Fluorescent haloalkyl derivatives of reporter molecules well retained in cells
US5362628A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Mar 5, 1993 |
| Grant date | Nov 8, 1994 |
| Priority date | — |
| Expiry date | Mar 5, 2013 |
Classification
- Technology area (CPC Y)Emerging Cross-Sectional Technologies
- CPC primaryY10S530/802
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The subject invention provides a method for analyzing the metabolic activity in cells by improving the retention of a detectable reporter molecule only in intact cells where a particular enzyme is present. In particular, improved retention results from a two part process involving conjugation of haloalkyl-substituted derivatives of a reporter molecule with intracellular cysteine-containing peptides while unblocking the reporter molecule. The method for analyzing metabolic activity of cells involves the use of a substrate having the form EQU XR-REPORTER-BLOCK PA1 wherein -BLOCK is a group selected to be removable by action of a specific analyte, to give REPORTER spectral properties different from those of the substrate, PA1 -REPORTER- is a molecule that, when no longer bound to BLOCK by a BLOCK-REPORTER bond, has spectral properities different from those of the substrate, and PA1 XR-- is a haloalkyl moiety that can covalently react with an intracellular thiol (Z--S--H) to form a thioether conjugate (Z--S--R--). After the substrate enters the cells, the analyte removes BLOCK to make REPORTER detectable by the change in spectral properties, and the haloalkyl XR reacts with the intrace…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.