Dual-labeled molecular constructs for detection of biochemical reactions
View Patent ↗The present invention relates to molecular constructs and methods of their use in detecting biochemical reactions. In particular, the invention relates to a molecular construct having a capture portion and a substrate portion, where the capture portion isolates the construct from a sample medium, and the substrate portion enables the construct to be acted upon and undergo a physical change which can be detected and measured. These molecular constructs may be used in diagnostic assays, genetic screening for potential risks of certain diseases in individuals, and drug discovery and toxicogenomics, using high throughput screening of compounds.
1. A method for identifying substrate peptides in a library of PNA-peptide chimeras, comprising:
providing a library of PNA-peptide chimeras, wherein the PNA sequence in the chimeras includes a subsequence which is complementary to a subsequence of a capture probe, and wherein said capture probe is bound to a solid support;
incubating the library of PNA-peptide chimeras with an enzyme of interest under conditions suitable for enzyme-substrate reactions to take place so as to transform the substrate peptides;
washing to remove the reaction products not attached to the solid supports;
determining, by examining the solid supports which substrates were transformed by the enzyme, thereby identifying substrate peptides.
2. The method of claim 1 , wherein the solid support is a bead.
3. The method of claim 2 , wherein the bead is encoded to provide the identification of the capture probe bound thereto.
4. The method of claim 3 , wherein the encoding is by fluorescence.