文献类型：期刊文献

英文题名：Design and screening of a chimeric survivin-specific nanobody and its anticancer activities in vitro

作者：Zhang, Na[1,2];Guo, Hua[1,2];Zheng, Wenyun[3,4];Wang, Tianwen[1,2];Ma, Xingyuan[1,2]

机构：[1]East China Univ Sci & Technol, Sch Biotechnol, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[3]East China Univ Sci & Technol, Sch Pharm, Shanghai, Peoples R China;[4]East China Univ Sci & Technol, Shanghai Key Lab New Drug Design, Shanghai, Peoples R China

年份：2016

卷号：27

期号：9

起止页码：839

外文期刊名：ANTI-CANCER DRUGS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000384001000005)】；

基金：This work was supported by the Shanghai Pujiang Program (13PJD012), the National Natural Science Foundation (30873190, 31300660), the Science and technology innovation action plan of Shanghai (14431904300), and a foundation for young teacher from Education Ministry of China (20120074120027), and the National Science Research Project 'Significant New Drugs Created' of Eleventh Five-year Plan (2009ZX09103-693), partially supported by the Open Funding Project of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：antibody engineering; anticancers; phage display; survivin nanobody

摘要：Survivin is a strong inhibitor of apoptosis protein and a promising target for cancer prevention and treatment. Here, we report the design and preparation of novel chimeric nanobodies (Nbs) that could specifically bind to survivin. We screened the peptides from phage-displayed libraries (7-mer, 12-mer) for nonconserved sequences of complementarity-determining regions (CDRs) in the scaffold of the Nb. By a combination of the nonconserved sequences for CDRs, the corresponding chimeric Nbs (10 Nbs) were prepared with genetic operations. The antisurvivin Nb TAT-Nb4A (a fusion with cellular transduction peptide TAT) was found to be the most efficient antibody on the basis of the results from enzyme-linked immunosorbent assay, MTT, and flow cytometry when these nanobodies were tested with hepatoma carcinoma cell HepG2. TAT-Nb4A could inhibit the growth of HepG2 and promote cancer cell apoptosis significantly in a dose-dependent and time-dependent manner: the apoptosis rate reached 52.5% when the concentration of TAT-Nb4A was 120g/ml. Western blotting with cells expressing survivin showed that the prepared nanobody could efficiently bind to expressed survivin and blocked the signaling pathway in which survivin played a role. This study provided a convenient and feasible method of obtaining a novel specific Nb with the case of survivin as a good example.