Search Thermo Fisher Scientific
Large scale FPLC purification of GST produces >95% purity of target protein
Biomass (170g) containing overexpressed GST was lysed with 1.7L of lysis buffer and then 0.75L of lysate was loaded onto equilibrated 200mL columns (50mm x 100mm) of Thermo Scientific Pierce Glutathione Superflow Agarose (left) or Glutathione Sepharose™ 4 Fast Flow (GE Healthcare, right) at a linear flow rate of 30cm/hour. Columns were washed with binding buffer until the UV280 reached baseline; then bound protein was eluted with elution buffer, and fractions containing purified GST were pooled. Load, flow-through, wash, and eluate fractions were separated by SDS-PAGE, stained with Imperial Protein Stain (Part No. 24615) and evaluated using myImageAnalysis Software (Part No. 62237) to determine purity. Total yield, recovery, and purity were nearly identical for both resins.
