DETECTION OF ELEVATED BASIC FIBROBLAST GROWTH-FACTOR DURING EARLY HOURS OF IN-VITRO ANGIOGENESIS USING A FAST ELISA IMMUNOASSAY

Basic FGF (bFGF) is a growth factor that is thought to play an importa nt role in angiogenesis. Available assays that are used to detect bFGF are long and cumbersome. Here, we present a fast, easy and sensitive sandwich-type enzyme immunoassay for bFGF detection. Our method is a m odification of the method described by Watanabe et al (Biochem. Biophy s. Res. Commun. 1991;175, 229). Two monoclonal antibodies for antigen capture and one noncongugated polycolonal antibody for antigen detecti on are used instead of using three monoclonal antibodies with the cong ugation of one of them for detection. There is no change in the sensit ivity of the assay with average detection limit of 1 pg/well. Acidic f ibroblast growth factor does not interfere with the assay. Using this method, samples from conditioned media of capillary endothelial cell c ulture before and after angiogenesis were measured. Associated with de tection of start of tube formation, basic FGF was elevated at 8 hours from angiogenic stimulation and peaked at 48 hour (4 times control), s howing for the first time in an in vitro system that there is a transi ent increase in endogenous bFGF accompanying early steps of angiogenes is which in turn may be the trigger for new capillary formation. (C) 1 994 Academic Press, Inc.