title: In-house preparation of hydrogels for batch affinity purification of glutathione S-transferase tagged recombinant proteins.
abstract: BACKGROUND
  Many branches of biomedical research find use for pure recombinant proteins for direct application or to study other molecules and pathways. Glutathione affinity purification is commonly used to isolate and purify glutathione S-transferase (GST)-tagged fusion proteins from total cellular proteins in lysates. Although GST affinity materials are commercially available as glutathione immobilized on beaded agarose resins, few simple options for in-house production of those systems exist. Herein, we describe a novel method for the purification of GST-tagged recombinant proteins.
  
  
  RESULTS
  Glutathione was conjugated to low molecular weight poly(ethylene glycol) diacrylate (PEGDA) via thiol-ene "click" chemistry. With our in-house prepared PEGDA:glutathione (PEGDA:GSH) homogenates, we were able to purify a glutathione S-transferase (GST) green fluorescent protein (GFP) fusion protein (GST-GFP) from the soluble fraction of E. coli lysate. Further, microspheres were formed from the PEGDA:GSH hydrogels and improved protein binding to a level comparable to purchased GSH-agarose beads.
  
  
  CONCLUSIONS
  GSH containing polymers might find use as in-house methods of protein purification. They exhibited similar ability to purify GST tagged proteins as purchased GSH agarose beads.
authors.affiliation: Department of Biopharmaceutical Sciences, University of Illinois, Chicago, IL 60612-7231, USA.
authors.forename: Jason S
authors.initials: JS
authors.lastname: Buhrman
authors.forename: Jamie E
authors.initials: JE
authors.lastname: Rayahin
authors.forename: Melanie
authors.initials: M
authors.lastname: Köllmer
authors.forename: Richard A
authors.initials: RA
authors.lastname: Gemeinhart
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005978:Glutathione / Q000737:chemistry; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D020100:Hydrogels / Q000737:chemistry; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D020100:Hydrogels; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005982:Glutathione Transferase; D005978:Glutathione