Methods for detecting a plurality of analytes by mass spectrometry

• US 6,649,351 B2
• Filed: 11/09/2001
• Issued: 11/18/2003
• Est. Priority Date: 04/30/1999
• Status: Expired due to Term

• Current Assignee: Virologic, Inc. (Laboratory Corporation Of America Holdings)
• Original Assignee: Aclara Biosciences, Inc (Laboratory Corporation Of America Holdings)
• Inventors: Hernandez, Vincent S., Matray, Tracy J., Hooper, Herbert, Singh, Sharat, Chenna, Ahmed
• Primary Examiner(s): Whisenant, Ethan
• Assistant Examiner(s): TUNG, JOYCE
• Application Number:
  US10/008,593
• Publication Number:
  US 20020150927A1
• Time in Patent Office: 739 Days
• Field of Search: 435/6, 435/287.2, 536/23.1, 536/24.3, 536/25.4, 536/26.6
• US Class Current: 435/6.11
• CPC Class Codes: C07H 19/06 Pyrimidine radicals
• CPC Class Codes: C07H 19/10 with the saccharide radical esterified by phosphoric or polyphosphoric acids
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2521/319 Exonuclease
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
• CPC Class Codes: C12Q 2565/627 being a mass spectrometer
• CPC Class Codes: C40B 20/08 Direct analysis of the library members per se by physical methods, e.g. spectroscopy
• CPC Class Codes: C40B 40/08 Libraries containing RNA or DNA which encodes proteins, e.g. gene libraries
• CPC Class Codes: C40B 50/16 involving encoding steps
• CPC Class Codes: C40B 70/00 Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes
• CPC Class Codes: G01N 33/561 Immunoelectrophoresis

Abstract

The invention provides a method for detecting a target analyte, by: (a) contacting one or more target analytes with a set of first and second binding reagents under conditions sufficient for binding of a target analyte with the first and second binding reagents, each of the first binding reagents containing a cleavage-inducing moiety and a target binding moiety, each of the second binding reagents containing a tagged probe having a mass modifier region attached to a target binding moiety by a cleavable linkage, the cleavable linkage being susceptible to cleavage when in proximity to an activated cleavage-inducing moiety; (b) activating the cleavage-inducing moiety to release a tag reporter, and (c) detecting a mass of the tag reporter, the mass uniquely corresponding to a known target analyte.

Claims

1. A method for detecting one or more target analytes, comprising:

(a) contacting one or more target analytes with a set of first and second binding reagents under conditions sufficient for binding of a target analyte with said first and second binding reagents, each of said first binding reagents comprising a cleavage-inducing moiety and a target binding moiety, each of said second binding reagents comprising a tagged probe having a mass modifier region attached to a target binding moiety by a cleavable linkage, the mass modifier region of each second binding reagent having a unique mass and said cleavable linkage being susceptible to cleavage when in proximity to said cleavage-inducing moiety;

(b) activating said cleavage-inducing moieties to cleave the cleavable linkages in proximity thereof so that mass modifier regions are released, and detecting the unique masses of the released mass modifier regions to determine each of the one or more target analytes.

2. The method of claim 1, wherein said mass is detected using a mass spectrometry method selected from the group consisting of quadrupole mass spectrometry, ion trap mass spectrometry, time-of-flight mass spectrometry and tandem mass spectrometry.

3. The method of claim 1, wherein said one or more target analytes further comprise a plurality of different target analytes.

4. The method of claim 1, wherein said one or more target analytes further comprise a binding partner of a specific binding pair.

5. The method of claim 1, wherein said one or more target analytes are selected from a group consisting of polypeptides, proteins, peptides, sugars, polysaccharides, nucleic acids, lipids, and small molecules.

6. The method of claim 1, wherein said first or second binding reagent further comprises a binding partner of a specific binding pair.

7. The method of claim 1, wherein said target binding moiety is selected from the group consisting of ligand, antiligand, receptor, antibody, biotin, avidin, strepavidin, protein A and polynucleotide, or a functional fragment thereof.

8. The method of claim 1, wherein said cleavage-inducing moiety further comprises a photosensitizer or a chemi-activated sensitizer.

9. The method of claim 1, wherein said cleavage-inducing moiety is a sensitizer capable of generating singlet oxygen.

10. The method of claim 8, wherein said cleavage-inducing moiety is a sensitizer selected from the group consisting of benzophenome, 9-thioxanthone, eosin, 9,10,-dibromoanthraene, methylene blue, metallo-porphyrins, chloroperoxidase and myeloperoxidase.

11. The method of claim 1, wherein said cleavage-inducing moiety further comprises two or more cleavage-inducing moieties.

12. The method of claim 1, wherein said mass modifier region further comprises two or more mass modifier regions.

13. The method of claim 1, wherein said cleavable linkage is susceptible to cleavage by singlet oxygen.

14. The method of claim 1, wherein said tagged probes further comprise a capture ligand having specific binding activity for a capture agent.

15. The method of claim 14, wherein said target binding moiety of the tagged probes further comprises a capture ligand having specific binding activity for a capture agent.

16. The method of claim 14, further comprising binding a set of tagged probes with a capture agent.

17. The method of claim 15, further comprising the step of separating one or more cleaved tagged probes from un-cleaved or partially cleaved tagged probes.

18. The method of claim 14, wherein said capture ligand further comprises biotin or an antigen.

19. The method of claim 14, wherein said capture agent is selected from the group consisting of avidin, streptavidin, an antibody, a receptor, or a functional fragment thereof, having specific binding affinity to the capture ligand.