Optimized methods for IL-17A refolding and anti-IL17A Fab production for co-crystallization with small molecules

BioTechniques

Hong Wei¹  Zhongchuan Liu¹  Lanjun Zhang¹  Xiaoyu Li¹  Lihua Hu¹  Furong Li¹  Haiyun Ma¹  Qian Liu¹  Xiaoyun Meng¹ 

[1] 1Discovery Project Unit, HitGen Inc., Chengdu, China;

关键词: co-crystallization;    DNA encoded library;    Fab;    interleukin 17A (IL-17A);    oxido-shuffling reagent;    refolding;   

DOI  :  10.2144/btn-2019-0170

来源: DOAJ

【 摘 要 】

Refolding of human interleukin 17A (IL-17A) has been reported; however, the key refolding protocol was not robust enough to deliver consistent results and to be easily scaled up for crystallization. Here we report an optimized refolding method for IL-17A. Although co-crystal structures of IL-17A with ligands have been obtained with a high-affinity peptide and an anti-IL-17A Fab as stabilizers, neither the production yield nor the characterization of the IL-17A/Fab complex was reported. To facilitate co-crystallization of IL-17A with small-molecule compounds derived from our DNA encoded library, we also describe the method for yield enhancement of anti-IL-17A Fab production and characterize the IL-17A/Fab complex for the first time, providing an essential prerequisite for structure-based drug discovery targeting IL-17A.

【 授权许可】

Unknown