Methods, compositions and kits pertaining to analyte determination

• US 20060105416A1
• Filed: 12/28/2005
• Published: 05/18/2006
• Est. Priority Date: 01/30/2003
• Status: Abandoned Application

• Current Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Pappin, Darryl J.C., Bartlet-Jones, Michael
• Application Number:
  US11/319,685
• Publication Number:
  US 20060105416A1
• Time in Patent Office: Days
• US Class Current: 435/23
• CPC Class Codes: A61K 51/04 Organic compounds
• CPC Class Codes: C07D 207/46 with hetero atoms directly attached to the ring nitrogen atom
• CPC Class Codes: C07D 401/12 linked by a chain containing hetero atoms as chain links
• CPC Class Codes: G01N 2458/15 Non-radioactive isotope labels, e.g. for detection by mass spectrometry
• CPC Class Codes: G01N 33/532 Production of labelled immunochemicals
• CPC Class Codes: G01N 33/6842 Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins
• CPC Class Codes: G01N 33/6848 Methods of protein analysis involving mass spectrometry
• CPC Class Codes: Y10T 436/142222 Hetero-O [e.g., ascorbic acid, etc.]
• CPC Class Codes: Y10T 436/145555 Hetero-N
• CPC Class Codes: Y10T 436/147777 Plural nitrogen in the same ring [e.g., barbituates, creatinine, etc.]
• CPC Class Codes: Y10T 436/24 Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry

Abstract

This invention pertains to methods, kits and/or compositions for the determination of analytes by mass analysis using unique labeling reagents or sets of unique labeling reagents. The labeling reagents can be isomeric or isobaric and can be used to produce mixtures suitable for multiplex analysis of the labeled analytes.

Claims

1. An active ester compound that is a 5, 6 or 7 membered heterocyclic ring comprising a ring nitrogen atom that is N-alkylated with a substituted or unsubstituted acetic acid moiety to which the alcohol moiety of the active ester is linked through the carbonyl carbon of the N-alkyl acetic acid moiety, wherein the compound is isotopically enriched with one or more heavy atom isotopes.

2. The compound of claim 1, wherein the compound is isotopically enriched with three or more heavy atom isotopes.

3. The compound of claim 1, wherein the heterocyclic ring is substituted with one or more substituents.

4. The compound of claim 3, wherein the one or more substituents are alkyl, alkoxy or aryl groups.

5. The compound of claim 4, wherein the one or more substituents are protected or unprotected amine groups, hydroxyl groups or thiol groups.

6. The compound of claim 1, wherein the heterocyclic ring is aliphatic.

7. The compound of claim 1, wherein the heterocyclic ring is aromatic.

8. The compound of claim 1, wherein the heterocyclic ring comprises one or more additional nitrogen, oxygen or sulfur atoms.

9. The compound of claim 1, wherein active ester is an N-hydroxysuccinimide ester.

10. The compound of claim 1, wherein the compound is a salt.

11. The compound of claim 1, wherein the compound is a mono-TFA salt, a mono-HCl salt, a bis-TFA salt or a bis-HCl salt.

12. The compound of claim 1, wherein each incorporated heavy atom isotope is present in at least 80 percent isotopic purity.

13. The compound of claim 1, wherein each incorporated heavy atom isotope is present in at least 93 percent isotopic purity.

14. The compound of claim 1, wherein each incorporated heavy atom isotope is present in at least 96 percent isotopic purity.

15. A kit comprising a compound as claimed in claim 1, further comprising one or more reagents, containers, enzymes, buffers or instructions.

16. The kit of claim 15, wherein the kit comprises a proteolytic enzyme.

17. The kit of claim 15, wherein the kit comprises two or more compounds as claimed in claim 1, wherein said two or more compounds are related as a set of isomers and/or isobaric labeling reagents.

18. The kit of claim 17, wherein each labeling reagent is independently linked to a solid support through a cleavable linker.

19. The kit of claim 15, wherein the compound has the formula:

20. A method comprising:

a) reacting an analyte with the compound as claimed in claim 1 to thereby produce a labeled analyte; and

b) mixing the labeled analyte with one or more differentially labeled analytes.

21. The method of claim 20, further comprising analyzing the mixture in a mass spectrometer.

22. The method of claim 20, wherein the analysis in a mass spectrometer involves MS/MS analysis.