Mass tags for quantitative analysis
OwnerRajendra Singh, Haihong Zhouinventors
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Abstract
The present invention relates generally to novel protein modification reagents for fractionation and quantitative (differential) profiling of proteins in a complex mixture. The reagents react with amino acids or other protein components or structures and function as mass tags. The present invention provides methods of making the protein modification reagents and methods of using the protein modification reagents for quantitative analysis of proteins.
Claims26
A protein mass tag (PMT) reagent for mass spectrometric analysis of proteins comprised of an amino acid reactive moiety that selectively reacts with certain protein functional groups, wherein said protein mass tag is differentially labeled with one or more non-isotopic chemical substituents.
A plurality of pmt reagents for mass spectrometric analysis of proteins each comprised of an amino acid reactive moiety that selectively reacts with certain protein functional groups, wherein each of said pmt reagents is differently labeled with one or more non-isotopic chemical substituents.
A plurality of PMT reagents for mass spectrometric analysis of proteins having the general formula:
- RM-PRM
- wherein RM is a recognition moiety and PRM is an amino acid reactive moiety that selectively reacts with certain protein functional groups, wherein each of said PMT reagents is differentially labeled with one or more non-isotopic chemical substituents.
17 A plurality of PMT reagents for mass spectrometric analysis of proteins having the general formula:
- RM-AM-PRM
- wherein RM is a recognition moiety, AM is an accessory moiety and PRM is an amino acid reactive moiety that selectively reacts with certain protein functional groups, wherein each of said PMT reagents is differentially labeled with one or more non-isotopic chemical substituents.
The PMT reagents of claim 17 wherein RM is biotin and AM is a fluorescent compound.
A compound having the following formula:
- 20
- wherein
- X is independently selected from the group consisting of H, D, OH, OD, R, OR, OSiR3, Cl, Br, I, F, SH, SR, NH2, NHR, and NR2;
- R is selected from the group consisting of an optionally substituted:
- C1-C20 alkyl, C2-C20 alkenyl, C2-C20 alkynyl, including deuterium substitutions;
- and
- n=0-10
A compound having the following formula:
- 21
- wherein
- X is independently selected from the group consisting of H, D, OH, OD, R, OR, OSiR3, Cl, Br, I, F, SH, SR, NH2, NHR, and NR2;
- R is selected from the group consisting of an optionally substituted:
- C1-C20 alkyl, C2-C20 alkenyl, C2-C20 alkynyl, including deuterium substitutions;
- and
- n=0-10.
A compound having the following formula:
- 22
A compound having the formula:
- 23
A method for identifying one or more proteins or protein components in one or more samples containing a mixture of proteins or protein components comprising:
- a) providing a plurality of PMT reagents wherein each PMT reagent is comprised of an amino acid reactive moiety that selectively reacts with certain protein functional groups, wherein each of said PMT reagents is differently labeled with one or more non-isotopic chemical substituents;
- b) contacting each sample with one of the PMT reagents to produce proteins or protein components in each sample labeled with a different PMT reagent;
- c) isolating said labeled proteins or protein components;
- and
- d) analyzing said labeled proteins or protein components.
A method for comparing two or more samples containing a mixture of one or more proteins or protein components comprising:
- a) providing a plurality of PMT reagents wherein each PMT reagent is comprised of an amino acid reactive moiety that selectively reacts with certain protein functional groups, wherein each of said PMT reagents is differently labeled with one or more non-isotopic chemical substituents;
- b) contacting each sample with a different PMT reagent to produce proteins or protein components in each sample labeled with a different PMT reagent;
- c) isolating said labeled proteins or protein components;
- and
- d) simultaneously analyzing said labeled proteins or protein components to quantitatively determine the relative amounts of protein or protein components in each sample.
Dependencies are inferred from the claim text.