Affinity maturation of antiHER2 monoclonal antibody MIL5 using an epitope-specific synthetic phage library by computational design
Affinity maturation of antiHER2 monoclonal antibody MIL5 using an epitope-specific synthetic phage library by computational design
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DOI：
10.1080/07391102.2012.706073
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发表时间：
2013-04
影响因子：
4.4
通讯作者：
C. Qiao;M. Lv;Xinying Li;Jing Geng;Yan Li;Jiyan Zhang;Zhou Lin;Jiannan Feng;B. Shen
中科院分区：
文献类型：
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作者：
C. Qiao;M. Lv;Xinying Li;Jing Geng;Yan Li;Jiyan Zhang;Zhou Lin;Jiannan Feng;B. Shen
在许多情况下，亲和性的增加主要等同于生物功效的提高。目前，包括噬菌体文库在内的展示方法被广泛用于获得高亲和力的抗体。传统文库方法主要侧重于互补决定区诱变，需要大量的变异组合筛选和较大的文库容量来寻找亲和性较高的克隆。本研究在对抗原(HER2) -抗体（MIL5）复合体三维复合体结构建模的基础上，对接触面关键残基进行预测和鉴定，指导噬菌体文库的合成设计。构建由MIL5_scFv突变体组成的表位特异性位点定向突变噬菌体文库，从中筛选出亲和力较高的单链抗体（M5scFv_ph）。随后的实验结果表明，新型抗体M5scFv_ph保留了与亲本抗体MIL5_scFv重叠的表位，并且在体内对卵巢癌异种移植物具有相似的肿瘤生长抑制活性。
Increased affinities mainly equal to improved biological efficacy in many cases. By now, display methods including phage library are widely exploited to obtain higher affinity antibodies. Traditional library methods mainly focus on complementary determining region mutagenesis, in which extensive screening of variant combinations as well as large library capacity is required to find higher affinity clones. In this study, based on the modeling 3D complex structure of antigen (HER2)–antibody (MIL5) complex, the key residues of contact surface were predicted and identified to guide the synthetic phage library design. Then, epitope-specific site-directed mutagenesis phage library comprised of MIL5_scFv mutants was constructed, from which a higher affinity single chain antibody (M5scFv_ph) was screened out. Following experimental results showed that the novel antibody M5scFv_ph retained superimposed epitope to the parent antibody MIL5_scFv, and possessed similar tumor growth inhibitory activity in vivo on ovarian carcinoma xenografts.