Multisignal reagents for labeling analytes

Patent metadata

Field	Value
Publication number	US-10060910-B2
Application number	US-201213556304-A
Country	US
Kind code	B2
Filing date	Jul 24, 2012
Priority date	Jul 24, 2012
Publication date	Aug 28, 2018
Grant date	Aug 28, 2018

Abstract

Provided is a composition comprising an analyte bound to a polymer where the analyte is less than about 2000 MW and the polymer further comprises more than one signal or first member of a second binding pair. Also provided is an assay for an analyte comprising 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 competitive binding assay composition comprising:
(a) a sample suspected of containing an analyte;
(b) a detection reagent comprising
(i) the analyte or an analog thereof bound covalently to a polymer, or
(ii) the analyte or an analog thereof bound covalently to a first member of a first binding pair that binds to a polymer via a second member of the first binding pair which is covalently bound to the polymer,
wherein the analyte is not a member of the first binding pair, and
wherein the first binding pair is a sugar-lectin, an antigen-antibody, a ligand-ligand receptor, a hormone-hormone receptor, an enzyme-substrate, a biotin-avidin, or a biotin-streptavidin binding pair; and

(c) a binding agent capable of binding to the analyte or the analog thereof of the detection reagent and to analyte contained in the sample,
wherein the polymer further comprises more than one signal,
wherein said polymer is a nucleic acid and each signal comprises a fluorescent dye, a radioactive molecule, a chemiluminescent molecule or an enzyme,
wherein the second binding pair is a sugar-lectin, an antigen-antibody, a ligand-ligand receptor, a hormone-hormone receptor, an enzyme-substrate, a biotin-avidin, or a biotin-streptavidin binding pair, and
wherein the analyte is cAMP, cGMP, 8-bromoadenosine 3′,5′-cyclic monophosphate, cholesterol, a hydroxycholesterol, vitamin D, 25-hydroxy vitamin D, vitamin B12, vitamin E, vitamin B1, vitamin B6, ascorbic acid, retinol, biotin, folate, legumin, salbutamol, melamine, sulfaquinoxaline, an inositol phosphate, a phosphatidyl inositol phosphate, prednisone, pregnenolone, dexamethasone, triamcinolone, fludrocortisone, dihydrotachysterol, oxandrolone, testosterone, dihydrotestosterone, nandrolone, norethindrone, medroxyprogesterone acetate, progesterone, a glucocorticoid, aldosterone, estrogen, oxytocin, androstanediol glucuronide, bilirubin, warfarin, an estrogen, methotrexate, tobramycin, acetaminophen, encainide, fluoxetine, gentamycin, an aminoglycoside, amikacin, coenzyme Q10, theophylline, phenytoin, cimetidine, disulfiram, trazodone, ethanol, halothane, phenylbutazone, azapropazone, ibuprofen, amiodarone, imipramine, miconazole, metronidazole, nifedipine, chloramphenicol, trimethoprim, a sulfonamide, rifampin, cisplatin, vinblastine, bleomycin, oxacillin, nitrofurantoin, phenobarbital, primidone, carbamazepine, metoclopramide, cholestyramine, colestipol, neomycin, sulfasalazine, digoxin, indomethacin, diltiazem, erythromycin, tetracycline, itraconazole, nicardipine, triamterene, spironolactone, chlorpromazine, a cyclosporin, nortriptyline, ethosuximide, imipramine, codeine, lorazepam, topiramate, disopyramide, levetiracetam, clobazam, oxcarbazepine, fructosamine, caffeine, methaqualone, meprobamate, fluphenazine, a barbiturate, a phenothiazine, serotonin, valproic acid, digitoxin, maprotiline, lidocaine, mexiletine, primidone, risperidone, [OH-rispiridone] paliperidone, a porphyrin, haloperidol, flecainide, tocainide, acetazolamide, a sulfonamide, verapamil, a metanephrine, an oxidized nucleotide, a mycotoxin, tetrahydrocannabinol, a cannabinoid, cocaine, LSD, an amphetamine, heroin, methadone, nicotine, cotinine, a benzodiazepine, bis(2-ethylhexyl) phthalate, bisphenol A, hydralazine, atrazine, an organochloride insecticide, an organophosphate insecticide, or a carbamate insecticide.

2. The composition of claim 1 , wherein the detection reagent comprises:
(b)(ii) the analyte or an analog thereof bound covalently to a first member of a first binding pair that binds to a polymer via a second member of the first binding pair which is covalently bound to the polymer,
wherein the analyte is not a member of the first binding pair, and
wherein the first binding pair is a sugar-lectin, an antigen-antibody, a ligand-ligand receptor, a hormone-hormone receptor, an enzyme-substrate, a biotin-avidin, or a biotin-streptavidin binding pair.

