Production of Anti-Fibroblast Growth Factor Receptor Monoclonal Antibodies by In Vitro Immunization

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Abstract
Monoclonal antibodies against the high affinity tyrosine kinase Fibroblast Growth Factor (FGF) receptor may help define receptor epitopes inuolued in FGF binding and signal transduction which mediate coronary and tumor angiogenesis (the deuelopment of blood uessels). Monoclonal antibodies against the FGF receptor were made by in uitro immunization. Receptor was PRGE-purified and transblotted to nitrocellulose. The nitrocellulose was dissolued with acetonitrile, and the receptor used as antigen in an in uitro immunization system. Screening for FGF receptor positiue clones was done using membrane preparations from Coronary Uenular Endothelial Cells (CUEC) expressing the FGF receptor, both by Enzyme Linked Immunosorbent Assay (ELISR) and western blot. Culture supernatants of seueral clones tested positiue for IgM or IgG monoclonal antibodies against the FGF receptor. Antibodies were affinity purified. Ascites fluid was produced in Balb/c mice primed with Incomplete Freund's adjuuant (IFA). The monoclonal antibodies were also found to be suitable for receptor immunoprecipitation. Immunocytochemistry was done on sections from a uariety of species. Further characterization of these antibodies, as well as the production of the remainder of a panel of anti-FGF receptor monoclonal antibodies, is underway.