Abstract: Procainamide and N-acetylprocainamide (NAPA) immunogens, antibodies prepared therefrom, labeled conjugates, synthetic intermediates, and the use of such antibodies and labeled conjugates in immunoassays for determining the respective drugs. The immunogens comprise the drugs coupled at the .alpha.-position of the amide side chain to an immunogenic carrier material. The labeled conjugates and synthetic intermediates similarly are .alpha.-position derivatives of the drugs or precursors thereof. The antibodies and labeled conjugates are particularly useful in homogeneous nonradioisotopic immunoassays for measuring the respective drugs in biological fluids such as serum.

Abstract: A homogeneous specific binding assay device, a method for its preparation, and a method for its use in determining a ligand, such as antigen, hapten, or antibody, in, or the ligand binding capacity of, a liquid sample. The test device comprises a solid carrier member, such as a fibrous web matrix, e.g., paper, or a polymeric film or gel, incorporated with reagents for a homogeneous specific binding assay system which produces a detectable response, usually an electromagnetic radiation signal, that is a function of the presence or amount of the ligand in or the ligand binding capacity of the sample. Useful homogeneous specific binding assay systems include those involving enzyme substrate labels, enzyme prosthetic group labels, and enzyme labels. The detectable response preferably is a luminescent, fluorescent, spectrophotometric, or colorimetric response, which is measurable by visual observation or instrument means.

Abstract: Immunogens for preparing antibodies against the drug lidocaine and related compounds, labeled conjugates, synthetic intermediates, and the use of such antibodies and labeled conjugates in immunoassays for determining lidocaine and such related compounds. The immunogens comprise lidocaine or an analog thereof coupled through one of the aromatic methyl groups to a conventional immunogenic carrier. The antibodies and labeled conjugates are particularly useful in homogeneous nonradioisotopic immunoassays for measuring lidocaine or its analogs in biological fluids such as serum.

Abstract: Disopyramide immunogens, antibodies prepared therefrom, labeled conjugates, synthetic intermediates, and the use of such antibodies and labeled conjugates in immunoassays for determining disopyramide. The immunogens comprise disopyramide coupled by an ether linkage at a selected singular position on its phenyl ring to an immunogenic carrier material. Likewise, the labeled conjugates and synthetic intermediates are phenyl ether derivatives of disopyramide or precursors thereof. The antibody and labeled conjugates are particularly useful in homogeneous nonradioisotopic immunoassays for measuring disopyramide in biological fluids such as serum.

Abstract: An immunoassay method and reagent system for determining nonenzymatically glucosylated proteins and protein fragments in a biological fluid based on the specific binding of such proteins and fragments with anti(Amadori-rearranged glucose), e.g., antibodies which selectively recognize the rearranged deoxyfructose form of glucose resulting when proteins are nonenzymatically glucosylated. The antibodies are raised against an immunogen comprising an immunogenic carrier material bearing 1-deoxy-1-fructosyl residues or conformers of such residues. Measurement of nonenzymatically glucosylated proteins and fragments thereof provides a useful index of blood glucose levels.

Abstract: A test composition, device, and method for their use in a homogeneous specific binding assay which employs a substance having reactant activity, i.e., a reactant, as a labeling substance in the detection of a ligand in a liquid medium. The test composition and device comprise a conjugate formed of a specific binding substance coupled to the reactant. The reactant advantageously is an enzymatic reactant such as an enzyme substrate or coenzyme. The activity of the conjugated reactant as a constituent of a predetermined reaction is affected by reaction between the specific binding substance in the conjugate and a specific binding counterpart thereto. The presence of a ligand in a liquid medium may be determined using competitive or displacement binding or sequential saturation techniques wherein the specific binding substance in the conjugate is the ligand or a specific binding analog thereof, or using a direct binding technique wherein the specific binding substance is a specific binding partner of the ligand.

Abstract: An improved immunoassay method, reagent means, test kit, and test device for determining an iodothyronine, e.g., thyroxine (T-4), in a biological fluid, usually serum or plasma, wherein 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid (HMS), or a salt thereof, is employed as a blocking agent for the binding of iodothyronines to thyroxine binding protein (TBP). The present invention is particularly advantageous as applied to homogeneous competitive binding iodothyronine immunoassays employing labels which are participants in enzyme-catalyzed reactions. Such labels include enzyme substrates, coenzymes, enzyme inhibitors, enzyme prosthetic groups, and enzymes.

Abstract: Immunoassay method and reagent means for determining theophylline in biological fluids such as serum. The antibody employed in the method is prepared from an immunogen comprising theophylline coupled at its 9-position to an immunogenic carrier material.

Abstract: Nucleic acid hybridization assay methods and reagent systems for detecting a particular polynucleotide sequence in a test medium. An aggregate is formed in the assay reaction mixture comprising intercalation complexes between a nucleic acid intercalator and double stranded nucleic acid associated with the hybridization product of the sequence to be detected and a nucleic acid probe sequence. Hybridization of the probe with the sequence to be detected can then be determined by addition of an antibody, or a fragment thereof, capable of binding with the intercalation complexes in the formed aggregate and measuring the antibody or fragment thereof which becomes bound to such intercalation complexes associated with hybridized probe. In one preferred embodiment, this method eliminates the need to chemically modify the probe in order to form a labeled reagent. In another embodiment, the method provides an advantageous method for labeling the probe by chemical modification.

