Sets and compositions pertaining to analyte determination
First Claim
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1. A set of two or more reagent compounds, or a salt thereof, each of the formula RP-X-LK-Y-RG, wherein:
- i) RG is a reactive group that is a nucleophile or an electrophile and that is capable of reacting with a functional group of one or more reactive analytes of a sample;
ii) RP is a reporter moiety that comprises a fixed charge or that is ionizable, wherein the gross mass of each reporter moiety is different for each reagent compound of the set;
iii) LK is a linker moiety that links the reactive group to the reporter moiety, wherein the mass of the linker compensates for the difference in gross mass between the reporter moieties for the different reagent compounds of the set such that the aggregate gross mass of the reporter and linker combination is the same for each reagent compound of the set;
iv) X is a bond between an atom of the reporter moiety and an atom of the linker moiety;
v) Y is a bond between an atom of the linker moiety and an atom of the reactive group, wherein, once the reagent compound is reacted with the reactive analyte, bond Y links the linker to the analyte; and
vi) bonds X and Y are configured to fragment to form a labeled analyte when subjected to dissociative energy levels;
wherein the reporter moiety, RP, of each of the reagent compounds is independently a substituted or unsubstituted morpholine, or a substituted or unsubstituted piperazine, the nitrogen atom of the morpholine or a nitrogen atom of the piperazine is N-alkylated with a substituted or unsubstituted acetic acid moiety, the acetic acid moiety represents X-LK-Y, and the carbonyl group of the acetic acid moiety is linked to the reactive group, RG, to which the analyte can be linked, by reaction of the reactive group, RG, with the functional group of the analyte, and each of the two or more reagent compounds independently comprises one or more heavy atom isotopes.
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Abstract
This invention pertains to methods, kits and/or compositions for the determination of analytes by mass analysis using unique labeling reagents or sets of unique labeling reagents. The labeling reagents can be isomeric or isobaric and can be used to produce mixtures suitable for multiplex analysis of the labeled analytes.
35 Citations
15 Claims
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1. A set of two or more reagent compounds, or a salt thereof, each of the formula RP-X-LK-Y-RG, wherein:
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i) RG is a reactive group that is a nucleophile or an electrophile and that is capable of reacting with a functional group of one or more reactive analytes of a sample; ii) RP is a reporter moiety that comprises a fixed charge or that is ionizable, wherein the gross mass of each reporter moiety is different for each reagent compound of the set; iii) LK is a linker moiety that links the reactive group to the reporter moiety, wherein the mass of the linker compensates for the difference in gross mass between the reporter moieties for the different reagent compounds of the set such that the aggregate gross mass of the reporter and linker combination is the same for each reagent compound of the set; iv) X is a bond between an atom of the reporter moiety and an atom of the linker moiety; v) Y is a bond between an atom of the linker moiety and an atom of the reactive group, wherein, once the reagent compound is reacted with the reactive analyte, bond Y links the linker to the analyte; and vi) bonds X and Y are configured to fragment to form a labeled analyte when subjected to dissociative energy levels; wherein the reporter moiety, RP, of each of the reagent compounds is independently a substituted or unsubstituted morpholine, or a substituted or unsubstituted piperazine, the nitrogen atom of the morpholine or a nitrogen atom of the piperazine is N-alkylated with a substituted or unsubstituted acetic acid moiety, the acetic acid moiety represents X-LK-Y, and the carbonyl group of the acetic acid moiety is linked to the reactive group, RG, to which the analyte can be linked, by reaction of the reactive group, RG, with the functional group of the analyte, and each of the two or more reagent compounds independently comprises one or more heavy atom isotopes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13)
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14. A reagent compound of formula RP-X-LK-Y-RG, or a salt thereof, wherein:
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i) RG is a reactive group that is a nucleophile or an electrophile and that is capable of reacting with a functional group of one or more reactive analytes of a sample; ii) RP is a reporter moiety that comprises a fixed charge or that is ionizable; iii) LK is a linker moiety that links the reactive group to the reporter moiety; iv) X is a bond between an atom of the reporter moiety and an atom of the linker moiety; v) Y is a bond between an atom of the linker moiety and an atom of the reactive group, wherein, once the reagent compound is reacted with the reactive analyte, bond Y links the linker to the analyte; and vi) bonds X and Y are configured to fragment to form a labeled analyte upon reacting the compound of formula RP-X-LK-Y-RG with a sample comprising one or more reactive analytes to form a product and subjecting the product to dissociative energy levels;
wherein, the linker moiety of the compound is isotopically enriched with at least one heavy atom isotope and the reporter moiety of the compound is isotopically enriched with at least one heavy atom isotope;wherein the reporter moiety, RP, is a substituted or unsubstituted morpholine, or substituted or unsubstituted piperazine, the nitrogen atom of the morpholine or a nitrogen atom of the piperazine is N-alkylated with a substituted or unsubstituted acetic acid moiety, the acetic acid moiety represents X-LK-Y, and the carbonyl group of the acetic acid moiety is linked to the reactive group, RG, to which the analyte can be linked, by reaction of the reactive group, RG, with the functional group of the analyte. - View Dependent Claims (15)
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Specification