Multisignal reagents for labeling analytes

Patent metadata

Field	Value
Publication number	US-10184934-B2
Application number	US-201715609360-A
Country	US
Kind code	B2
Filing date	May 31, 2017
Priority date	Apr 3, 2003
Publication date	Jan 22, 2019
Grant date	Jan 22, 2019

Abstract

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.

First claim

What is claimed is:

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.

Assignees

• Enzo Life Science Inc
• Enzo Life Sciences Inc

Inventors

• Coleman Jack
• Szczepanik Maciej
• Jin Richard

Classifications

• C09B11/24

  Phthaleins containing amino groups {; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes} · CPC title

• C07H21/04

  with deoxyribosyl as saccharide radical · CPC title

• G01N33/53 Primary

  Immunoassay; Biospecific binding assay; Materials therefor · CPC title

• C09B11/08

  Phthaleins; {Phenolphthaleins; Fluorescein} · CPC title

• G01N33/581

  with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates) · CPC title