Improvement in affinity and thermostability of a fully human antibody against interleukin-17A by yeast-display technology and CDR grafting.
10.1016/j.apsb.2019.02.007
- Author:
Wei SUN
1
;
Zhaona YANG
1
;
Heng LIN
1
;
Ming LIU
1
;
Chenxi ZHAO
1
;
Xueying HOU
1
;
Zhuowei HU
1
;
Bing CUI
1
Author Information
1. 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.
- Publication Type:Journal Article
- Keywords:
AIN457, secukinumab;
Antibody engineering;
Antibody maturation;
CDR grafting;
CDRs, complementarity-determining regions;
FACS, fluorescent-activated cell sorting;
HC, heavy chain;
HRP, horse radish peroxidase;
KD, dissociation constant;
Koff, the dissociation rate constant;
Kon, the association rate constant;
LC, light chain;
LY2439821, ixekizumab;
MACS, magnetic-activated cell sorting;
MFI, mean fluorescence intensity;
Monoclonal antibody;
Phage display;
VH, the variable regions of heavy chains;
VL, the variable regions of light chains;
YSD, yeast surface display;
Yeast surface display;
mAbs, monoclonal antibodies;
scFv, single-chain variable fragment
- From:
Acta Pharmaceutica Sinica B
2019;9(5):960-972
- CountryChina
- Language:English
-
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. 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 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.
