Improvement in affinity and thermostability of a fully human antibody against interleukin-17A by yeast-display technology and CDR grafting
Improvement in affinity and thermostability of a fully human antibody against interleukin-17A by yeast-display technology and CDR grafting
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通过酵母展示技术和 CDR 移植提高全人源抗白细胞介素 17A 抗体的亲和力和热稳定性
DOI：
10.1016/j.apsb.2019.02.007
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发表时间：
2019-09-01
影响因子：
14.5
通讯作者：
Cui，Bing
中科院分区：
文献类型：
--
作者：
Sun，Wei;Yang，Zhaona;Cui，Bing
单克隆抗体（mAb）由于其高特异性和识别广泛抗原的能力而广泛应用于许多领域。 IL-17A 可以单独诱导快速炎症反应，也可以与其他促炎细胞因子协同诱导快速炎症反应。越来越多的证据表明，IL-17A 信号传导的治疗干预为自身免疫性疾病和癌症提供了一种有吸引力的治疗选择。在这里，我们提出了一种组合方法来优化新型抗 hIL-17A 抗体的亲和力和热稳定性。我们从大型噬菌体展示文库中分离出抗 IL-17A mAb 7H9，它可以中和重组人 IL-17A 的作用。然而，适度的中和效力和较差的热稳定性限制了其治疗应用。然后使用酵母展示的随机诱变文库进行体外亲和力优化来生成8D3。这导致了四个关键氨基酸的变化，并在基于细胞的中和测定中提供了大约 15 倍的效力增加。 8D3 的互补决定区 (CDR) 进一步嫁接到 huFv 4D5 的稳定框架上，以提高热稳定性。由此产生的混合抗体 9NT/S 比其原始抗体具有更优异的稳定性和亲和力。基于人纤维肉瘤细胞的测定和小鼠体内分析表明，抗 IL-17A 抗体 9NT/S 有效抑制 IL-17A 诱导的促炎细胞因子的分泌。因此，这种领先的抗 IL-17A mAb 可能被用作治疗 IL-17A 相关疾病的潜在同类最佳候选药物。
Monoclonal antibodies (mAbs) are widely used in many fields due to their high specificity and ability to recognize a broad range of antigens. IL-17A can induce a rapid inflammatory response both alone and synergistically with other proinflammatory cytokines. Accumulating evidence suggests that therapeutic intervention of IL-17A signaling offers an attractive treatment option for autoimmune diseases and cancer. Here, we present a combinatorial approach for optimizing the affinity and thermostability of a novel anti-hIL-17A antibody. From a large naïve phage-displayed library, we isolated the anti-IL-17A mAb 7H9 that can neutralize the effects of recombinant human IL-17A. However, the modest neutralization potency and poor thermostability limit its therapeutic applications.In vitroaffinity optimization was then used to generate 8D3 by using yeast-displayed random mutagenesis libraries. This resulted in four key amino acid changes and provided an approximately 15-fold potency increase in a cell-based neutralization assay. Complementarity-determining regions (CDRs) of 8D3 were further grafted onto the stable framework of the huFv 4D5 to improve thermostability. The resulting hybrid antibody 9NT/S has superior stabilization and affinities beyond its original antibody. Human fibrosarcoma cell-based assays andin vivoanalyses in mice indicated that the anti-IL-17A antibody 9NT/S efficiently inhibited the secretion of IL-17A-induced proinflammatory cytokines. Therefore, this lead anti-IL-17A mAb might be used as a potential best-in-class candidate for treating IL-17A related diseases.