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目的：表达并纯化小鼠及人源化鼠抗人交联纤维蛋白单链抗体，并初步分析比较两者的体外生物活性。方法：经ＤＮＡ重组构建重组表达质粒，经ＩＰＴＧ诱导，在大肠杆菌中高表达两种单链抗体，用８ｍｏｌ／Ｌ尿素溶解包涵体后经ＩＭＡＣ纯化表达产物，并用凝血酶切除Ｎ端融合的（Ｈｉｓ）６，纯化的表达产物经复性后用ＥＬＩＳＡ检测其活性。结果：两种单链抗体在大肠杆菌中表达量占全菌蛋白的５０％以上，经变性、纯化后纯度可达９７％，而且两种复性后的纯化产物具有相似的抗原结合特性。结论：小鼠及人源化鼠抗人交联纤维蛋白单链抗体都能在大肠杆菌中实现高表达，而且两种重组单链抗体具有相似的抗原结合活性
OBJECTIVE: To express and purify mouse and humanized mouse anti-human cross-linked fibrin scFv antibodies and to analyze and compare the biological activity of the two in vitro. Methods: Recombinant plasmids were constructed by DNA recombination. Two kinds of single chain antibodies (McAbs) were expressed in Escherichia coli induced by IPTG. The inclusion bodies were dissolved in 8mol / L urea and then expressed by IMAC. The fusion protein of N His) 6, the purified expression product was refolded and its activity was detected by ELISA. Results: The two kinds of single chain antibodies expressed more than 50% of the total bacterial protein in E. coli. After purification, the purity of the two kinds of single chain antibodies reached 97%. And the purified products of the two refolded products had similar antigen-binding properties. CONCLUSION: Both mouse and humanized mouse anti-human cross-linked fibrin scFv can be highly expressed in E. coli, and both recombinant single chain antibodies have similar antigen binding activity