Mass labels

Abstract

A reactive mass label for labelling a biological molecule for detection by mass spectrometry includes the following structure: X-L-M-S-Re, wherein X is a mass marker moiety, L is a cleavable linker, M is a mass normalization moiety, and S is a mass series modifying group including the following group: wherein J is C═O, K is NH, and n is 2 or J and K are both CH 2 and n is 1, m is at least 1; and Re is a reactive functionality for attaching the reactive mass label to a biological molecule.

1 - 45 . (canceled) 
     
     
         46 . A reactive mass label for labelling a biological molecule for detection by mass spectrometry, the mass label comprising the structure:
   X-L-M-S-Re   wherein X is a mass marker moiety, L is a cleavable linker, M is a mass normalization moiety, and S is a mass series modifying group comprising the following group:   
       
         
           
           
               
               
           
         
         wherein J is C═O, K is NH, and n is 2, or J and K are both CH 2  and n is 1; m is at least 1; and 
         Re is a reactive functionality for attaching the reactive mass label to a biological molecule. 
       
     
     
         47 . The reactive mass label according to  claim 46 , wherein the reactive functionality Re comprises one of the following groups:
 (a)   
       
         
           
           
               
               
           
         
         
           wherein R 4  and R 5  may be the same or different and are each independently selected from the group consisting of H, a substituted or unsubstituted straight or branched C 1 -C 5  alkyl group, a substituted or unsubstituted aliphatic cyclic group, a substituted or unsubstituted aromatic group, and a substituted or unsubstituted heterocyclic group; 
         
         (b) 
       
       
         
           
           
               
               
           
         
         
           wherein each of R 6  to R 9  is independently selected from the group consisting of H, a substituted or unsubstituted straight or branched C 1 -C 5  alkyl group, a substituted or unsubstituted aliphatic cyclic group, a substituted or unsubstituted aromatic group and a substituted or unsubstituted heterocyclic group; 
         
         (c) 
       
       
         
           
           
               
               
           
         
         
           wherein halo is a halogen, 
         
         (d) 
       
       
         
           
           
               
               
           
         
         
           wherein B is —NH 2  or —(CH 2 ) n —ONH 2 , wherein n is from 1 to 6, 
         
         e) 
       
       
         
           
           
               
               
           
         
       
       and
 f) 
 
       
         
           
           
               
               
           
         
         
           wherein each R 2  is independently selected from the group consisting of H, a substituted or unsubstituted straight or branched C 1 -C 6  alkyl group, a substituted or unsubstituted aliphatic cyclic group, a substituted or unsubstituted aromatic group and a substituted or unsubstituted heterocyclic group. 
         
       
     
     
         48 . The reactive mass label according to  claim 46 , wherein L comprises an amide bond. 
     
     
         49 . The reactive mass label according to  claim 46 , wherein X is a mass marker moiety comprising the following group: 
       
         
           
           
               
               
           
         
         wherein the cyclic unit is aromatic or aliphatic and comprises from 0-3 double bonds independently between any two adjacent atoms; each Z is independently selected from the group consisting of N, N(R 1 ), C(R 1 ), CO, CO(R 1 ), C(R 1 ) 2 , O and S; X is either N, C or C(R 1 ); each R 1  is independently selected from the group consisting of H, a substituted or unsubstituted straight or branched C 1 -C 6  alkyl group, a substituted or unsubstituted aliphatic cyclic group, a substituted or unsubstituted aromatic group and a substituted or unsubstituted heterocyclic group; and y is an integer from 0-10. 
       
     
     
         50 . The reactive mass label according to  claim 46 , wherein the mass marker moiety comprises a structure selected from the group consisting of: 
       
         
           
           
               
               
           
         
       
     
     
         51 . The reactive mass label according to  claim 50 , wherein the mass marker moiety comprises the structure: 
       
         
           
           
               
               
           
         
       
     
     
         52 . The reactive mass label according to  claim 46 , wherein the reactive mass label comprises the structure: 
       
         
           
           
               
               
           
         
         wherein, J is C═O, K is NH, and n is 2, or J and K are both CH 2  and n is 1; and m is any positive integer including 0. 
       
     
     
         53 . The reactive mass label according to  claim 52 , wherein m is 0. 
     
     
         54 . The reactive mass label according to  claim 51 , wherein the reactive mass label comprises one of the following structures: 
       
         
           
           
               
               
           
         
         wherein n is at least 1. 
       
