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

BioTechniques. 2020-04-01;

Xiaoyun Meng, Lanjun Zhang, Hong Wei, Furong Li, Lihua Hu, Haiyun Ma, Qian Liu, Xiaoyu Li, Zhongchuan Liu

Products/Services Used	Details	Operation
Codon Optimization	… A cDNA encoding human IL-17A (20-155) without tag, was codon-optimized and synthesized by Genscript, then inserted into Nde I and Hind III sites of a pET30a plasmid (Novagen, Cat# 69909-3). The expression vector was transformed into E. coli BL21 (DE3), and isolation of …	Get A Quote

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 comple... More