Improvement in affinity and thermostability of a fully human antibody against interleukin-17A by yeast-display technology and CDR grafting

• 来源期刊: 药学学报（英文版）
• 年，卷（期）: 2019,（5）
• 所属期刊栏目: Original articles
• 页码范围: 960-972
• 页数: 13页
• 语种: 英文

摘要

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 na(i)ve phage-displayed library,we isolated the anti-lL-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 vitro affinity 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 and in vivo analyses 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.