     
     
         55 . A set of two or more reactive mass labels, wherein each label in the set is as defined in  claim 46  and wherein each mass normalisation moiety ensures that the corresponding reactive mass label has a desired aggregate mass, and wherein the set comprises:
 (a) a group of reactive mass labels wherein the mass marker moiety of each reactive mass label has a common mass, each reactive mass label in the group having a unique aggregate mass; or 
 (b) a group of reactive mass labels wherein the mass marker moiety of each reactive mass label in the group has a mass different from that of all other mass marker moieties in that group, and each reactive mass label in the group has a common aggregate mass; 
 and wherein all of the reactive mass labels in the set are distinguishable from each other by mass spectroscopy. 
 
     
     
         56 . The set of reactive mass labels according to  claim 55 , wherein the mass marker moiety of each reactive mass label in the set has a common mass and each reactive mass label in the set has a unique aggregate mass. 
     
     
         57 . The set of reactive mass labels according to  claim 55 , wherein the mass marker moiety of each reactive mass label in the set has a unique mass and each reactive mass label in the set has a common aggregate mass. 
     
     
         58 . The set of reactive mass labels according to  claim 55 , wherein each reactive mass label in the set further comprises a mass adjuster moiety D to form the following structure:
   M(D) y -L-X(D) z      wherein M is the mass normalisation moiety, X is the mass marker moiety, D is the mass adjuster moiety, L is the cleavable linker, y and z are integers of 0 or greater, and y+z is an integer of 1 or greater.   
     
     
         59 . The set of reactive mass labels according to  claim 58 , wherein the mass adjuster moiety is selected from:
 (a) an isotopic substituent situated within one or more of the mass marker moiety and the mass normalisation moiety; and   (b) substituent atoms or groups attached to one or more of the mass marker moiety and the mass normalisation moiety.   
     
     
         60 . The set of reactive mass labels according to  claim 58 , wherein the mass adjuster moiety is selected from the group consisting of a halogen atom substituent, a methyl group substituent, and  2 H,  15 N,  13 C or  18 O isotopic substituents. 
     
     
         61 . The set of reactive mass labels according to  claim 55 , wherein each reactive mass label in the set comprises an isotopic mass adjuster moiety * to form the following structure:
   X (*)n -L-M (*)m      wherein X is the mass marker moiety, L is the cleavable linker, M is the mass normalisation moiety, and * is the isotopic mass adjuster moiety, and n and m are integers of 0 or greater such that either:   (a) the mass marker moiety of each reactive mass label in the set has a common mass and each reactive mass label in the set has a unique aggregate mass; or   (b) wherein the mass marker moiety of each reactive mass label in the set has a unique mass and each reactive mass label in the set has a common aggregate mass.   
     
     
         62 . The set of reactive mass labels according to  claim 55 , wherein the set comprises two or more reactive mass labels having a structure selected from the group consisting of: 
       
         
           
           
               
               
           
         
         wherein * represents that the oxygen is O 18 , carbon is O 13  or the nitrogen is N 15 , and wherein each reactive mass label in the set comprises one or more * such that either: 
         (a) the mass marker moiety of reactive mass label in the set has a common mass and each reactive mass label in the set has a unique aggregate mass; or 
         (b) the mass marker moiety of each reactive mass label in the set has a unique mass and each reactive label in the set has a common aggregate mass. 
       
     
     
         63 . An array of reactive mass labels, comprising two or more sets of reactive mass labels as set forth in  claim 55 , wherein the aggregate mass of each of the reactive mass labels of any one set in the array is different from the aggregate mass of each of the reactive mass labels of every other set in the array. 
     
     
         64 . The array of reactive mass labels according to  claim 63 , wherein the mass series modifying group of each reactive mass label in at least one set has a common mass, the mass series modifying group in each of the reactive mass labels of any one set having a different mass from the mass series modifying groups in each of the reactive mass labels of every other set in the array. 
     
     
         65 . The array of reactive mass labels according to  claim 64 , wherein the mass series modifying group of each of the reactive mass labels of any one set has a different mass from the mass series modifying groups in each of the reactive mass labels of every other set in the array due to the presence of a different number of isotopic mass adjuster moieties *. 
     
     
         66 . The array of mass labels according to  claim 65 , wherein the array of mass labels comprises:
 a) a first set of the reactive mass labels, wherein the mass series modifying group of each reactive mass label in the set has the following structure:   
       
         
           
           
               
               
           
         
         b) a second set of the reactive mass labels, the mass series modifying group of each reactive mass label in the set has the following structure: 
       
       
         
           
           
               
               
           
         
         c) a third set of the reactive mass labels, wherein the mass series modifying group of each reactive mass label in the set has the following structure: