Expression and purification of bioactive, low-endotoxin recombinant human vitronectin. (June 2014)

• Record Type: Journal Article
• Authors: Halford, Michael M.; He, Yi-Chao; Stacker, Steven A.
• Is Part Of: Biotechniques. Volume 56:Number 6(2014)
• Page Start: 331
• Page End: 333
• Publication Date: 2014-06
• Subjects: recombinant -- human -- vitronectin -- integrin -- endotoxin -- expression -- purification -- endothelial; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000114181
• Languages: English
• ISSNs: 0736-6205

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.