Reversed-phase high-performance liquid chromatography assay for recombinant acidic fibroblast growth factor in E. coli cell suspensions and lysate samples.
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.
DePhillips
P
P
Merck Research Labs, West Point, PA 19406.
Buckland
B
B
Gbewonyo
K
K
Yamazaki
S
S
Sitrin
R
R
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D016220:Fibroblast Growth Factor 1 / Q000032:analysis* / Q000235:genetics; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet
D011994:Recombinant Proteins; D016220:Fibroblast Growth Factor 1