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目的 用基因工程抗体技术构建抗日本血吸虫膜蛋白特异的单链抗体。 方法 用抗体框架区的通用引物 PCR扩增抗日本血吸虫膜蛋白特异性单克隆抗体 NP11- 4的 VH 及 VL 基因 ,测序分析其核苷酸序列。用VH 和 VL 基因在 p THA90质粒载体中构建成单链抗体基因并诱导表达。 结果 扩增获得 NP11- 4的 VH 和 VL基因 ,经测序分析确定为新发现的抗体可变区序列 ,构建成排列顺序为 VH- linker- VL 的单链抗体基因 ,经诱导表达出与硫氧环蛋白融合的单链抗体 ,大小约为 36 .2 k Da,以包涵体形式存在。 结论 成功地构建和表达了抗日本血吸虫膜蛋白特异性单链抗体。
Objective To construct a single chain antibody specific for the membrane protein of Schistosoma japonicum by genetic engineering antibody technique. Methods The VH and VL genes of anti-Schistosoma japonicum membrane protein-specific monoclonal antibody NP11-4 were amplified by PCR using the universal primers of the antibody framework region. The nucleotide sequences were analyzed by sequencing. The VH and VL genes were constructed into the pTHA90 plasmid vector as single chain antibody genes and induced for expression. Results The VH and VL genes of NP11-4 were amplified and sequenced. The newly discovered antibody variable region sequences were constructed and sequenced as VHH-linker-VL single chain antibody genes. Cyclophilin-fused single chain antibodies, approximately 36.2 kDa in size, exist as inclusion bodies. Conclusion The McAb against Schistosoma japonicum membrane protein was successfully constructed and expressed.