Scalable purification of recombinant structural proteins, hagfish intermediate filament α and ɣ, from inclusion bodies for fiber formation

Protein Expr Purif. 2022 Nov:199:106152. doi: 10.1016/j.pep.2022.106152. Epub 2022 Aug 10.

Abstract

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.

Keywords: Hagfish; Inclusion body; Protein purification; Recombinant protein; Tangential flow filtration (TFF); rHIF.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Filtration / methods
  • Hagfishes* / metabolism
  • Inclusion Bodies / metabolism
  • Intermediate Filaments / metabolism
  • Recombinant Proteins / chemistry

Substances

  • Recombinant Proteins