of antitumor activity of all anthropogenic medicinal anti-EGFR ScFv (HL-L-L) / 中国生化药物杂志

ABSTRACT

Objective To construct,express and analysis the antitumor activity of anti-EGFR ScFv(HL-L-L),and analyze bioinformatics and characterize correlation spectroscopy.Methods EGFR ScFv(HL-L-L)gene was obtained by OVER LAPING technology and construction of prokaryotic expression vector pET-28a-c(+)-VH-L-L.After induced expression,target protein HL-L-L was purified by Ni column,and its tertiary structures was predicted with SWISS - MODEL;size distribution and zeta potential distribution of the target protein HL-L-L were characterized by NanoZSP;the inhibition effects of HL-L-L on breast cancer cell proliferation were studied with the method of pharmacology.Results Prokaryotic expression vector pET-28a-c(+)-VH-L-L was constructed successfully,high purity of protein HL-L-L was obtained,and successfully predicted its three-dimensional structure;the size was about 489.7 nm,and the Zeta potential was 38.1 ,negatively charged.It had a certain inhibition effect on MCF # 7 proliferation,compared with blank group,and the difference was significant(P<0.01 ).Conclusion This study successfully express all anthropogenic medicinal anti-EGFR ScFv(HL-L-L),which has significant inhibiting effect on breast cancer cell proliferation.It provides the foun dation for further development of small molecules of anti-EGFR antibody drug.