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目的构建抗人B细胞淋巴瘤单链抗体(ScFv)库。方法从人B细胞淋巴瘤细胞系Raji细胞免疫的BALB/c小鼠脾细胞中提取mRNA,RT-PCR扩增重、轻链可变区基因片断,连接成ScFv基因并扩增,将其克隆到噬菌体载体pHEN1中,构建成单链噬菌体抗体库。结果抗体库容量为3.4×106,约100%的噬菌体基因中有ScFv基因的插入。结论成功构建了抗人B细胞淋巴瘤噬菌体单链抗体库,方便进一步筛选抗人B细胞淋巴瘤抗体。
Objective To construct an anti-human B cell lymphoma single chain antibody (ScFv) library. Methods mRNA was extracted from BALB / c mouse spleen cells immunized with Raji cells of human B lymphoma cell line. The heavy and light chain variable region genes were amplified by RT-PCR and ligated into ScFv gene. The clones were cloned Into phage vector pHEN1 to construct a single-stranded phage antibody library. As a result, the antibody library capacity was 3.4 × 106, and about 100% of the phage genes contained the insertion of the ScFv gene. Conclusion The anti-human B cell lymphoma phage single chain antibody library was successfully constructed for further screening of anti-human B cell lymphoma antibody.