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. In this assay, the amount of the signal or the first member of the second binding pair bound to the binding agent is inversely proportional to the analyte in the sample. 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 composition comprising:
(i) a detection reagent comprising:
an analyte or an analog thereof bound covalently to a nucleic acid strand, the nucleic acid strand comprising
more than one signal, wherein each signal comprises a fluorescent dye, a radioactive molecule, a chemiluminescent molecule or an enzyme, and
one or more of abasic nucleic acids and peptide nucleic acids,
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-risperidone] 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; and
(ii) an antibody or antibody fragment that binds the analyte or analyte analog.
2. The composition of claim 1 , wherein the analyte is oxytocin, cAMP, cGMP, hydroxycholesterol, vitamin D, or 25-hydroxy vitamin D.
3. The composition of claim 2 , wherein the analyte is cAMP or vitamin D.
4. The composition of claim 1 , wherein the nucleic acid strand comprises abasic nucleic acids.
5. The composition of claim 2 , wherein the nucleic acid strand comprises abasic nucleic acids.
6. The composition of claim 3 , wherein the nucleic acid strand comprises abasic nucleic acids.
7. The composition of claim 1 , wherein the nucleic acid strand comprises peptide nucleic acids.
8. The composition of claim 2 , wherein the nucleic acid strand comprises peptide nucleic acids.
9. The composition of claim 3 , wherein the nucleic acid strand comprises peptide nucleic acids.
10. The composition of claim 1 , wherein each signal comprises a fluorescent dye or a chemiluminescent molecule.
11. The composition of claim 4 , wherein each signal comprises a fluorescent dye or a chemiluminescent molecule.
12. The composition of claim 7 , wherein each signal comprises a fluorescent dye or a chemiluminescent molecule.