Data on using single- and mixed-mode resins for capture chromatography of recombinant human thioredoxin from Escherichia coli.

ABSTRACT: This paper provides the data collected from screening chromatographic resins for their ability to bind and purify recombinant human thioredoxin from Escherichia coli lysate. This data was used by "Capture chromatography with mixed-mode resins: A case study with recombinant human thioredoxin from Escherichia coli" [1] to determine the optimal resin to use as a capture step to initiate downstream processing of thioredoxin. Five chromatography resins were screened using a 96-well filter plate to experiment on a wide range of pH and conductivity conditions in a shorter amount of time while saving on materials. Thioredoxin-producing E. coli was cultivated, harvested, and lysed according to Ravi et al [1]. Thioredoxin containing lysate was dialyzed into the binding conditions, pH from 5.0 to 9.0 and conductivity from 2.0 to 10.0 mS, applied to each resin and incubated with shaking for 0.5 h. Data gathered after the incubation period consisted of host cell protein and thioredoxin concentrations remaining in the supernatant, which was considered flowthrough for the remainder of this study. Samples containing high concentrations of thioredoxin after the experimental period indicate that thioredoxin did not bind to the resin at those conditions and should not be utilized as a capture step. Additionally, samples that contain low concentrations of host-cell proteins after the experimental period indicate large amounts of host-cell proteins bound to the resin. The corresponding conditions may not contribute to higher purity. Operating all screening experiments at small volumes allows for selecting optimal binding conditions while minimizing the burden on upfront biomass production.

SUBMITTER: Ravi A 

PROVIDER: S-EPMC7683221 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature