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 as fibronectin, collagen, and laminin mediates cell attachment by interacting with proteins of the integrin family of cell surface receptors. A gene fusion encoding the RGD-containing peptide, fused to the C-terminus of a cellulose-binding domain (CBD/RGD), was expressed in Escherichia coli. Cultures produced up to 50 mg of CBD/RGD per titer, most of which was extracellular. it was purified from the culture supernatant by affinity chromatography on cellulose. CBD/RGD promoted the attachment of green monkey Vero cells to polystyrene and cellulose acetate. Attachment was inhibited by smalt synthetic peptides containing the RGD sequence. CBD/RGD was as effective as collagen in promoting the attachment of Vero cells to Cellsnow(TM) microcarriers. (C) 1995 John Wiley and Sons, Inc.