摘要
目的 基因构建与表达SZ 63单链抗体 (SZ 63scFv) ,降低单克隆抗体对人体的免疫源性。方法 运用基因工程手段通过一段小分子连接肽 (Gly4 Ser) 3将SZ 63抗体的重链可变区基因和轻链可变区基因连接 ,构建成pET2 2b 63scFv表达载体 ,并导入大肠杆菌BL2 1(DE3 ) plys中进行表达 ,同时进行表达特性的研究。 结果 pET2 2b 63scFv在BL2 1(DE3 ) plys中经IPTG诱导后 ,SZ 63scFv以包涵体形式存在 ,经复性处理后具有与纤维蛋白D 二聚体结合的活性 ,其表达量占菌体总蛋白的 18% ,SZ 63scFv具有 13mg/ g湿菌的表达量。 结论 成功地获得了SZ
Objective To reduce the imunogenicity and mdecular weight and obtain the recombinant protein of single-chain Fv fragment of antifibrin monoclonal antibody.Methods Amplified the cDNAs coding for hoary and light variable regions of the mouse Mab SZ-63,and attached to the oligonucleotide of the linker peptide 3 by means of recombinant DNA technique,then it was introduced into E.Coli strain BL21 for expression.Results The recombinant protein existed in the form of inclusion body in the total strain protein.The molecular weight of the recombinant protein was 33.0 KD and its expressed quantity was 13 mg/g wet strain and accounted for 18% of the total strain protein.Conclusion It was proved by Western blot that the recombinant protein SZ-63scFc had the ability of the activity of anti-human fibrin D-Dimer the same as the previous antibody SZ-63.
出处
《同济大学学报（医学版）》
CAS
2003年第3期218-221,共4页
Journal of Tongji University(Medical Science)
