Multisignal reagents for labeling analytes
Provided is a composition comprising an analyte bound covalently or through a first binding pair to a polymer. In this composition, the analyte is less than about 2000 MW; the polymer further comprises more than one signal or first member of a second binding pair; and the analyte is not a member of the first binding pair or the second binding pair. Also provided is an assay for an analyte. The assay comprises: combining a sample suspected of containing the analyte with the above-described composition and a binding agent that binds to the analyte; and detecting the signal or the first member of the second binding pair that is bound to the binding agent. Additionally provided is a multisignal labeling reagent comprising a first polymer covalently bound to (a) a reactive group or a first member of a first binding pair, and (b) more than one digoxigenin molecule.
1. A multisignal labeling reagent comprising:
a first polymer covalently bound to
(a) a reactive group or a first member of a first binding pair, and
(b) more than one digoxigenin molecule; and
bound to each digoxigenin molecule, an anti-digoxigenin antibody that is further bound to at least one signal molecule.
2. The multisignal labeling reagent of claim 1 , wherein each signal is a fluorescent dye, a radioactive molecule or an enzyme.
3. The multisignal labeling reagent of claim 1 , wherein each signal comprises an enzyme selected from the group consisting of alkaline phosphatase, horseradish peroxidase and luciferase.
4. The multisignal labeling reagent of claim 1 , wherein first polymer comprises a first member of a first binding pair, wherein the first binding pair is selected from the group consisting of a sugar-lectin, an antigen-antibody, a ligand-ligand receptor, a hormone-hormone receptor, an enzyme-substrate, a biotin-avidin, and a biotin-streptavidin.
5. The multisignal labeling reagent of claim 4 , wherein the first binding pair is biotin-streptavidin.
6. The multisignal labeling reagent of claim 1 , wherein the polymer comprises a nucleic acid, an abasic nucleic acid, a peptide nucleic acid, a polypeptide, a protein, an oligosaccharide, a polysaccharide or an organic polymer.
7. The multisignal labeling reagent of claim 1 , wherein the polymer is less than about 100 monomers.
8. The multisignal labeling reagent of claim 1 , wherein the polymer is a nucleic acid.
9. The multisignal labeling reagent of claim 8 , wherein the nucleic acid is DNA or comprises at least one nucleotide analog.
10. The multisignal labeling reagent of claim 8 , wherein the nucleic acid is less than about 100 nucleotides and/or nucleotide analogs.
11. The multisignal labeling reagent of claim 8 , wherein the nucleic acid is less than about 50 nucleotides and/or nucleotide analogs.
12. The multisignal labeling reagent of claim 8 , wherein the nucleic acid is less than about 25 nucleotides and/or nucleotide analogs.
13. The multisignal labeling reagent of claim 8 , wherein the nucleic acid is less than about 15 nucleotides and/or nucleotide analogs.
14. The multisignal labeling reagent of claim 8 , wherein the nucleic acid is linear.
15. The multisignal labeling reagent of claim 8 , wherein the nucleic acid is branched.
16. The multisignal labeling reagent of claim 1 , wherein the first polymer comprises a reactive group.
17. The multi signal labeling reagent of claim 16 , wherein the reactive group is isothiocyanate, isocyanate, monochlorotriazine, dichlorotriazine, mono- or di-halogen substituted pyridine, mono- or di-halogen substituted diazine, maleimide, aziridine, sulfonyl halogen substituted diazine, maleimide, sulfonyl halide, acid halide, hydroxysuccinimide ester, hydroxysulfosuccinimide ester, imido ester, hydrazine, azidonitrophenyl, azide, 3-(2-pyridyl dithio)-proprionamide, glyoxal, aldehyde, mercury salt, or any combination thereof.
18. The multi signal labeling reagent of claim 1 , further comprising a linker moiety covalently linking
the first polymer to the reactive group or first member of the first binding pair and/or
the first polymer to the more than one digoxigenin molecules.
19. The multi signal labeling reagent of claim 1 , wherein the first polymer is further bound to a second polymer, wherein the second polymer further comprises a signal molecule or a first member of a second binding pair.
20. The multisignal labeling reagent of claim 19 , wherein the first polymer is covalently bound to the second polymer.
21. The multisignal labeling reagent of claim 19 , wherein the first polymer is bound to the second polymer through a third binding pair.
22. The multi signal labeling reagent of claim 1 , bound to a target molecule through the reactive group or the first binding pair.
23. The multi signal labeling reagent of claim 22 , wherein the target molecule is a hapten, a protein, an oligopeptide, a nucleic acid, a nucleic acid analog, an oligosaccharide, a polysaccharide, a lipid or an organic polymer.