Mass labels

• US 7,294,456 B2
• Filed: 03/14/2001
• Issued: 11/13/2007
• Est. Priority Date: 03/14/2000
• Status: Expired due to Term

• Current Assignee: Electrophoretics Limited (Proteome Sciences, PLC)
• Original Assignee: Electrophoretics Limited (Proteome Sciences, PLC)
• Inventors: Schmidt, Gunter, Johnstone, Robert Alexander Walker, Thompson, Andrew Hugin
• Primary Examiner(s): Schultz; J. Douglas
• Assistant Examiner(s): Lundgren; Jeffrey S
• Application Number:
  US10/221,666
• Publication Number:
  US 20030194717A1
• Time in Patent Office: 2,435 Days
• Field of Search: None
• US Class Current: 435/4
• 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: C07K 1/1077 by covalent attachment of residues other than amino acids or peptide residues, e.g. sugars, polyols, fatty acids
• CPC Class Codes: C12Q 1/6872 involving mass spectrometry
• CPC Class Codes: C40B 40/00 Libraries per se, e.g. arrays, mixtures
• CPC Class Codes: G01N 2458/15 Non-radioactive isotope labels, e.g. for detection by mass spectrometry
• CPC Class Codes: G01N 33/6803 General methods of protein analysis not limited to specific proteins or families of proteins
• CPC Class Codes: Y10T 436/13 Tracers or tags

Abstract

Provided herein are sets of mass labels. Each mass label in a set includes 1) a mass marker moiety; 2) a mass normalisation moiety; and 3) a cleavable linker connecting the mass marker moiety to the mass normalisation moiety. Each mass marker moiety is characterised as having a mass different from that of all other mass marker moieties in the set as determined by mass spectrometry. Further, each mass normalisation moiety ensures that each mass label in the set has substantially the same mass as determined by mass spectrometry.

Claims

1. A set of two or more mass labels, each mass label in the set comprising a mass marker moiety attached via a cleavable linker to a mass normalisation moiety, the mass marker moiety being fragmentation resistant, wherein each mass normalisation moiety ensures that each mass label in the set has the same aggregate mass as determined by mass spectrometry, and wherein each mass marker moiety has a mass different from that of all other mass marker moieties as determined by mass spectrometry, and wherein all the mass labels in the set are distinguishable from each other by mass spectrometry.

2. A set of mass labels according to claim 1, in which each mass marker moiety in the set has the same structural core, and each mass normalisation moiety in the set has the same structural core that may be the same or different from the structural core of the mass marker moieties, and wherein each mass label in the set comprises one or more mass adjuster moieties, the mass adjuster moieties being attached to or situated within the structural core of the mass marker moiety and/or the structural core of the mass normalisation moiety, such that every mass marker moiety in the set comprises a different number of mass adjuster moieties and every mass label in the set has the same number of mass adjuster moieties.

3. A set of mass labels according to claim 2, each mass label in the set having the following structure:

M(A)_y-L-X(A)_z

wherein M is a mass normalisation moiety, X is a mass marker moiety, A is a mass adjuster moiety, L is a cleavable linker, y and z are integers of 0 or greater, and y+z is an integer of 1 or greater.

4. A set of mass labels according to claim 2 or claim 3, wherein the mass adjuster moiety is selected from:

(a) an isotopic substituent situated within the structural core of the mass marker moiety and/or within the structural core of the mass normalisation moiety, and (b) substituent atoms or groups attached to the structural core of the mass marker moiety and/or attached to the structural core of the mass normalisation moiety.

5. A set of mass labels according to claim 4, wherein the mass adjuster moiety is selected from a halogen atom substituent, a methyl group and ²H or ¹³C isotopic substituents.

6. A set of mass labels according to claim 5, wherein the mass adjuster moiety is a fluorine atom substituent.

7. A set of mass labels according to claim 1, wherein the cleavable linker attaching the mass marker moiety to the mass normalisation moiety is a linker cleavable by collision.

