Discovery Project Unit, HitGen Inc., Chengdu, China.
Biotechniques. 2020 Jul;69(1):427-433. doi: 10.2144/btn-2019-0170. Epub 2020 Apr 27.
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.
已有人报道人白细胞介素 17A（IL-17A）的复性；然而，关键的复性方案不够稳健，无法获得一致的结果，也难以进行大规模放大以用于结晶。在这里，我们报告了一种用于 IL-17A 的优化复性方法。尽管已经获得了与配体结合的 IL-17A 的共晶结构，其中高亲和力肽和抗 IL-17A Fab 作为稳定剂，但均未报道 IL-17A/Fab 复合物的产率或特性。为了促进小分子化合物与我们的 DNA 编码文库衍生的小分子化合物共结晶，我们还描述了提高抗 IL-17A Fab 产量的方法，并首次对 IL-17A/Fab 复合物进行了表征，为靶向 IL-17A 的基于结构的药物发现提供了必要的前提条件。
