Rapid fluorescence tagging of glycans and other biomolecules with enhanced MS signals
AssigneeWaters Technologies Corporation
InventorsDarryl W. Brousmiche, Ying-Qing Yu
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Abstract
Reagents comprising MS active, fluorescent molecules with an activate functionality for reaction with amines useful in rapid tagging of biomolecules such as N-glycans, proteins, peptides, and amino acids, and uses thereof are taught and described. These MS active, fluorescent molecules may have three functional components such as a tertiary amino group or other MS active atom, a highly fluorescent moiety, and a functional group that rapidly reacts with amines.
Claims8
A compound of Formula I
- wherein
- R1 is O═C═N— or
- R2 is independently selected from —H, —C1-C8 alkyl, —C1-C8 cycloalkyl, halo, dialkylamino, CH2-dialkylamino, aminocarbonyl, alkoxycarbonyl, or alkoxy, but not Cl or O═C═N—;
- and
- R3 and R4 are independently selected from —H, alkyl, alkyl amino, alkylsulfonic acid, alkyl phosphonic acid, wherein either R3 or R4 is alkylamino, alkyl phosphonic acid, or alkylsulfonic acid.
A compound of Formula II
- wherein
- m=0-9;
- n=0-9;
- R1 is O═C═N—, S═C═N—, or
- R2 is independently selected from methylene, substituted nitrogen, oxygen, carbonyl, amide, ester, sulfur, sulfoxide, or sulfone;
- R3 and R4 are independently selected from —H, alkyl, alkyl amino, alkylsulfonic acid, alkyl phosphonic acid, wherein either R3 or R4 is alkylamino, alkyl phosphonic acid, or alkylsulfonic acid;
- and
- R5 is independently selected from —H, —C1-C8 alkyl, —C1-C8 cycloalkyl, halo, dialkylamino, CH2-dialkylamino, aminocarbonyl, alkoxycarbonyl, or alkoxy, but not Cl when R1 is O═C═N—, and when R1 is S═C═N, R5 is H.
A compound of the structural formula
A method for rapid tagging of a glycan comprising the steps of:
- reacting a precursor glycan or precursor amino acid with the compound of Formula I
- wherein
- R1 is O═C═N—, S═C═N—, or
- R2 is independently selected from —H, —C1-C8 alkyl, —C1-C8 cycloalkyl, halo, CH2-dialkylamino, dialkylamino, aminocarbonyl, alkoxycarbonyl, or alkoxy;
- R3 and R4 are independently selected from —H, alkyl, alkyl amino, alkylsulfonic acid, alkyl phosphonic acid, wherein R3 or R4 is alkylamino, alkyl phosphonic acid, or alkylsulfonic acid, and wherein R3 and R4 together with the nitrogen to which they are attached may form an optionally substituted 5- to 8-membered saturated or partially unsaturated ring, and forming an MS active, fluorescent glycan derivative or an amino acid derivative.
A method for rapid tagging of a glycan comprising the steps of reacting a precursor glycan or precursor amino acid with the compound of Formula II
- wherein
- R1 is O═C═N—, S═C═N, or
- R2 is independently selected from methylene, substituted nitrogen, oxygen, carbonyl, amide, ester, sulfur, sulfoxide, or sulfone;
- R3 and R4 are independently selected from —H, alkyl, alkyl amino, alkylsulfonic acid, alkyl phosphonic acid, wherein R3 or R4 is alkylamino, alkyl phosphonic acid, or alkylsulfonic acid, and wherein R3 and R4 together with the nitrogen to which they are attached may form an optionally substituted 5- to 8-membered saturated or partially unsaturated ring;
- and
- R5 is independently selected from —H, —C1-C8 alkyl, —C1-C8 cycloalkyl, halo, CH2-dialkylamino, dialkylamino, aminocarbonyl, alkoxycarbonyl, or alkoxy, but not Cl or O═C═N—, and not when R1 is S═C═N,
- and forming an MS active, fluorescent glycan derivative or an amino acid derivative.
Dependencies are inferred from the claim text.