Disulfide-stabilized single-chain antibody-targeted superantigen: Construction of a prokaryotic expression system and its functional analysis  

作　　者：Jian-Li Wang Yu-Ling Zheng RU Ma Bao-Li Wang Ai-Guang Guo Yong-Qiang Jiang 

机构地区：[1]Department of Life Science, Northwest Sci-Tech University of Agriculture and Forestry,No. 22 Xi nong Road, Yangling 712100, Shaanxi Province, China [2]Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, No. 20 Dongda Street, Fengtai District, Beijing 100071,China

出　　处：《World Journal of Gastroenterology》2005年第31期4899-4903,共5页世界胃肠病学杂志（英文版）

基　　金：Supported by the National Natural Science Foundation of China,No. 30271478

摘　　要：AIM: To construct the expression vector of B3 (scdsFv)-SEA (D227A) and to identify its binding and cytotoxic ability to B3 antigen positive carcinoma cell lines.METHODS: This fusion protein was produced by a bacterial expression system in this study. It was expressed mainly in the inclusion body. The gene product was solubilized by guanidine hydrochloride, refolded by conventional dilution method, and purified using SP-sepharose cation chromatography.RESULTS: The expression vector B3 (scdsFv)-SEA-PETwas constructed, the expression product existed mainly in the inclusion body, the refolding product retained the binding ability of the single-chain antibody and had cytotoxic effect on HT-29 colon carcinoma cells. The stability assay showed that the resulting protein was stable at 37 ℃.CONCLUSION: This genetically engineered B3 (scdsFv)-SEA fusion protein has bifunction of tumor targeting and tumor cell killing and shows its promises as an effective reagent for tumor-targeted immunotherapy.AIM： To construct the expression vector of B3 （scdsFv）-SEA （D227A） and to identify its binding and cytotoxic ability to B3 antigen positive carcinoma cell lines. METHODS： This fusion protein was produced by a bacterial expression system in this study. It was expressed mainly in the inclusion body. The gene product was solubilized by guanidine hydrochloride, refolded by conventional dilution method, and purified using SP-sepharose cation chromatography. RESULTS： The expression vector B3 （scdsFv）-SEA-PET was constructed, the expression product existed mainly in the inclusion body, the refolding product retained the binding ability of the single-chain antibody and had cytotoxic effect on HT-29 colon carcinoma cells. The stability assay showed that the resulting protein was stable at 37 ℃. CONCLUSION： This genetically engineered B3 （scdsFv）-SEA fusion protein has bifunction of tumor targeting and tumor cell killing and shows its promises as an effective reagent for tumor-targeted immunotherapy.

关 键 词：B3 monoclonal antibody Single-chain disulfide-stabilized Fv SUPERANTIGEN 

分 类 号：R730[医药卫生—肿瘤]