Method of purifying keratinocyte growth factor (KGF)
AssigneeThe United States of America as represented by the Department of Health and Human Services
InventorsJeffrey S. Rubin, Paul W. Finch, Stuart A. Aaronson
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Abstract
Discoveries are disclosed that show particular aspects of recombinant DNA technology can be used successfully to produce hitherto unknown human keratinocyte growth factor (KGF) protein free of other polypeptides. These proteins can be produced in various functional forms from spontaneously secreting cells or from DNA segments introduced into cells. These forms variously enable biochemical and functional studies of this novel protein as well as production of antibodies. Means are described for determining the level of expression of genes for the KGF protein, for example, by measuring mRNA levels in cells or by measuring antigen secreted in extracellular or body fluids.
Claims1
A method of purifying keratinocyte growth factor (KGF) comprising:
- (i) Contacting a first solution comprising KGF with heparin that is bound to an inert support, wherein said contacting occurs under conditions in which KGF binds to heparin to form a KGF:heparin complex;
- (ii) dissociating KGF from the KGF:heparin complex to form a second solution comprising KGF;
- (iii) fractionating the second solution by HPLC, molecular sieving, or ion-exchange chromatography;
- and
- (iv) testing the fractions of step (iii) for KGF biological activity, thereby determining which fractions contain purified KGF.
Dependencies are inferred from the claim text.