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利用抗体噬菌体展示技术，克隆了血管内皮生长因子人基因工程单链抗体， 抗体表达量达菌体总蛋白的４５％．用层析柱复性技术同步完成抗体的纯化和复性．生 物活性实验表明，该抗体不仅特异结合人ＶＥＧＦ_１６５而且竞争抑制ＶＥＧＦ_１６５与其受体的 结合．为了提高单链抗体的亲和力，采用错配ＰＣＲ法，随机突变抗体重链可变区基因， 建立次级抗体突变库，并从中筛选出高亲和力突变株．研究了抗体突变株蛋白质三维 结构特点及其与亲和力的关系，这些结果不仅为肿瘤血管靶向治疗奠定了基础，而且 为抗体的高效表达及其亲和力体外成熟提供了参考模型．
Antibody phage display technology was used to clone human VEGF gene human scFv antibody, the expression level of antibody reached 45% of the total bacterial protein. Purification and renaturation of the antibody were performed simultaneously using a chromatography renaturation technique. Bioactivity experiments showed that the antibody not only specifically binds to human VEGF-165 but also competitively inhibits the binding of VEGF-165 to its receptor. In order to improve the affinity of single chain antibody, a mismatch PCR method was used to randomly mutate the heavy chain variable region genes of the antibody to establish a secondary antibody mutant library, and select high affinity mutant strains therefrom. The three-dimensional structural features and their relationship with the affinity of the antibody mutant strains were studied. These results not only laid the foundation for tumor vascular targeting therapy, but also provided a reference model for the high expression of the antibody and its affinity in vitro maturation.