3. The composition of claim 2 , wherein each of the first binding pair and the second binding pair is an antigen-antibody binding pair or a biotin-streptavidin binding pair.

4. A competitive binding assay comprising:
(a) a sample suspected of containing an analyte;
(b) a detection reagent comprising
(i) the analyte or an analog thereof bound covalently to a polymer, or
(ii) the analyte or an analog thereof bound covalently to a first member of a first binding pair that binds to a polymer via a second member of the first binding pair which is covalently bound to the polymer,
wherein the analyte is not a member of the first binding pair, and
wherein the first binding pair is a sugar-lectin, an antigen-antibody, a ligand-ligand receptor, a hormone-hormone receptor, an enzyme-substrate, a biotin-avidin, or a biotin-streptavidin binding pair;

(c) a binding agent capable of binding to the analyte or the analog thereof of the detection reagent and to analyte contained in the sample; and
(d) a linker moiety covalently linking
the analyte or analog thereof of the detection reagent to the first binding pair or the polymer, and/or
the polymer to at least one of the signals or first member of the second binding pair,

wherein the polymer further comprises more than one signal or more than one first member of a second binding pair of which the analyte is not a member,
wherein said polymer is a nucleic acid and each signal comprises a fluorescent dye, a radioactive molecule, a chemiluminescent molecule or an enzyme,
wherein the second binding pair is a sugar-lectin, an antigen-antibody, a ligand-ligand receptor, a hormone-hormone receptor, an enzyme-substrate, a biotin-avidin, or a biotin-streptavidin binding pair, and
wherein the analyte is cAMP, cGMP, 8-bromoadenosine 3′,5′-cyclic monophosphate, cholesterol, a hydroxycholesterol, vitamin D, 25-hydroxy vitamin D, vitamin B12, vitamin E, vitamin B1, vitamin B6, ascorbic acid, retinol, biotin, folate, legumin, salbutamol, melamine, sulfaquinoxaline, an inositol phosphate, a phosphatidyl inositol phosphate, prednisone, pregnenolone, dexamethasone, triamcinolone, fludrocortisone, dihydrotachysterol, oxandrolone, testosterone, dihydrotestosterone, nandrolone, norethindrone, medroxyprogesterone acetate, progesterone, a glucocorticoid, aldosterone, estrogen, oxytocin, androstanediol glucuronide, bilirubin, warfarin, an estrogen, methotrexate, tobramycin, acetaminophen, encainide, fluoxetine, gentamycin, an aminoglycoside, amikacin, coenzyme Q10, theophylline, phenytoin, cimetidine, disulfiram, trazodone, ethanol, halothane, phenylbutazone, azapropazone, ibuprofen, amiodarone, imipramine, miconazole, metronidazole, nifedipine, chloramphenicol, trimethoprim, a sulfonamide, rifampin, cisplatin, vinblastine, bleomycin, oxacillin, nitrofurantoin, phenobarbital, primidone, carbamazepine, metoclopramide, cholestyramine, colestipol, neomycin, sulfasalazine, digoxin, indomethacin, diltiazem, erythromycin, tetracycline, itraconazole, nicardipine, triamterene, spironolactone, chlorpromazine, a cyclosporin, nortriptyline, ethosuximide, imipramine, codeine, lorazepam, topiramate, disopyramide, levetiracetam, clobazam, oxcarbazepine, fructosamine, caffeine, methaqualone, meprobamate, fluphenazine, a barbiturate, a phenothiazine, serotonin, valproic acid, digitoxin, maprotiline, lidocaine, mexiletine, primidone, risperidone, [OH-rispiridone] paliperidone, a porphyrin, haloperidol, flecainide, tocainide, acetazolamide, a sulfonamide, verapamil, a metanephrine, an oxidized nucleotide, a mycotoxin, tetrahydrocannabinol, a cannabinoid, cocaine, LSD, an amphetamine, heroin, methadone, nicotine, cotinine, a benzodiazepine, bis(2-ethylhexyl) phthalate, bisphenol A, hydralazine, atrazine, an organochloride insecticide, an organophosphate insecticide, or a carbamate insecticide.

5. A competitive binding assay compos

Assignees

• Coleman Jack
• Szczepanik Maciej
• Jin Richard
• Enzo Life Sciences Inc

Inventors

• Coleman Jack
• Szczepanik Maciej
• Jin Richard

Classifications

• G01N33/54306 Primary

  Solid-phase reaction mechanisms · CPC title

• G01N33/58

  involving labelled substances (G01N33/53 takes precedence) · CPC title

• C12Q1/68

  involving nucleic acids · CPC title