Abstract: Fluorogenic indicator compounds responsive to the presence of hydrogen peroxide and methods and test compositions for determining hydrogen peroxide in a test sample. Preferred indicator compounds are 3,4-dihydrocoumarin derivatives and 3,4-dihydro-2-quinolone derivatives, especially those having a dialkylamino group at the 7-position. Such particularly preferred compounds are of the formula: ##STR1## wherein R.sup.1 is hydrogen, cyano, or --COOR.sup.6, --CONHR.sup.6 or --CON(R.sup.6).sub.2 where R.sup.6 is hydrogen, alkyl, alkenyl, or aryl; R.sup.3 and R.sup.4 are lower alkyl; and Y is .dbd.O or .dbd.N--R.sup.5 where R.sup.5 is hydrogen or lower alkyl. The indicator compounds yield highly fluorescent products upon oxidation by hydrogen peroxide in the presence of a peroxidatively active substance and are useful in analytical systems which generate hydrogen peroxide in response to an analyte under determination in a test sample.

Abstract: Theophylline immunogens, antibodies prepared therefrom, labeled theophylline conjugates, synthetic intermediates, and the use of such antibodies and labeled conjugates in immunoassays for determining theophylline. The immunogens comprise theophylline coupled at its 9-position to an immunogenic carrier material. Likewise, the labeled conjugates and synthetic intermediates are 9-position derivatives of theophylline or synthetic precursors thereof. The antibody and labeled reagents are particularly useful in homogeneous nonradioisotopic immunoassays for measuring theophylline in biological fluids such as serum.

Abstract: Tricyclic antidepressant drug (e.g., imipramine, desipramine, amitriptyline, or nortriptyline) immunogens, antibodies prepared therefrom, labeled conjugates, synthetic intermediates, and the use of such antibodies and labeled conjugates in immunoassays for determining such drug. The immunogens comprise the drug coupled at its 2'-position to an immunogenic carrier material. Similarly, the labeled conjugates and synthetic intermediates are 2'-derivatives of the drug or a precursor thereof. The antibodies and labeled reagents are particularly useful in certain homogeneous nonradioisotopic immunoassays for measuring tricyclic antidepressant drugs in biological fluids such as serum.

Abstract: A method for increasing the ability of apoglucose oxidase to combine with flavin adenine dinucleotide (FAD) and derivatives thereof to form active glucose oxidase by interacting apoglucose oxidase with an immunologically derived binding substance, e.g., an antibody or a fragment thereof, having a specific binding affinity for glucose oxidase. The apoglucose oxidase/anti-glucose oxidase immune complex is characterized by enhanced ability to combine with FAD and FAD-derivatives to yield glucose oxidase activity. The activation is particularly significant at temperatures elevated from room temperature, e.g., between 30.degree.-45.degree. C. An improved homogeneous specific binding assay method is provided for determining ligands wherein an FAD label is used and is monitored by its ability to combine with apoglucose oxidase to form active glucose oxidase by including anti-glucose oxidase in the reaction mixture.

Abstract: Immunogen conjugates comprising amino-functionalized haptens coupled through a carbonyl bridge to amino groups in immunogenic proteins and polypeptides. The resulting simple urea linkage in the conjugate is hydrophilic and contributes essentially no haptenic determinants. The immunogen is characterized by a high epitopic density. The method involves reaction of the hapten with a carbonyl diimidazole followed by addition of the protein or polypeptide carrier. Protein crosslinking problems common to prior art coupling methods are avoided.

Abstract: A propranolol immunogen comprising an unsubstituted hydroxypropylamine precursor of propranolol coupled through a bis-imidate linking group to an immunogenic carrier material. The immunogen is useful to stimulate the production of antibodies which recognize propranolol and can perform as reagents in immunoassays to determine propranolol.

Abstract: Immunogen conjugates comprising N-substituted-amino-3-desoxydigoxigenin derivatives coupled to conventional immunogenic carrier materials, and antibodies raised against such conjugates. Also provided are labeled digoxigenin conjugates for use with the digoxigenin antibodies in preferred immunoassay techniques for determining digoxin in biological fluids.

Abstract: An improved immunoassay method, reagent means, and test kit for determining an iodothyronine, e.g., thyroxine (T-4), in a biological fluid, usually serum or plasma, wherein fenclofenac and related phenylacetic acids, or salts thereof, are employed as novel blocking agents for the binding of iodothyronines to thyroxine binding protein (TBP). The present invention is particularly advantageous as applied to homogeneous competitive binding iodothyronine immunoassays wherein a spectrophotometric response is generated in the assay reaction mixture at a wavelength greater than about 300 nm, the blocking agents of the present invention having been found to have no substantial absorbance at wavelengths above 300 nm. Such homogeneous immunoassays include those which employ labels such as fluorescers, enzyme substrates, enzyme prosthetic groups, enzymes, and enzyme inhibitors.

Abstract: Intermediates in the preparation of 9-position derivatives of theophylline useful in synthesizing theophylline immunogens and labeled conjugates. The labeled conjugates and the antibodies produced against the immunogens are particularly useful in homogeneous nonradioisotopic immunoassay for measuring theophylline in biological fluids such as serum.

Abstract: An improved specific binding assay method and reagent for determining a ligand in a liquid medium employing, as an enzyme-cleavable substrate label, a residue having the formula:G--D--Rwherein G is a glycone, D is a dye indicator moiety, and R is a linking group through which the label residue is covalently bound to a binding component of a conventional binding assay system, such as the ligand, an analog thereof, or a specific binding partner thereof. The monitored characteristic of the label is the release of a detectable product, usually a fluorogen or chromogen, upon enzymatic cleavage of the glycosidic linkage between the glycone and the dye indicator moiety. The assay method may follow a homogeneous or heterogeneous format. The preferred glycone is a .beta.-galactosyl group and the preferred dye indicator moiety is an umbelliferone residue.

Abstract: Immunogen conjugates comprising N-aminoalkyl derivatives of iodothyronines, e.g., thyroxine and its lower alkyl esters, coupled to an immunogenic carrier material, and antibodies raised against such immunogen conjugates.