Releasable nonvolatile mass-label molecules
First Claim
Patent Images
1. A release tag compound comprising Rx, Re, and M wherein:
- Rx is a reactive group that specifically reacts with a target molecule;
Re is a release group for releasing M from Rx, and M is a nonvolatile mass label that identifies and characterizes the reactive group, wherein the mass label comprises a biopolymer detectable by mass spectrometry.
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Abstract
Releasable tag reagents for use in the detection and analysis of target molecules, particular in mass spectrometric analyses are provided. Also provided are methods of detection that employ releasable tag reagents.
231 Citations
44 Claims
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1. A release tag compound comprising Rx, Re, and M wherein:
- Rx is a reactive group that specifically reacts with a target molecule;
Re is a release group for releasing M from Rx, and M is a nonvolatile mass label that identifies and characterizes the reactive group, wherein the mass label comprises a biopolymer detectable by mass spectrometry. - View Dependent Claims (2, 3, 4, 5, 6, 7, 10, 11, 12, 13, 14, 15, 17, 18, 19, 34)
- Rx is a reactive group that specifically reacts with a target molecule;
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8. A release tag compound comprising Rx, Re, and M wherein:
- Rx is a reactive group that specifically reacts with a target molecule;
Re is a release group for releasing M from Rx; and
M is a nonvolatile mass label that identifies and characterizes the reactive group, whereina) the mass label comprises a synthetic polymer or a biopolymer detectable by mass spectrometry; b) the release group comprises a chemically cleavable linkage; and c) the chemically cleavable group is incorporated into the phosphate backbone. - View Dependent Claims (9)
- Rx is a reactive group that specifically reacts with a target molecule;
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16. The compound of 15, wherein the polypeptide comprises a product of a phage display method or a bacterial display method.
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20. A release tag compound comprising Rx, Re, and M wherein:
- Rx is a reactive group that specifically reacts with a target molecule;
Re is a release group for releasing M from Rx;
wherein Rx and Re are the same, and M is a nonvolatile mass label that identifies and characterizes the reactive group, wherein the mass label comprises a synthetic polymer or a biopolymer detectable by mass spectrometry.
- Rx is a reactive group that specifically reacts with a target molecule;
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21. A release tag compound comprising Rx, Re, and M wherein:
- Rx is a reactive group that specifically reacts with a target molecule;
Re is a release group for releasing M from Rx;
wherein Re is contained within Rx, and M is a nonvolatile mass label that identifies and characterizes the reactive group, wherein the mass label comprises a synthetic polymer or a biopolymer detectable by mass spectrometry.
- Rx is a reactive group that specifically reacts with a target molecule;
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22. A release tag compound comprising Rx, Re, and M wherein:
- Rx is a reactive group that specifically reacts with a target molecule;
Re is a release group for releasing M from Rx;
wherein Re is cleavable by an enzyme, and M is a nonvolatile mass label that identifies and characterizes the reactive group, wherein the mass label comprises a synthetic polymer or a biopolymer detectable by mass spectrometry. - View Dependent Claims (23, 24, 25, 26, 27, 28, 29, 30, 31, 32)
- Rx is a reactive group that specifically reacts with a target molecule;
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33. A release tag compound comprising Rx, Re, and M wherein:
- Rx is a reactive group that specifically reacts with a target molecule;
Re is a release group for releasing M from Rx; and
M is a nonvolatile mass label that identifies and characterizes the reactive group, wherein more than one mass label is incorporated and each mass label comprises a synthetic polymer or a biopolymer detectable by mass spectrometry.
- Rx is a reactive group that specifically reacts with a target molecule;
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35. A release tag compound comprising Rx, Re and M, wherein:
- Rx is a double-stranded oligonucleotide comprising a restriction endonuclease recognition site and Rx further comprises a self-complementary oligonucleotide hairpin;
Re is a release group comprising a phosphodiester linkage capable of being cleaved by a restriction endonuclease; and
M is a mass label detectable by mass spectrometry.
