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Reversed-phase high-performance liquid chromatography assay for recombinant acidic fibroblast growth factor in E.
P DePhillips1, B Buckland, K Gbewonyo
1Merck Research Labs, West Point, PA 19406.
Abstract:
A reversed-phase HPLC assay for acidic Fibroblast Growth Factor (aFGF) expressed in E. coli is described. The assay was developed on a Polymer Labs. PLRP-S macroporous poly(styrene-benzene) column. A sample preparation procedure was developed to permit quantitation of aFGF in crude samples, such as cell suspensions and cell lysates. The assay was linear over the range of 25-140 micrograms/ml with an accuracy of 5%. By using a POROS column containing a stationary phase with "through-pores" and a superficial mobile phase velocity of 1440 cm/h, the analysis could be performed in 3 min. The polymeric supports used for this assay were durable; periodic washing with sodium hydroxide maintained column performance for extended periods.