USE OF MONOCLONAL-ANTIBODY TO DETECT BONE MORPHOGENETIC PROTEIN-4 (BMP-4)

A monoclonal antibody that reacts with murine and human bone morphogen etic protein-4 (BMP-4) has been developed using recombinant BMP-4 as a n immunogen. The antibody that bound most tightly to recombinant murin e (rm)BMP-4 was selected, subcloned, and characterized. The specificit y of the antibody was confirmed using Western blot analysis and enzyme -linked immunosorbent assay (ELISA). The antibody reacts with murine a nd human BMP-4 in both the reduced and nonreduced condition; however, this antibody shows cross-reactivity with neither human BMP-2 nor TGF- beta 1. Thus, the produced antibody could recognize the disulfide-link ed dimeric structure of bioactive BMP-4, regardless of the species. Im munocytochemical study using this antibody successfully shows the cyto solic localization of BMP-4 in osteoinductive cells; i.e., BFO and Sao s-2 in which the level of mRNA for BMP-4 was proved to be constitutive ly high by Northern blot analysis. In addition, the antibody could dem onstrate the presence of BMP-4 in developmental bone formation in the alveolar bone of rat embryo by immunohistochemistry. The antibody coul d be used for a more sensitive approach for quantitative analysis of B MP-4.