Open AccessOptimized Methods for IL-17A Refolding and Anti-IL17A Fab Production for Co-crystallization With Small Molecules
Author(s) -
Xiaoyun Meng,
Lanjun Zhang,
Hong Wei,
Furong Li,
Lihua Hu,
Haiyun Ma,
Qian Liu,
Xiaoyu Li,
Zhongchuan Liu
Publication year - 2020
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2019-0170
Subject(s) - crystallization , yield (engineering) , chemistry , molecule , combinatorial chemistry , small molecule , peptide , drug discovery , interleukin 17 , computational biology , biochemistry , materials science , biology , organic chemistry , cytokine , immunology , metallurgy
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.
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