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抗癌胚抗原重组单链Fv抗体片段的制备、纯化及特性分析

Production, purification and characterization of an anti-(carcinoembryonic antigen) recombinant single-chain Fv antibody fragment.

作者信息

Pérez L, Vázquez J E, Ayala M, Siccardi A G, Tresoldi E, de Lalla C, Gavilondo J V, Sidoli A

机构信息

Department of Biological and Technological Research, San Raffaele Scientific Institute, Milano, Italy.

出版信息

Biotechnol Appl Biochem. 1996 Aug;24(1):79-82.

PMID:8756397
Abstract

Specific targeting of radioactive agents to tumour cells has been the main objective of the in vivo use of monoclonal antibodies and their fragments. In particular, specific antibodies to carcinoembryonic antigen (CEA)-expressing tumours can be used either for diagnosis or therapy, if targeting could be improved. The expression of antibody fragments in bacteria allows the preparation of engineered molecules with antigen-binding properties and a better penetration into the tumour. A specific anti-CEA single-chain Fv fragment was produced in bacteria and purified. Its binding activity has been demonstrated in ELISA, immunocytochemistry, immunohistochemistry, fluorescence-activated cell sorting and the kinetic parameters determined by the plasmon surface resonance.

摘要

将放射性制剂特异性靶向肿瘤细胞一直是单克隆抗体及其片段体内应用的主要目标。特别是,如果能改进靶向性,针对表达癌胚抗原(CEA)肿瘤的特异性抗体可用于诊断或治疗。在细菌中表达抗体片段能够制备出具有抗原结合特性且能更好地渗透到肿瘤中的工程分子。一种特异性抗CEA单链Fv片段在细菌中产生并纯化。其结合活性已在酶联免疫吸附测定、免疫细胞化学、免疫组织化学、荧光激活细胞分选以及通过表面等离子体共振测定的动力学参数中得到证实。

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