Optimized methods for IL-17A refolding and anti-IL17A Fab production for co-crystallization with small molecules
Method Summary
An Escherichia coli system was used for IL-17A expression. Two-way optimizations of the refolding conditions were implemented by exploring various ratios of oxidized and reduced forms of the oxido-shuffling reagents and the refolding duration. Yield enhancement of anti-IL-17A Fab production was achieved after generating a stable HEK293-F cell line expressing the Fab fragment by G418 selection of transfected cells. The purified IL-17A/anti-IL-17A Fab complex was characterized by SDS-PAGE and western blotting. Single crystals of the quaternary complex, IL-17A/Fab/HAP (high-affinity peptide) with small-molecule compounds derived from our DNA encoded library could be obtained using a batch method.
Author: Meng X, Zhang L, Wei H et al.
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