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目的 构建和表达抗CD3 单链抗体 ,并对其构象进行模拟分析。方法 应用重叠延伸拼接法连接抗CD3 抗体的轻链可变区基因和重链可变区基因 ,构建抗CD3 单链抗体基因 (ScFv) ,并将其克隆于原核表达载体中表达。结果 拼接后的抗CD3 ScFv基因全长 732bp ,构象模拟分析表明其三维结构稳定 ,兼容性分数S >0 ,表达的目的蛋白约2 6 5kD。结论 抗CD3 ScFv基因的成功构建 ,为进一步研究抗CD3 /抗乙肝病毒的双特异性抗体奠定了基础。
Objective To construct and express anti-CD3 scFv and to constract its conformation. Methods The variable light chain variable region and heavy chain variable region genes of anti-CD3 antibody were linked by overlap extension method and the anti-CD3 single-chain antibody gene (ScFv) was constructed and expressed in prokaryotic expression vector. Results The spliced anti-CD3 ScFv gene was 732bp in length. The conformational analysis showed that the three-dimensional structure was stable, the compatibility score was more than 0, and the expressed protein was about 2.56 kD. Conclusion The successful construction of anti-CD3 ScFv gene lays the foundation for the further study of anti-CD3 / anti-HBV dual-specificity antibody.