- Rx is a double-stranded oligonucleotide comprising a restriction endonuclease recognition site and Rx further comprises a self-complementary oligonucleotide hairpin;
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36. A release tag compound comprising Rx, Re, and M, wherein:
- Rx is a double-stranded oligonucleotide that specifically reacts with a target molecule and wherein hybridization of a portion of Rx to a target nucleic acid renders a portion of Rx single-stranded at Re;
Re is a chemically cleavable release group for releasing M from Rx; and
M is a mass label detectable by mass spectrometry that identifies and characterizes the double-stranded oligonucleotide;
wherein Re is located within Rx; and
wherein cleavage at the chemically cleavable release group is inhibited by the presence of a double-stranded oligonucleotide at said release group.
- Rx is a double-stranded oligonucleotide that specifically reacts with a target molecule and wherein hybridization of a portion of Rx to a target nucleic acid renders a portion of Rx single-stranded at Re;
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37. A set of release tag compounds, said set comprising one or more release tag compounds for detecting a particular target nucleic acid, each release tag compound comprising Rx, Re, and M, wherein:
- Rx is an oligonucleotide comprising a variable region and an invariant region that react with a target nucleic acid, wherein the oligonucleotide specifically reacts with a target nucleic acid, Rx further comprises a nucleotide or oligonucleotide attached thereto after hybridization to the target nucleic acid, and the added nucleotide or oligonucleotide comprises Re′ and
M′
, wherein;
Re′
is a release group; and
M′
is a mass label detectable by mass spectrometry;
Re is a release group for releasing M from Rx; and
M is a mass label detectable by mass spectrometry that identifies and characterizes the oligonucleotide.
- Rx is an oligonucleotide comprising a variable region and an invariant region that react with a target nucleic acid, wherein the oligonucleotide specifically reacts with a target nucleic acid, Rx further comprises a nucleotide or oligonucleotide attached thereto after hybridization to the target nucleic acid, and the added nucleotide or oligonucleotide comprises Re′ and
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38. A set of release tag compounds, said set comprising one or more release tag compounds for detecting a particular target nucleic acid, each release tag compound comprising Rx, Re, and M, wherein:
- Rx is an oligonucleotide comprising a variable region and an invariant region that react with a target nucleic acid, wherein the oligonucleotide specifically reacts with a tatget nucleic acid, Rx further comprises a nucleotide or oligonucleotide attached thereto after hybridization to the target nucleic acid, wherein the added nucleotide or oligonucleotide comprises a chain terminating moiety;
Re is a release group for releasing M from Rx; and
M is a mass label detectable by mass spectrometry that identifies and characterizes the oligonucleotide.
- Rx is an oligonucleotide comprising a variable region and an invariant region that react with a target nucleic acid, wherein the oligonucleotide specifically reacts with a tatget nucleic acid, Rx further comprises a nucleotide or oligonucleotide attached thereto after hybridization to the target nucleic acid, wherein the added nucleotide or oligonucleotide comprises a chain terminating moiety;
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39. A method of preparing a release tag compound, the method comprising:
- (a) obtaining a target nucleic acid sequence;
(b) synthesizing a reactive group capable of hybridizing to the target nucleic acid sequence;
(c) hybridizing the reactive group to the target nucleic acid sequence; and
(d) extending the reactive group to produce a release tag compound complementary to the target nucleic acid sequence, wherein the release tag compound comprises, in addition to the reactive group, a mass label detectable by mass spectrometry that identifies and characterizes the reactive group and a release group for releasing the mass label from the reactive group wherein the mass label comprises a biopolymer detectable by mass spectrometry. - View Dependent Claims (40)
- (a) obtaining a target nucleic acid sequence;
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41. A release tag compound comprising Rx, Re, and M wherein:
- Rx is a reactive group that specifically reacts with a target molecule and is a polypeptide;
Re is a release group for releasing M from Rx; and
M is a nonvolatile mass label that identifies and characterizes the reactive group, wherein the mass label comprises biopolymer detectable by mass spectrometry. - View Dependent Claims (42, 43, 44)
- Rx is a reactive group that specifically reacts with a target molecule and is a polypeptide;
Specification