REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ASSAY FOR RECOMBINANT ACIDIC FIBROBLAST GROWTH-FACTOR IN ESCHERICHIA-COLI CELL-SUSPENSIONS AND LYSATE SAMPLES

A reversed-phase HPLC assay for acidic Fibroblast Growth Factor (aFGF) expressed in E. coil is described. The assay was developed on a Polym er Labs. PLRP-S macroporous poly(styrene-benzene) column. A sample pre paration procedure was developed to permit quantitation of aFGF in cru de samples, such as cell suspensions and cell lysates. The assay was l inear over the range of 25-140 mu g/ml with an accuracy of 5%. By usin g a POROS column containing a stationary phase with ''through-pores'' and a superficial mobile phase velocity of 1440 cm/h, the analysis cou ld be performed in 3 min. The polymeric supports used for this assay w ere durable; periodic washing with sodium hydroxide maintained column performance for extended periods.