Isobaric labels for mass spectrometric analysis of peptides and method thereof

• US 7,799,576 B2
• Filed: 01/27/2004
• Issued: 09/21/2010
• Est. Priority Date: 01/30/2003
• Status: Active Grant

• Current Assignee: DH Technologies Pte. Ltd.
• Original Assignee: DH Technologies Development Pte. Ltd. (Danaher Corporation)
• Inventors: Bartlet-Jones, Michael, Pappin, Darryl J. C.
• Primary Examiner(s): Gakh; Yelena G
• Application Number:
  US10/765,458
• Publication Number:
  US 20040219686A1
• Time in Patent Office: 2,429 Days
• Field of Search: None
• US Class Current: 436/173
• 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, mixtures, 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 method for mass spectrometric analysis of isobarically labeled reactive peptides comprising:

a) providing two or more samples, each sample comprising one or more reactive peptide analytes,

b) reacting a first sample of the two or more samples with a first labeling reagent of a set of isobaric labeling reagents,

c) reacting a second sample of the two or more samples with a second labeling reagent of the set of isobaric labeling reagents, the second labeling reagent differing isobarically from the first labeling reagent, wherein steps b) and c) thereby produce two or more differentially labeled samples each comprising one or more isobarically labeled peptide analytes,

d) mixing together the two or more differentially labeled samples to form a sample mixture comprising two or more isobarically labeled peptide analytes of the formula;

2. A method for mass spectrometric analysis of isobarically labeled reactive peptides comprising:

a) providing two or more samples, each sample comprising one or more reactive peptide analytes,

b) reacting a first sample of the two or more samples with a first labeling reagent of a set of isobaric labeling reagents,

c) reacting a second sample of the two or more samples with a second labeling reagent of the set of isobaric labeling reagents, the second labeling reagent differing isobarically from the first labeling reagent, wherein steps b) and c) produce two or more differentially labeled samples each comprising one or more isobarically labeled peptide analytes,

d) mixing together the two or more differentially labeled samples to form a sample mixture comprising two or more isobarically labeled peptide analytes of the formula;

3. A method for mass spectrometric analysis of isobarically labeled reactive peptides comprising:

a) reacting providing two or more samples, each sample comprising one or more reactive peptide analytes,

b) reacting a first sample of the two or more samples with a first labeling reagent of a set of isobaric labeling reagents,

c) reacting a second sample of the two or more samples with a second labeling reagent of the set of isobaric labeling reagents, the second labeling reagent differing isobarically from the first labeling reagent, wherein steps b) and c) produce two or more differentially labeled samples each comprising one or more isobarically labeled peptide analytes

d) mixing together the two or more differentially labeled samples to form a sample mixture comprising two or more isobarically labeled peptide analytes of the formula;

4. A method for mass spectrometric analysis of isobarically labeled reactive peptides comprising:

a) providing two or more samples, each sample comprising one or more reactive peptide analytes,

b) reacting a first sample of the two or more samples with a first labeling reagent of a set of isobaric labeling reagents,

c) reacting a second sample of the two or more samples with a second labeling reagent of the set of isobaric labeling reagents, the second labeling reagent differing isobarically from the first labeling reagent, wherein steps b) and c) produce two or more differentially labeled samples each comprising one or more isobarically labeled peptide analytes,

d) mixing together the two or more differentially labeled samples to form a sample mixture comprising two or more isobarically labeled peptide analytes of the formula;

5. A mixture comprising at least two labeled peptide analytes, wherein each of the two labeled peptide analytes originates from a different sample combined to form the mixture and each comprises a different isobaric label from a same set of isobaric labels, each isobaric label comprising 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 peptide analyte is linked through the carbonyl carbon of the N-alkyl acetic acid moiety, wherein each different isobaric label comprises one or more heavy atom isotopes.

6. The mixture of claim 5, wherein each of the at least two labeled analytes in the mixture comprises the formula:

7. The mixture of claim 5, wherein the mixture comprises one two or more labeled analytes of the formula:

8. The mixture of claim 5, wherein the mixture comprises two or more labeled analytes of the formula:

9. The mixture of claim 5, wherein the mixture comprises two or more labeled analytes of the formula: