title: Expression and purification of bioactive, low-endotoxin recombinant human vitronectin.
abstract: The secreted adhesive glycoprotein vitronectin (VTN) is a multifunctional component of plasma and the extracellular matrix. A high-yielding, inexpensive, low endotoxin source of bioactive recombinant human vitronectin (rhVTN) is highly desirable for in vitro use in diverse cell culture systems ranging from basic research settings to clinical-grade production of human cells. We describe modifications to a previously reported heparin-based affinity chromatography procedure that improve yield and achieve efficient removal of endotoxin from washed and urea-solubilized human VTN inclusion bodies following standard autoinduction of expression in Escherichia coli. This simple procedure makes accessible the low-cost expression and purification of large quantities of bioactive rhVTN using basic equipment and facilitates its use in a spectrum of endotoxin-sensitive applications.
authors.affiliation: Tumour Angiogenesis Program, The Peter MacCallum Cancer Centre, St Andrews Place, East Melbourne, Victoria, Australia.
authors.forename: Michael M
authors.initials: MM
authors.lastname: Halford
authors.affiliation: Tumour Angiogenesis Program, The Peter MacCallum Cancer Centre, St Andrews Place, East Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Yi-Chao
authors.initials: YC
authors.lastname: He
authors.affiliation: Tumour Angiogenesis Program, The Peter MacCallum Cancer Centre, St Andrews Place, East Melbourne, Victoria, Australia; Sir Peter MacCallum Department of Oncology, The University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Stacker
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods; D004731:Endotoxins / Q000302:isolation & purification*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics*; D015870:Gene Expression; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D019096:Vitronectin / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D004731:Endotoxins; D011994:Recombinant Proteins; D019096:Vitronectin
keywords: endothelial; endotoxin; expression; human; integrin; purification; recombinant; vitronectin