Method for the detection and measurement of Hapten-conjugated biological binding entities by western and dot-blot using anti-hapten antibodies

US7582443B2 · kind B2 · utility

Inventor

• Philip Cavanaugh · Houston, US

Key dates

• Filing date: Dec 5, 2001
• Grant date: Sep 1, 2009
• Expiry date: Dec 5, 2021

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10T436/10
• WIPO field: Measurement
• WIPO sector: Instruments

Abstract

The invention is a procedure for measuring the binding of an entity (ligand) to a surface by using a hapten-conjugated version of the ligand (hapten-ligand). An excess of the hapten-ligand is presented to the binding surface and excess (unbound) hapten-ligand is washed off. Bound hapten-ligand is then solubilized (removed) and applied to a membrane support or separated by electrophoresis and applied to a membrane support. Known amounts of hapten-ligand are similarly applied to the membrane, to provide for hapten-ligand standards. The membrane-bound hapten-ligand is detected by application of an enzyme-conjugated antibody to the hapten; or by application of an antibody to the hapten followed by application of an enzyme-conjugated antibody to the anti-hapten antibody. The resultant membrane-associated enzyme is detected and quantitated by the application of a color or light-producing substrate which reacts with the enzyme. A combination of the use of anti-hapten antibodies along with membrane-blotting technologies to assess hapten-ligand binding to surfaces is not found in the scientific or patent literature, particularly in regards to assessing protein binding to cell surfaces.