PRODUCTION AND PROPERTIES OF A BIFUNCTIONAL FUSION PROTEIN THAT MEDIATES ATTACHMENT OF VERO CELLS TO CELLULOSIC MATRICES

The sequence Arg-Gly-Asp (RGD) in extracellular matrix proteins such a s fibronectin, collagen, and laminin mediates cell attachment by inter acting with proteins of the integrin family of cell surface receptors. A gene fusion encoding the RGD-containing peptide, fused to the C-ter minus of a cellulose-binding domain (CBD/RGD), was expressed in Escher ichia coli. Cultures produced up to 50 mg of CBD/RGD per titer, most o f which was extracellular. it was purified from the culture supernatan t by affinity chromatography on cellulose. CBD/RGD promoted the attach ment of green monkey Vero cells to polystyrene and cellulose acetate. Attachment was inhibited by smalt synthetic peptides containing the RG D sequence. CBD/RGD was as effective as collagen in promoting the atta chment of Vero cells to Cellsnow(TM) microcarriers. (C) 1995 John Wile y and Sons, Inc.