Resonance energy transfer assay with cleavage sequence and spacer
US8969016B2 · kind B2 · utility
Assignee
Inventors
Key dates
| Filing date | Sep 24, 2012 |
| Grant date | Mar 3, 2015 |
| Priority date | — |
| Expiry date | Sep 24, 2032 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/952
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
A molecular construct comprises a donor label, an acceptor label, a linker peptide disposed between the donor and the acceptor, the linker having a cleavage site sequence, and a spacer between at least one of (a) the donor and the cleavage site sequence and (b) the acceptor and the cleavage site sequence. Preferably, the construct is selected from the group consisting of CFP-(SGLRSRA)-SNAP-25-(SNS)-YFP, and CFP-(SGLRSRA)-synaptobrevin-(SNS)-YFP. In preferred embodiments, the linker peptide is a substrate of a botulinum neurotoxin selected from the group consisting of synaptobrevin (VAMP), syntaxin and SNAP-25, or a fragment thereof that can be recognized and cleaved by the botulinum neurotoxin. Advantageously, the spacer increases the electronic coupling between the donor label and the acceptor label relative to a corresponding construct without the spacer.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.