Isolation of fibroblast growth factor
AssigneeThe Salk Institute for Biological Studies
InventorsDenis Gospodarowicz, Andrew Baird, Peter Bohlen
Life of this patent
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Abstract
Basic Fibroblast Growth Factor (FGF) is substantially purified by the employment of affinity chromatography using heparin-linked support material. Described is a simplified three step procedure for extracting basic FGF from either mammalian brain or mammalian pituitary tissue. Salt precipitation, e.g., with ammonium sulfate is used to provide a partially purified precipitate that is then subjected to ion-exchange chromatography, e.g., using a Carboxymethyl-Sephadex column. Substantially pure basic FGF fractions are then obtained by fractionating the further partially purified fractions using affinity chromatography on a heparin-linked support e.g., Heparin-Sepharose.
Claims13
A method of obtaining substantially pure basic fibroblast growth factor comprising providing a biological sample containing significant levels of basic fibroblast growth factor, extracting the solubilized component of said biological sample, partially purifying basic FGF from said extract by a salt-precipitation step, then fractionating said partially-purified basic FGF-containing material so as to remove neutral and acidic proteins, and then treating said fractionated basic FGF-containing material using affinity chromatography employing heparin moieties linked to an insoluble support as the stationary phase and employing a salt gradient mobile phase of increasing salt concentration, and collecting liquid fractions containing substantially purified basic fibroblast growth factor therefrom.
A method of obtaining substantially pure basic fibroblast growth factor comprising providing material containing significant levels of basic fibroblast growth factor, and treating fractionated basic FGF-containing material using affinity chromatography employing heparin moieties bound to an insoluble support as the stationary phase and employing a salt gradient mobile phase of increasing salt concentration, and collecting liquid fractions containing substantially purified basic fibroblast growth factor therefrom.
A method of obtaining substantially pure basic fibroblast growth factor from an impure mixture containing significant levels of basic fibroblast growth factor, which method comprises applying said mixture to an affinity chromatography column comprising heparin moieties bound to a particulate support as the stationary phase, and eluting said basic FGF from said support with an aqueous eluant mobile phase of generally neutral pH and a salt concentration which increases and is above 0.6 M and collecting substantially purified basic fibroblast growth factor therefrom.
Dependencies are inferred from the claim text.