MULTISIGNAL REAGENTS FOR LABELING ANALYTES

• US 20170315113A1
• Filed: 05/31/2017
• Published: 11/02/2017
• Est. Priority Date: 04/03/2003
• Status: Active Grant

• Current Assignee: Enzo Life Sciences Incorporated (Enzo Biochem Incorporated)
• Original Assignee: Enzo Life Science, Inc.
• Inventors: Coleman, Jack, Szczepanik, Maciej, Jin, Richard
• Granted Patent: US 10,184,934 B2
• Time in Patent Office: Days
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C09B 11/08 Phthaleins; Phenolphthaleins; Fluorescein
• CPC Class Codes: C09B 11/24 Phthaleins containing amino groups ; Phthalanes; Fluoranes; Phthalides; Rhodamine dyes; Phthaleins having heterocyclic aryl rings; Lactone or lactame forms of triarylmethane dyes
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/581 with enzyme label (including co-enzymes, co-factors, enzyme inhibitors or substrates)
• CPC Class Codes: G01N 33/82 involving vitamins or their receptors
• CPC Class Codes: G01N 33/94 involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors

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.

Claims

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.

2. The multisignal labeling reagent of claim 1, wherein each digoxigenin molecule is bound to an anti-digoxigenin antibody that is further bound to at least one signal molecule.

3. The multisignal labeling reagent of claim 2, wherein each signal is a fluorescent dye, a radioactive molecule or an enzyme.

4. The multisignal labeling reagent of claim 2, wherein each signal comprises an enzyme selected from the group consisting of alkaline phosphatase, horseradish peroxidase and luciferase.

5. 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.

6. The multisignal labeling reagent of claim 5, wherein the first binding pair is biotin-streptavidin.

7. 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.

8. The multisignal labeling reagent of claim 1, wherein the polymer is less than about 100 monomers.

9. The multisignal labeling reagent of claim 1, wherein the polymer is a nucleic acid.

10. The multisignal labeling reagent of claim 9, wherein the nucleic acid is DNA or comprises at least one nucleotide analog.

11. The multisignal labeling reagent of claim 9, wherein the nucleic acid is less than about 100 nucleotides and/or nucleotide analogs.

13. The multisignal labeling reagent of claim 9, wherein the nucleic acid is less than about 25 nucleotides and/or nucleotide analogs.

14. The multisignal labeling reagent of claim 9, wherein the nucleic acid is less than about 15 nucleotides and/or nucleotide analogs.

15. The multisignal labeling reagent of claim 9, wherein the nucleic acid is linear.

16. The multisignal labeling reagent of claim 9, wherein the nucleic acid is branched.

17. The multisignal labeling reagent of claim 1, wherein the first polymer comprises a reactive group.

18. The multisignal labeling reagent of claim 17, 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.

19. 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.

20. 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.

21. The multi signal labeling reagent of claim 1, wherein the first polymer is covalently bound to the second polymer.

22. The multi signal labeling reagent of claim 1, wherein the first polymer is bound to the second polymer through a third binding pair.

23. The multi signal labeling reagent of claim 1, bound to a target molecule through the reactive group or the first binding pair.

24. The multi signal labeling reagent of claim 23, 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.

12. The multisignal labeling reagent of claim 594, wherein the nucleic acid is less than about 50 nucleotides and/or nucleotide analogs.