Selection and Characterization of Human Anti-MAGE-A1 scFv and Immunotoxin
Selection and Characterization of Human Anti-MAGE-A1 scFv and Immunotoxin
复制标题
人抗 MAGE-A1 scFv 和免疫毒素的选择和表征。
DOI：
10.2174/18715206113139990134
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发表时间：
2013-10-01
影响因子：
2.8
通讯作者：
Chen, Ren-Jie
中科院分区：
文献类型：
--
作者：
Lin, Hong;Mao, Yuan;Chen, Ren-Jie
黑色素瘤相关抗原(MAGE)表达于多种肿瘤细胞表面，是通过抗MAGE-A1抗体进行生物治疗的靶点。本研究构建了人源单链可变区(ScFv)噬菌体抗体库，并将其应用于MAGE-A1包被的重组免疫管中。用固定化金属螯合亲和层析(IMAC)表达和纯化了可溶性抗MAGE-A1单链抗体。经酶联免疫吸附试验、斑点印迹和免疫沉淀分析证实，抗MAGE-A1单链抗体能与天然MAGE-A1结合。用iMac成功表达和纯化了该免疫毒素。结果表明，人源性抗MAGE-A1免疫毒素可为临床肿瘤治疗提供有价值的药物。
Melanoma-associated antigen (MAGE) is expressed on the surface of multiple tumor cell types and is a promising target of biotherapeutic drug delivery via the anti-MAGE-A1 antibody. In this study, a human single-chain variable fragment (scFv) antibody phage library was generated and applied to recombinant MAGE-A1-coated immunotubes by phage display technology. The soluble anti- MAGE-A1 scFv was expressed and purified by immobilized metal-chelated affinity chromatography (IMAC). The anti-MAGE-A1 scFv could bind native MAGE-A1 confirmed by enzyme-linked immunosorbent assay (ELISA), dot blot, and immunoprecipitation (IP) analysis. The immunotoxin was expressed and purified by IMAC successfully. The results indicated that the human anti-MAGE-A1 immunotoxin could provide a valuable drug for clinic cancer therapy.