Kits Pertaining to Analyte Determination

• US 20110045516A1
• Filed: 01/16/2007
• Published: 02/24/2011
• Est. Priority Date: 01/30/2003
• Status: Active Grant

• Current Assignee: DH Technologies Development Pte. Ltd. (Danaher Corporation)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Pappin, Darryl J.C., Bartlet-Jones, Michael
• Granted Patent: US 8,679,773 B2
• Time in Patent Office: Days
• US Class Current: 435/18
• 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. A kit comprising:

a) a set of two or more reagents suitable for the labeling of analytes, each reagent of the set comprising the formula;

RP-X-LK-Y-RG

or a salt thereof wherein;

i) RG is a reactive group that is a nucleophile or an electrophile and that is capable of reacting with one or more of the analytes of a sample;

ii) RP is a reporter moiety that comprises a fixed charge or that is ionizable, wherein the gross mass of each reporter is different for each reagent of the set;

iii) LK is a linker moiety that links the reactive group and the reporter group, wherein the mass of the linker compensates for the difference in gross mass between the reporters for the different labeling reagents of the set such that the aggregate gross mass of the reporter and linker combination is the same for each reagent of the set;

iv) X is a bond between an atom of the reporter and an atom of the linker;

v) Y is a bond between an atom of the linker and an atom of the reactive group, wherein, once the labeling reagent is reacted with the reactive analyte, bond Y links the linker to the analyte; and

vi) bonds X and Y fragment in at least a portion of the labeled analytes when subjected to dissociative energy levels; and

b) one or more reagents, containers, enzymes, buffers or instructions.

2. The kit of claim 1, wherein,

i) RG is a reactive group that is an electrophile and that is capable of reacting with one or more of the analytes of the sample;

ii) RP is a reporter moiety that comprises a fixed charge or that is ionizable, wherein the gross mass of each reporter is different for each reagent of the set;

iii) LK is a linker moiety that links the reactive group and the reporter group, wherein;

a) the mass of the linker compensates for the difference in gross mass between the reporters for the different labeling reagents of the set such that the aggregate gross mass of the reporter and linker combination is the same for each reagent of the set; and

b) the linker comprises at least one heavy atom isotope and has the formula;

3. The kit of claim 1 or 2, wherein the kit comprises a proteolytic enzyme.

4. The kit of claim 1 or 2, wherein the reactive group of each reagent of the set is an active ester.

5. The kit of claim 4, wherein the alcohol moiety of the active ester is a group of the formula:

6. The kit of claim 5, wherein the active ester is an N-hydroxysuccinimide ester.

7. The kit of claim 1 or 2, wherein the reporter is a substituted or unsubstituted morpholine, piperidine or piperazine.

8. The kit of claim 1 or 2, wherein the reporter comprises a carboxylic acid, sulfonic acid or phosphoric acid group.

9. The kit of claim 1, wherein the linker is a carbonyl or thiocarbonyl group.

10. The kit of claim 1 or 2, wherein each reagent of the set is independently linked to a solid support through a cleavable linker.

11. The kit of claim 1 or 2, wherein all reagents of the set are isomeric.

12. The kit of claim 1 or 2, wherein all reagents of the set are isobaric.

13. The kit of claim 2, wherein all reagents of the set comprise the formula:

14. The kit of claim 13, wherein the set comprises one or more of the following four reagents:

15. The kit of claim 13, wherein the set comprises one or more of the following four reagents:

16. The kit of claim 13, wherein the set comprises one or more of the following four support bound reagents:

17. The kit of claim 1, wherein all reagents of the set comprise the formula:

18. The kit of claim 17, wherein the set comprises one or more of the following four reagents:

19. The kit of claim 1 or 2, wherein each reagent of the set is isotopically enriched with one or more heavy atom isotopes.

20. The kit of claim 19, wherein each heavy atom isotope is present in at least 80% isotopic purity, at least 93% isotopic purity or at least 96% isotopic purity.

21. A kit comprising a compound and one or more reagents, containers, enzymes, buffers or instructions wherein, the compound is 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.

22. The kit of claim 21, wherein the kit comprises a proteolytic enzyme.

23. The kit of claim 21, wherein the kit comprises two or more of the compounds, wherein said two or more compounds are related as a set of isomeric and/or isobaric labeling reagents.

24. The kit of claim 23, wherein each labeling reagent is independently linked to a solid support through a cleavable linker.

25. The kit of claim 21, wherein each reagent of the set is isotopically enriched with one or more heavy atom isotopes.

26. The kit of claim 25, wherein each heavy atom isotope is present in at least 80% isotopic purity, at least 93% isotopic purity or at least 96% isotopic purity in the reagent.