8. A set of mass labels according to claim 7, wherein the cleavable linker comprises an amide bond.

9. A set of mass labels according to claim 1, wherein the the mass normalisation moiety comprises a fragmentation resistant group.

10. A set of mass labels according to claim 9, wherein the mass normalisation moiety comprises a phenyl group.

11. A set of mass labels according to claim 1, wherein the mass marker moiety comprises a pre-ionised group.

12. A set of mass labels according to claim 11, wherein the mass marker moiety comprises an N-methyl pyridyl group, or a group selected from the following groups:

—

NH₂, —

NR₂, —

NR₃⁺, —

SR₃⁺, —

SO₃⁻

, —

PO₄⁻

, —

PO₃⁻

, —

CO₂⁻

—

,

13. A set of mass labels according to claim 3, wherein each of the labels in the set has the following structure:

14. A set of mass labels according to claim 13, wherein R is H, L is an amide bond, p=0, and A is an F atom.

15. An array comprising two or more sets of mass labels as defined in claim 1, wherein the aggregate mass of each of the mass labels of any one set in the array is different from the aggregate mass of each of the mass labels of every other set in the array.

16. An array of mass labels according to claim 15, wherein each mass label in at least one set comprises a mass series modifying group of the same mass as determined by mass spectrometry, the mass series modifying group in each of the mass labels of any one set having a different mass from the mass series modifying groups in each of the mass labels of every other set in the array.

17. An array of mass labels according to claim 16, wherein the mass series modifying groups are attached to the mass labels such that, upon cleaving the cleavable linker of the mass labels, the mass series modifying groups become detached from the mass marker moieties.

18. An array of mass labels according to claim 16 or claim 17, wherein the mass series modifying groups of each set in the array have the same structural core, and each mass label of any one set in the array has the same number of mass series modifying groups as the other mass labels of that set, and a different number of mass series modifying groups from the mass labels of every other set in the array.

19. An array of mass labels according to claim 18, each mass label in the array having either of the following structures:

(S)_x-M(A)_y-L-X(A)_z

M(A)_y-(S)_x-L-X(A)_z

wherein S is a mass series modifying group;

M is a mass normalisation moiety;

X is a mass marker moiety;

A is a mass adjuster moiety;

L is a cleavable linker;

x is an integer of 0 or greater;

y and z are integers of 0 or greater; and

y+z is an integer of 1 or greater.

20. An array of mass labels according to claim 16, wherein the mass series modifying groups comprise an aryl ether group.

21. An array of mass labels according to claim 19 or claim 20, each mass label in the array having either of the following structures:

22. An array of mass labels according to claim 16 or claim 19, wherein the mass series modifying groups of every set in the array have the same structural core, the mass series modifying group of the mass labels of at least one set comprising one or more mass adjuster moieties, the mass adjuster moieties being attached to or situated within the structural core of the mass series modifying group.

23. An array of mass labels according to claim 22, in which every mass label of every set in the array has the same number of mass series modifying groups, wherein the mass series modifying group in each mass label of any one set has the same number of mass adjuster moieties as the mass series modifying groups in every other label of that set, and wherein the mass series modifying groups in the mass labels of any one set have a different number of mass adjuster moieties from the mass series modifying groups in the labels of every other set in the array.

24. An array of mass labels according to claim 23, wherein each of the sets in the array comprises mass labels having either of the following structures:

S(A*)_r-M(A)_y-L-X(A)_z

M(A)_y-S(A*)_r-L-X(A)_z

wherein S is a mass series modifying group;

M is a mass normalisation moiety;

X is a mass marker moiety;

A is a mass adjuster moiety of the mass marker moieties and mass normalisation moieties;

A* may be the same or different from A and is a mass adjuster moiety of the mass series modifying groups;

L is a cleavable linker;

r is an integer of 0 or greater and is at least 1 for one or more sets of mass labels in the array;

y and z are integers of 0 or greater; and

y+z is an integer of 1 or greater.

25. An array of mass labels according to claim 24, wherein each of the sets in the array comprises mass labels having either of the following structures: