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目的 构建噬菌体抗体库及筛选抗人肝癌特异性噬菌体抗体。 方法 以ＲＴ－ＰＣＲ法从经Ｂｅｌ７４０４细胞免疫的ＢＡＬＢ／Ｃ小鼠脾淋巴细胞扩增免疫球蛋白的Ｆｄ 段及к链基因，克隆到表达载体ｐＣＯＭＢ３Ｈ－ＳＳ中并将抗体Ｆａｂ 段表达于单链噬菌体表面，构建噬菌体抗体库；以Ｂｅｌ７４０４细胞为抗原对噬菌体抗体库进行４轮“吸附－洗脱－扩增”的亲和筛选，挑取部分克隆检测与抗原的结合活性。 结果 建成容量为２×１０６ＣＦＵ 的噬菌体抗体库，在亲和筛选过程中，噬菌体收获率逐轮得到提高，第４轮为第一轮的２４５ 倍，含Ｆａｂ 基因的克隆比率也由２６％ 增到８３％ ，挑取的１０ 个克隆中有８ 个与Ｂｅｌ７４０４细胞有结合活性。 结论 噬菌体抗体库技术系高效筛选体系，为肿瘤单克隆抗体的制备提供了有效的途径
Objective To construct phage antibody library and screen anti-human liver cancer-specific phage antibody. Methods Fd and к chain genes of immunoglobulin were amplified from splenic lymphocytes of BALB / c mice immunized with Bel7404 by RT-PCR and cloned into the expression vector pCOMB3H-SS. The Fab fragment of the antibody was expressed in single-stranded Phage antibody library was constructed on the surface of phage. The phage antibody library was subjected to 4 rounds of “adsorption-eluting-amplification” affinity screening using Bel7404 cell antigen as an antigen, and some clones were picked out to detect the binding activity to the antigen. Results As a phage antibody library with a capacity of 2 × 106 CFU was constructed, the phage yield was improved round by round in the affinity screening process. Round 4 was 245 times of the first round and the rate of cloning with Fab gene was also increased from 26% 83% of the 10 clones selected, and 8 out of the 10 clones selected had Bel7404 cell binding activity. Conclusion The phage antibody library technology is an efficient screening system that provides an effective method for the preparation of monoclonal antibodies