QUANTITATIVE-DETERMINATION OF RECOMBINANT FIBROBLAST GROWTH-FACTOR RECEPTOR IN BACULOVIRUS-INFECTED INSECT-CELL CULTURES

Members of the fibroblast growth factor protein family are involved in several biologically important processes, including angiogenesis, wou nd healing, and tumor growth and metastasis. Interactions of basic fib roblast growth factor (bFGF) and its receptors are of considerable pha rmacological importance. Attempts were made to produce gram quantities of a soluble extracellular form of basic fibroblast growth factor rec eptor type 1 (bFGFR1) in order to study the energetics of its interact ion with bFGF The aim of the present study was to develop a method for monitoring changes in concentration of bFGFR1 during its production b y large-scale baculovirus-infected insect cell culture. A simple rever se-phase high-performance liquid chromatographic assay was developed f or direct determination of the soluble receptor secreted into insect c ell-culture media. The method permitted cell-culture samples containin g varying amounts of fetal calf serum and bFGFR1 (10-30 mg/L) to be an alyzed without prior purification. The assay was linear for added rece ptor in the range of 1-7 mu g.