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US 7,094,553 B2Granted
Antibodies specific for ungulate PrP
AssigneesThe Regents of the University of California, The Scripps Research Institute
InventorsStanley B. Prusiner, Jiri G. Safar, R. Anthony Williamson, Dennis R. Burton
Life of this patent
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Abstract
The present invention provides antibodies that specifically bind with a high degree of binding affinity to a native ungulate PrPC and/or a denatured ungulate PrPSc, but not to a native ungulate PrPSc. Preferred antibodies find native bovine PrPC and treated PrPSc but not native bovine PrPSc and can be used in an assay to determine if a sample is infected with infectious prions, i.e. PrPSc.
Claims24
A method of detecting ungulate PrPSc, comprising the steps of:
- providing a sample suspected of containing ungulate PrPSc and ungulate PrPC;
- dividing the sample into a first portion and a second portion;
- contacting the first portion with a labeled antibody which binds to native ungulate PrPC with a binding affinity of Ka of 108 l/mol or more wherein the antibody is further characterized by not binding to PrPC of a mammal other than a ungulate;
- treating the second portion in a manner in which causes any ungulate PrPSc to assume a confirmation which binds to the antibody which binds to native ungulate PrPC;
- contacting the treated second portion with the labeled antibody;
- determining the level of binding of labeled antibody to PrPC in the first portion;
- determining the level of binding of labeled antibody to treated PrP protein in the second portion;
- and
- comparing the level of binding of labeled antibody to PrP protein in the first portion with the level of binding to labeled antibody in the second portion and thereby determining whether the sample comprises ungulate PrPSc protein.
6 more elements
A method of detecting ungulate PrPSc, comprising the steps of:
- providing a sample suspected of containing ungulate PrPSc and ungulate PrPC;
- dividing the sample into a first portion and a second portion;
- contacting the first portion with a labeled antibody which binds to native ungulate PrPC with a binding affinity of Ka of 108 l/mol or more wherein the antibody is further characterized by not binding to PrPC of a mammal other than a ungulate;
- treating the second portion in a manner in which causes any ungulate PrPSc to assume a confirmation which binds to the antibody which binds to native ungulate PrPC;
- contacting the treated second portion with the labeled antibody;
- determining the level of binding of labeled antibody to PrPC in the first portion;
- determining the level of binding of labeled antibody to treated PrP protein in the second portion;
- adjusting the determined level of binding of labeled antibody to treated PrP protein in the second portion to compensate for increasing binding affinity of PrPC protein resulting from the treating;
- and
- comparing the level of binding of labeled antibody to PrP protein in the first portion with the level of binding to labeled antibody in the second portion and thereby determining whether the sample comprises ungulate PrPSc protein.
7 more elements
An assay, comprising:
- a support surface;
- and
- an antibody bound to the surface of the support, the antibody characterized by an ability to bind denatured ungulate PrPSc with a binding affinity of 107 l/mole or more wherein the antibody is further characterized by not binding to PrPC of a mammal other than a ungulate.
A method of detecting prions in an ungulate, comprising:
- extracting tissue from an animal;
- dividing the sample into a first portion and a second portion;
- contacting a the first portion with an antibody characterized by its ability to bind a native cow PrPC in situ wherein the antibody is further characterized by not binding to PrPC of a mammal other than a ungulate to determine the a first PrP level;
- treating a second portion to denature PrPSc present in the second portion;
- and
- contacting the denatured second sample of the tissue with the antibody to determine a second PrP level;
- wherein the presence of PrPSc in said sample is detected by subtracting the first level from the second level and considering an effect on the treating on the second PrP level.
4 more elements
Dependencies are inferred from the claim text.