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 liter, 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 small synthetic peptides containing the RGD sequence. CBD/RGD was as effective as collagen in promoting the attachment of Vero cells to Cellsnow™ microcarriers. © 1995 John Wiley & Sons, Inc.