Cloning, purification, and characterization of a non-collagenous anti-angiogenic protein domain from human α1 type IV collagen expressed in Sf9 cells

alpha 1(IV)NC1, a cleavage fragment of the carboxy terminal non-collagenous human alpha 1. chain of type IV collagen, is derived from the extracellular matrix specifically by MMP-2. Recently we determined the in vitro and in vivo anti-angiogenic activity of alpha 1(IV)NC1 and presently, its role in cancer therapy is under evaluation. To characterize alpha 1(IV)NC1 as a potential candidate for drug development and to test its efficacy in animal models, an effective method to produce a purified active form of alpha 1(IV)NC1 is needed. In the present study, expression of alpha 1(IV)NC1 in Sf9 cells using baculovirus expression system was discussed, this method was found to be effective in the production of a functionally active soluble form of the recombinant protein. The purified protein showed its characteristic activities such as inhibiting cell proliferation, migration, and tube formation in endothelial cells. (c) 2006 Elsevier Inc. All rights reserved.