抗A型肉毒毒素人源单链抗体融合蛋白的重组设计

以获得的抗A型肉毒毒素单链抗体为模板，进行融合改构，将人IgGl的Fc片段连接到ScFv的C端，在大肠杆菌中实现抗体融合蛋白ScFv—Fc的表达，表达量30％以上，蛋白以包含体形式存在，经过体外变复性的抗体融合蛋白ScFv．Fc，进行Protein G Sepharose柱亲和层析纯化，纯度达90％-95％。体外活性检测结果表明，重组抗体融合蛋白ScFvFc可以特异结合A型肉毒类毒素抗原，其相对亲和力近似于母本单链抗体，其稳定性高于母本单链抗体。

Recombinant Design of Fc Fusion Protein of Human ScFv Against BoNTa

Human ScFv against botulinum neurotoxin serotype A(BoNTa) was modified by fusing human IgG1 Fc to C terminal of ScFv. ScFv-Fc fusion protein was expressed at high level over 30% of total host cell proteins in E.coli. Recombinant protein existed in inclusion body form. Renatured ScFv-Fc was purified to 90%~95% by Protein G Sepharose column. In vitro ScFv-Fc could bind specific to toxiod BoNTa in ELISA. Recombinant ScFv-Fc had similar relative affinity to parent ScFv and had improved stability.