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Optimized Methods for IL-17A Refolding and anti-IL17A Fab Production for Co-Crystallization With Small Molecules 发布时间:2020-04-27

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.



BIOTECHNIQUESVOL. 69, NO. 1

https://doi.org/10.2144/btn-2019-0170

Link: https://www.future-science.com/doi/10.2144/btn-2019-0170


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