US 5,707,805 A · Assay for detecting keratinocyte growth factor (KGF) and its activity

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.

Claims3

Dependent claims

A method for assaying Keratinocyte Growth Factor (KGF) activity comprising:

(a) in a first reaction, determining the level of .sup.3 H thymidine incorporation into DNA after adding an aliquot of a test sample comprising isolated KGF to a culture of keratinocytes grown to confluence and maintained in a serum-free medium;
(b) in a second reaction, determining the level of .sup.3 H thymidine incorporation into DNA after adding another aliquot of the test sample of step (a) to a culture of fibroblasts grown to confluence and maintained in a serum free medium;
and
(c) assessing KGF activity by comparing the results of the first reaction with the results of the second reaction, wherein a fold stimulation of .sup.3 H thymidine incorporation into the DNA in the first reaction that is at least about 50 times greater than in the second reaction indicates KGF activity.

1 more element

Dependencies are inferred from the claim text.

Assay for detecting keratinocyte growth factor (KGF) and its activity

Life of this patent

Assay for detecting keratinocyte growth factor (KGF) and its activity

Life of this patent

Abstract

Claims3