EXPRESSION AND CHARACTERIZATION OF HUMAN TISSUE INHIBITOR OF METALLOPROTEINASES-1 IN A BACULOVIRUS-INSECT CELL SYSTEM

The baculovirus expression system was used to overexpress human tissue inhibitor of metalloproteinases-1 (TIMP-1). Approximately 5 mg of rec ombinant TIMP-1 was produced per 10(9) Sf9 insect cells infected with the recombinant baculovirus. The optimum time point for the production of biologically active rTIMP-1 was 20 hours postinfection. TIMP-1 act ivity was demonstrated by a soluble collagenase inhibition assay and r everse zymography. The baculovirus system has the advantage over previ ously described methods for generating human rTIMP-1 in that it genera tes large amounts of a fully glycosylated and active protein. (C) 1994 Academic Press, Inc.