US5707805A
Expired / Lapsed
US
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Assay for detecting keratinocyte growth factor (KGF) and its activity
Assignee: United States Department of Health and Human Services
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Filing: 1995-05-31
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Granted: 1998-01-13
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Expires: 2015-05-31
Abstract
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.
Inventors
- 👤 Jeffrey S. Rubin
- 👤 Paul W. Finch
- 👤 Stuart A. Aaronson
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-03 10:59
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12N; C12N15/00; C12N15/09; C12N15/11; A; A61; A61P; A61P17/00; A61P17/02; A61P43/00; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/50; C07K14/501; C07K16/00; C07K16/18; C07K16/22; C12N15/63; C12N15/70; C12N15/79; A61K; A61K38/00; C07K2317/00; C07K2317/70; C07K2317/73; C07K2319/00; C07K2319/01; C07K2319/02; C07K2319/70; C07K2319/74; C07K2319/75; C12Q2600/00; C12Q2600/158
Claims Analysis
Total Claims: 6
Claim 1
Independent Claim
143 words
A method for assaying Keratinocyte Growth Factor (KGF) activity comprising: (a) in a first reaction, determining the level of 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 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 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.
Claim 2
Independent Claim
143 words
A method for assaying Keratinocyte Growth Factor (KGF) activity comprising: (a) in a first reaction, determining the level of 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 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 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.
Claim 3
Dependent Claim
34 words
An assay for detecting human Keratinocyte Growth Factor (KGF) comprising contacting a test sample suspected of containing KGF with an antibody which specifically binds KGF and determining the amount of polypeptide antibody complex formed.
Claim 4
Dependent Claim
34 words
An assay for detecting human Keratinocyte Growth Factor (KGF) comprising contacting a test sample suspected of containing KGF with an antibody which specifically binds KGF and determining the amount of polypeptide antibody complex formed.
Claim 5
Independent Claim
89 words
A method for detecting Keratinocyte Growth Factor (KGF) in a test sample comprising: (a) determining the level of 3 H thymidine incorporation into DNA of keratinocytes after adding, in the presence of a KGF-neutralizing antibody, a test sample suspected of containing KGF to a culture of keratinocytes grown to confluence and maintained in a serum-free medium; (b) performing step (a) in the absence of a KGF-neutralizing antibody; wherein a decrease in 3 H thymidine incorporation in (a) relative to (b) indicates the presence of KGF in the test sample.
Claim 6
Independent Claim
89 words
A method for detecting Keratinocyte Growth Factor (KGF) in a test sample comprising: (a) determining the level of 3 H thymidine incorporation into DNA of keratinocytes after adding, in the presence of a KGF-neutralizing antibody, a test sample suspected of containing KGF to a culture of keratinocytes grown to confluence and maintained in a serum-free medium; (b) performing step (a) in the absence of a KGF-neutralizing antibody; wherein a decrease in 3 H thymidine incorporation in (a) relative to (b) indicates the presence of KGF in the test sample.
References Cited (Backward Citations)
WO1995001434A1
Cited
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