中国肿瘤生物治疗杂志2011，Vol.18Issue(1)：11-16,6.DOI:10.3872/j.issn.1007-385X.2011.01.003
EGFRvⅢ特异性单链抗体的制备及其靶向性能
Preparation of EGFRVⅢ specific-chain Fv and its targeting activity
摘要
Abstract
Objective: To screen for EGFRvⅢ specific single-chain Fv (scFv) by phage display library and to examine its targeting activity. Methods: EGFRv Ⅲ specific scFv phage library was constructed, and the positive EGFRv Ⅲ-scFv clone was screened by ELISA. After cloned into pCANTAB-Thrombin-His vector, EGFRv Ⅲ-scFv plasmid was transformed into E. coli HB2151, and soluble EGFRv Ⅲ-scFv was induced by IPTG. The specific binding activity of EGFRv Ⅲ-scFv with EGFRv Ⅲ was studied by indirect immunofluorescence and in vivo imaging. Results: An EGFRv Ⅲ-scFv phage library was successfully constructed and 16 EGFRv Ⅲ-scFv positive clones were identified by ELISA. One clone named EGFRv Ⅲ-scFv-2A1 was re-cloned into pCANTAB-Thrombin-His vector and soluble EGFRv Ⅲ-scFv-2Al was successfully obtained. EGFRv Ⅲ-scFv-2A1 could specifically bind with HuH7-EGFRv Ⅲ and HuH7 hepatoma cells, but not with HuH7-EGFR and HuH7 cells in vitro. In vivo, fluorescence-labeled EGFRv Ⅲ-scFv-2A1 could only bind with U87MG-EGFRv Ⅲ glioma cells implanted tumor tissues, but not with that of U87MG cells implanted ones. Conclusion: The prepared EGFRv Ⅲ-scFv-2A1 can specifically bind with EGFRv Ⅲ, and it might be used for diagnosis and targeted therapy of tumors.关键词
EGFRvⅢ/scFv/脑胶质瘤/肝癌/移植瘤/靶向治疗/活体成像分类
医药卫生引用本文复制引用
张庆丽,石必枝,蒋华,周敏,王海,孔娟,谢海龙..EGFRvⅢ特异性单链抗体的制备及其靶向性能[J].中国肿瘤生物治疗杂志,2011,18(1):11-16,6.基金项目
卫生部"重大新药创制"科技重大专项资助项目(No.2009ZX09103-701). （No.2009ZX09103-701）
