Releasable nonvolatile mass-label molecules
View Patent ↗Releasable tag reagents for use in the detection and analysis of target molecules, particular in mass spectrometric analyzes are provided. Also provided are methods of detection that employ releasable tag reagents.
1. A method for multiplexing the detection of a target molecule comprising:
(a) obtaining a plurality of probes, each comprising a reactive group, a release group and a mass label comprising a biopolymer;
(b) contacting the target molecule with the plurality of probes to produce probe:target molecule complexes, wherein the target molecule is attached to the reactive group of the probe;
(c) releasing the mass labels from the probe:target molecule complexes to produce released mass labels; and
(d) determining the mass of the released mass labels by mass spectrometry, wherein
each reactive group in a probe:target molecule complex is associated with a unique set of mass labels.
2. The method of claim 1 , wherein a plurality of target molecules is contacted with the plurality of probes.
3. The method of claim 1 , wherein the set of mass labels are attached to the same probe.
4. The method of claim 1 , wherein the set of mass labels are attached to different probes.
5. The method of claim 1 , wherein the target molecule is immobilized onto a solid support.
6. The method of claim 4 , wherein a plurality of target molecules are immobilized onto the solid support at spaced locations.
7. The method of claim 1 , wherein the target molecule is selected from the group consisting essentially of a polynucleotide, an antigen, a ligand, a polypeptide, a carbohydrate, and a lipid.
8. The method of claim 7 , wherein the target molecule comprises a polynucleotide.
9. The method of claim 7 , wherein the target molecule comprises a polypeptide.
10. The method of claim 1 , wherein the reactive group comprises a biomolecule capable of specific molecular recognition.
11. The method of claim 10 , wherein the reactive group comprises a polypeptide.
12. The method of claim 11 , wherein the polypeptide is selected from the group consisting essentially of an antibody, an enzyme, a receptor, a regulatory protein, a nucleic-acid binding protein, a hormone, or a protein product of a display methods.
13. The method of claim 12 , wherein the polypeptide comprises an antibody.
14. The method of claim 12 , wherein the polypeptide comprises a product of a phage display method or a bacterial display method.
15. The method of claim 10 , wherein the reactive group comprises a polynucleotide or an oligonucleotide.
16. The method of claim 1 , wherein the unique set of mass labels comprises a mass label that indicates the presence of a specified component within the reactive group.
17. The method of claim 16 , wherein the mass label indicates the presence of the specified component at a specified location within the reactive group.
18. The method of claim 17 , wherein a reactive group comprising n specified components is associated with a unique set of mass labels having n members.
19. The method of claim 18 , wherein n is from 1 to 1000.
20. The method of claim 17 , wherein a reactive group comprising n specified components is associated with a unique set of mass labels having y members wherein n is less than y!/[x!(y−x)!]; and wherein x comprises the number of mass labels used per reactive group.
21. The method of claim 17 , wherein the plurality of probes each comprise a known reactive group having a known set of mass labels.
22. The method of claim 21 , wherein the plurality of probes are prepared by combinatorial synthesis.
23. The method of claim 17 , wherein the plurality of target molecules comprise a known chemical structure.
24. The method of claim 16 , wherein the target molecule comprises an expressed gene product.
25. The method of claim 24 , wherein the expressed gene product is derived from a cloned mRNA.
26. The method of claim 16 , wherein the target molecule comprises a cloned genomic DNA.
27. The method of claim 1 , wherein the biopolymer comprises one or more monomer units, wherein each monomer unit is separately and independently selected from the group consisting of an amino acid, a nucleic acid, and a saccharide.