Luciferase labelling method
US5837465A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Feb 28, 1997 |
| Grant date | Nov 17, 1998 |
| Priority date | — |
| Expiry date | Feb 28, 2017 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N33/535
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
Luciferase is conjugated to a chemical entity, particularly to a specific binding agent such as an antibody, antigen or a nucleic acid, and more particularly an antibody, by (a) mixing the luciferase with one or more of D-luciferin, magnesium ions and adenosine triphosphate and (b) performing a covalent coupling reaction between the luciferase and the binding reagent using a covalent coupling reagent where the amount of D-luciferin, magnesium ions and/or adenosine triphosphate is sufficient to protect the luciferase activity against inhibition by the covalent coupling reagent. Preferably, step (a) is carried out by mixing the luciferase with its substrates in solution and preferably both magnesium and adenosine triphosphate are present as magnesium adenosine triphosphate (Mg.sup.2+ ATP), optionally together with D-luciferin. Also disclosed is a labeled chemical entity comprising a chemical entity conjugated to active luciferase as formed by the method. Preferably, the chemical entity is a specific binding agent suitable for use in a specific binding assay, preferably being an antibody, antigen or nucleic acid. When the binding agent is a nucleic acid, it is preferably an oligonucle…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.