Multiplexed detection with isotope-coded reporters

Abstract

Some aspects of this invention provide reagents and methods for the sensitive, quantitative and simultaneous detection of target analytes in complex biological samples by liquid chromatography tandem mass spectrometry (LC MS/MS). Some aspects of this invention provide affinity reagents encoded with mass reporters for the sensitive and quantitative translation of an analyte of interest into a mass tag. The reagents and methods provided herein have general utility in analyte detection and encoding, for example, in biomolecular profiling, molecular diagnostics, and biochemical encoding.

1 - 103 . (canceled) 
     
     
         104 . A composition comprising a plurality of isotope-labeled molecules wherein each isotope-labeled molecule of said plurality of isotope-labeled molecules comprises:
 a) a carrier, and   b) a mass tag conjugated to said carrier via a cleavable linker, wherein said cleavable linker is cleaved by a disease-associated protease.   
     
     
         105 . The composition of  claim 104 , wherein said mass tag comprises an isobaric mass tag. 
     
     
         106 . The composition of  claim 104 , wherein said mass tag comprises a peptide mass tag. 
     
     
         107 . The composition of  claim 104 , wherein said isotope-labeled molecules are configured for use in mass spectrometry (MS). 
     
     
         108 . The composition of  claim 107 , wherein said isotope-labeled molecules are configured for use in liquid-chromatography mass spectrometry (LC-MS). 
     
     
         109 . The composition of  claim 107 , wherein said isotope-labeled molecules are configured for use in tandem mass spectrometry (MS/MS). 
     
     
         110 . The composition of  claim 104 , wherein said disease-associated protease is selected from the group consisting of MMP2, MMP7, MMP9, MMP12, MMP14, tissue factor, factor Xa, cathepsin, and thrombin. 
     
     
         111 . The composition of  claim 104 , wherein each isotope-labeled molecules of said plurality of isotope-labeled molecules is the same. 
     
     
         112 . The composition of  claim 104 , wherein said plurality of isotope-labeled molecules comprises at least 6 different mass tag-conjugated cleavable linkers. 
     
     
         113 . The composition of  claim 104 , wherein said plurality of isotope-labeled molecules comprises at least 10 different mass tag-conjugated cleavable linkers. 
     
     
         114 . The composition of  claim 104 , wherein said plurality of isotope-labeled molecules comprises at least 20 different mass tag-conjugated cleavable linkers. 
     
     
         115 . The composition of  claim 104 , wherein said cleavage occurs in vivo. 
     
     
         116 . The composition of  claim 104 , wherein said cleavage occurs in vitro. 
     
     
         117 . The composition of  claim 104 , wherein said cleavage occurs ex vivo. 
     
     
         118 . The composition of  claim 104 , wherein said carrier comprises a particle. 
     
     
         119 . The composition of  claim 118 , wherein said particle comprises a microparticle or a nanoparticle. 
     
     
         120 . The composition of  claim 104 , wherein said carrier further comprises a binding agent. 
     
     
         121 . The composition of  claim 120 , wherein said binding agent is selected from a group consisting of an antibody, an antibody fragment, an aptamer, and an adnectin. 
     
     
         122 . The composition of  claim 104 , wherein said cleavable linker comprises a peptide. 
     
     
         123 . The composition of  claim 104 , wherein each isotope-labeled molecule of said plurality of isotope-labeled molecules further comprises a reactive chemical moiety.