Department of Biology, Utah State University, Logan, UT, 84322, USA.
Department of Biology, Utah State University, Logan, UT, 84322, USA; Department of Biological Engineering, Utah State University, Logan, UT, 84322, USA.
Protein Expr Purif. 2022 Nov;199:106152. doi: 10.1016/j.pep.2022.106152. Epub 2022 Aug 10.
The purpose of this study was to determine a method to purify recombinant hagfish intermediate filament proteins, alpha and gamma, in a scalable manner. The study succeeded by having an increase in protein recovery of up to 35% when comparing centrifuge purification and the developed tangential flow purification. The proteins were approximately the same purity of 70% pure but further purification increased the purity of the proteins by 16%, based on ImageJ analysis. The developed tangential flow filtration purification and final purification methods could be easily scaled up to meet industry scale purification needs. The scaled-up processes described in this study did not interfere with fiber production or formation, indicating the methods can produce usable proteins for material development.
本研究旨在确定一种可规模化纯化重组八目鳗中间丝蛋白α和γ的方法。通过比较离心纯化和开发的切向流纯化,该研究成功将蛋白回收率提高了 35%。基于 ImageJ 分析,两种蛋白的纯度大致相同,均为 70%,但进一步的纯化可将蛋白纯度提高 16%。开发的切向流过滤纯化和最终的纯化方法可轻松放大,以满足工业规模的纯化需求。本研究中描述的放大工艺不会干扰纤维的生产或形成,表明这些方法可生产用于材料开发的可用蛋白。