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(Gly4Ser)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pET22b-63scFv表达载体,并导入大肠杆菌BL21(DE3)plys中进行表达,同时进行表达特性的研究。结果:pET22b-63scFv在BL21(DE)3plys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-63scFv单链抗体并保留了与亲本抗体同一的抗体结合特性。 |
关键词: 单克隆抗体 单链抗体 人纤维蛋白 免疫源性 |
DOI: |
修订日期:2003-03-06 |
基金项目: |
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Construction and Expression of Single Chain FV Fragment of Monoclonal Antibody against Human Cross-linked Fibrin and Its Immunological Characterization |
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Abstract: |
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,an |
Key words: monoclonal antibody,single chain antibody,human fibrin |