Human catechol-o-methyltransferase (comt) assay
AssigneeThe University of Hong Kong
InventorsShu Leong Ho, Wing Lok Ho, David Boyer Ramsden
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Abstract
Disclosed is an assay (method) to quantify the amounts of catecholamine-O-methyltransferase (COMT) protein in samples, such as extracts from cell cultures, body fluids, tissues, and environmental samples. It uses novel agents (anti-NE, COMT-NE, or COMT-epitope-NE) in combination with two previously described agents (anti-COMT and COMT) in a competitive ELISA system to achieve this aim.
Claims20
An antibody to NE-epitope or to a variant of NE-epitope.
An isolated polypeptide comprising NE-epitope or a variant of NE-epitope that binds with antibody to NE-epitope.
An isolated polynucleotide comprising a nucleic acid sequence encoding the NE-epitope or a variant NE-epitope that binds with antibody to NE-epitope.
A method for qualitative or quantitative detection of catechol-O-methyltransferase (COMT) in a sample, comprising of;
- a) providing a coated surface with a pre-determined amount of anti-COMT antibody;
- b) contacting a sample with the coated surface, for binding COMT, if present in the sample, to the coated surface;
- c) contacting a pre-determined amount of detectably labeled standard recombinant COMT-NE conjugate (S-COMT-NE) with the coated surface, for binding to the coated surface;
- d) contacting an anti-NE antibody to the coated surface, wherein the anti-NE antibody binds to the NE epitope on the S-COMT-NE when the S-COMT-NE has previously bound to the anti-COMT antibody of the coated surface;
- e) contacting the coated surface with an enzyme-labeled antibody, wherein the enzyme-labeled antibody binds to the anti-NE antibody when the anti-NE antibody has previously bound to the S-COMT-NE that is bound to the anti-COMT antibody;
- f) contacting the coated surface with a chemical enzyme label indicator which indicates the presence of the enzyme labeled antibody bound to the coated surface;
- and
- g) determining the presence of COMT in the sample, or amount of COMT present in the sample, based on the presence or amount of enzyme labeled antibody bound to the coated surface.
Dependencies are inferred from the claim text.