US 7,067,269 B2 · Assaying apparatus, kit, and method for lipids and associated enzymes

Abstract

A lipid assay method, kit, and apparatus involving exposure of a protein, having a lipid recognition motif that interacts with a target lipid and a competing lipid, to a solution containing the competing lipid, and determining whether the target lipid is present in the solution. The target lipid has a stronger affinity to the lipid recognition motif than does the competing lipid. The lipid recognition motif is preferably a pleckstrin homology (PH) domain, with the target lipid being a phosphoinositide. The assay determines activity of a lipid kinase, the target lipid being a phosphorylation product of a reaction between the lipid kinase and a substrate lipid. The assay can be a cancer screening method for detection of cancer cells, where detection of certain levels of a PI(3,4,5)P3 target lipid is an indicator of a cancer cell.

Claims10

Dependent claims

A method for the quantification of phosphoinositide kinase activity, which comprises:

exposing a test solution containing an unknown amount of a target lipid, wherein said target lipid is a phosphorylation product of a reaction between a phosphoinositide kinase that is added to said test solution and a substrate phosphoinositide lipid that is added to said test solution, to an analyte solution containing a protein having a phosphoinositide lipid recognition motif that interacts with said target lipid and a predetermined amount of a competing target lipid which is labeled by a non-radioactive signal, and measuring said signal wherein a decrease in signal correlates with an increase of the phosphoinositide kinase activity.

Dependencies are inferred from the claim text.

Facts

Application

09/991,933Filed 26 Nov 2001

Published

US 2003/0100028 A129 May 2003

Granted

27 Jun 2006

Term ends

Not estimatedThe source records don’t support an estimate.

Classification4 codes

G01N 33/575
Immunoassay; Biospecific binding assay; Materials therefor: for cancer
G01N 33/573
Immunoassay; Biospecific binding assay; Materials therefor: for enzymes or isoenzymes
G01N 33/92
Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing:…
G01N
Investigating or analysing materials by determining their chemical or physical properties

Assaying apparatus, kit, and method for lipids and associated enzymes

Life of this patent

Assaying apparatus, kit, and method for lipids and associated enzymes

Life of this patent

Abstract

Claims10