Optimized Methods for IL-17A Refolding and Anti-IL17A Fab Production for Co-crystallization With Small Molecules

Abstract

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.

Two Epitope Regions Revealed in the Complex of IL-17A and Anti-IL-17A VHH Domain

“…The procedure of inclusion bodies isolation, denaturation, and following refolding of IL-17A was similar with the protocol of Meng [ 34 ]. The cell pellet was suspended in a buffer A containing 50 mM Tris-HCl, pH 8.0, 2 mM Na 2 EDTA, centrifuged at 15,000× g for 15 min.…”