Original articles
Wei Sun, Zhaona Yang, Heng Lin, Ming Liu, Chenxi Zhao, Xueying Hou, Zhuowei Hu, Bing Cui. Improvement in affinity and thermostability of a fully human antibody against interleukin-17A by yeast-display technology and CDR grafting[J]. Acta Pharmaceutica Sinica B, 2019, 9(5): 960-972

Improvement in affinity and thermostability of a fully human antibody against interleukin-17A by yeast-display technology and CDR grafting
Wei Sun, Zhaona Yang, Heng Lin, Ming Liu, Chenxi Zhao, Xueying Hou, Zhuowei Hu, Bing Cui
State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China
Abstract:
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 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 cellbased 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.
Key words:    Monoclonal antibody    Antibody maturation    Phage display    Yeast surface display    CDR grafting    Antibody engineering   
Received: 2018-11-08     Revised: 2018-11-24
DOI: 10.1016/j.apsb.2019.02.007
Funds: This work was partially supported by National Key R&D Program of China under Grant 2017YFA0205400, National Natural Science Foundation of China under Grant 81874316, 81773781 and 81530093, National Drug Innovation Major Project of China under Grant 2018ZX09711001-003-001, Chinese Academy of Medical Sciences (CAMS, Beijing, China) Central Publicinterest Scientific Institution Basal Research Fund under 2017PT31046 and 2018RC350004, CAMS Innovation Found for Medical Sciences (2016-I2M-3-008 and 2016-I2M-1-007).
Corresponding author: Bing Cui     Email:cuibing@imm.ac.cn
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Wei Sun
Zhaona Yang
Heng Lin
Ming Liu
Chenxi Zhao
Xueying Hou
Zhuowei Hu
Bing Cui

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