Improvement in affinity and thermostability of a fully human antibody against interleukin-17A by yeast-display technology and CDR grafting
| العنوان: | Improvement in affinity and thermostability of a fully human antibody against interleukin-17A by yeast-display technology and CDR grafting |
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| المؤلفون: | Zhao-na Yang, Bing Cui, Ming Liu, Wei Sun, Heng Lin, Chen-xi Zhao, Zhuo-Wei Hu, Xue-ying Hou |
| المصدر: | Acta Pharmaceutica Sinica. B Acta Pharmaceutica Sinica B, Vol 9, Iss 5, Pp 960-972 (2019) |
| بيانات النشر: | Elsevier BV, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | Monoclonal antibody, Original article, Phage display, medicine.drug_class, CDRs, complementarity-determining regions, Antibody engineering, LC, light chain, Yeast display, Neutralization, Proinflammatory cytokine, 03 medical and health sciences, 0302 clinical medicine, Antigen, FACS, fluorescent-activated cell sorting, AIN457, secukinumab, medicine, HRP, horse radish peroxidase, MFI, mean fluorescence intensity, General Pharmacology, Toxicology and Pharmaceutics, LY2439821, ixekizumab, Kon, the association rate constant, 030304 developmental biology, Thermostability, Koff, the dissociation rate constant, 0303 health sciences, biology, Chemistry, VH, the variable regions of heavy chains, lcsh:RM1-950, Antibody maturation, HC, heavy chain, scFv, single-chain variable fragment, YSD, yeast surface display, CDR grafting, KD, dissociation constant, lcsh:Therapeutics. Pharmacology, MACS, magnetic-activated cell sorting, Biochemistry, 030220 oncology & carcinogenesis, biology.protein, VL, the variable regions of light chains, Antibody, Yeast surface display, mAbs, monoclonal antibodies |
| الوصف: | 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 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. Graphical abstract Anti-IL-17A antibodies were initially developed from a naïve fully human antibody library. The candidate was further affinity-matured by constructing a library of yeast-displayed single-chain Fv (scFv) mutants and thermostability-improved by CDR grafting. The lead anti-IL-17A mAb 9NT/S might be used as a potential best-in-class candidate for treating IL-17A related diseases.fx1 |
| تدمد: | 2211-3835 |
| DOI: | 10.1016/j.apsb.2019.02.007 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3f1dc02e7d6958c545a29b9e66b7d4b7 https://doi.org/10.1016/j.apsb.2019.02.007 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....3f1dc02e7d6958c545a29b9e66b7d4b7 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 22113835 |
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| DOI: | 10.1016/j.apsb.2019.02.007 |