title: A method for the prevention of thrombin-induced degradation of recombinant proteins.
abstract: A new strategy to prevent degradation of recombinant proteins caused by non-specific cleavage by thrombin is described. We demonstrate that degradation due to non-specific cleavage of recombinant protein mediated by thrombin can be completely prevented by separation of thrombin from the recombinant protein on spin columns packed with heparin-sepharose. This method is generally applicable to all recombinant proteins that require the thrombin for the cleavage of affinity tags for purification. To our knowledge, this is the first report of an efficient and reliable method for the separation of residual thrombin from purified recombinant proteins.
authors.affiliation: Department of Chemistry & Biochemistry, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Dakshinamurthy
authors.initials: D
authors.lastname: Rajalingam
authors.forename: Karuppanan Muthusamy
authors.initials: KM
authors.lastname: Kathir
authors.forename: Koteshwara
authors.initials: K
authors.lastname: Ananthamurthy
authors.forename: Paul D
authors.initials: PD
authors.lastname: Adams
authors.forename: Thallapuranam Krishnaswamy Suresh
authors.initials: TK
authors.lastname: Kumar
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006493:Heparin / Q000737:chemistry; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012685:Sepharose / Q000737:chemistry; D013917:Thrombin / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D006493:Heparin; D012685:Sepharose; D013917:Thrombin