Abstract: .alpha.-substituted derivatives of valproic acid are provided for conjugation to antigenic compositions, particularly poly(amino acids), and enzymes. The antigenic conjugates are employed for the production of antibodies, which find particular use in immunoassays for the determination of valproate, while the enzyme conjugate finds use in a homogeneous enzyme immunoassay for the determination of valproate. The compounds are synthesized by alkylating valproate at the tertiary carbon atom by an aliphatic chain with a terminal double bond which is cleaved to provide an acid or aldehyde group.

Abstract: Methods and compositions for assaying for coenzymes having N-substituted 1,4-dihydropyridyl as the active portion of the prosthetic group. The conversion of NAD to NADH is widely measured both for determining NAD-NADH (including the analogs thereof) dependent enzymes and enzymes that can be coupled to NAD-NADH dependent enzymes, and for determining ligands or receptors in competitive protein binding assays. Antibodies specific for NADH in the presence of NAD are employed, providing enhanced fluorescence over the normal fluorescence of unbound NADH. By determining the rate of formation of NADH or the concentration of NADH, the analyte or enzyme of interest can be determined.

Abstract: An assay method and compositions are provided for determining the presence of an analyte in a sample. The analyte is a member of an immunological pair (mip) of immunogens--ligand and receptor. The method has two basic elements: a solid surface to which one of the members of the immunological pair is bonded and a signal producing system, which includes a catalytic member bonded to a mip, which signal producing system results in a measurable signal on said solid surface related to the amount of analyte in the medium. The signal generating compound is produced without separation of the catalyst labeled mip bound to the solid surface from the catalyst labeled mip free in solution.

Abstract: Novel biological assay method for determining the presence of a specific organic material by employing a modified enzyme for amplification. By employing receptors specific for one or a group of materials (hereinafter referred to as "ligands") and binding an enzyme to the ligand or ligand counterfeit to provide an "enzyme-bound-ligand", an extremely sensitive method is provided for assaying for ligands. The receptor when bound to the enzyme-bound-ligand substantially inhibits enzymatic activity, providing for different catalytic efficiencies of enzyme-bound-ligand and enzyme-bound-ligand combined with receptor.The receptor, ligand and enzyme-bound-ligand are combined in an arbitrary order and the effect of the presence of ligand on enzymatic activity determined. Various protocols may be used for assaying for enzymatic activity and relating the result to the amount of ligand present.

Abstract: Method and compositions are provided for performing protein binding assays involving a homologous pair consisting of ligand and receptor for the ligand. The method employs a label conjugated to a member of said homologous pair and a uniformly dispersed discontinuous phase of discrete particles in a continuous aqueous phase, where the discrete particles create microenvironments which allow for discrimination between the label associated with the particle--in a discontinuous phase--and the label in the continuous phase.Various conjugates and particles are provided which find use in the subject method.

Abstract: Method and apparatus are provided for performing immunoassays employing a device comprising a relatively small test zone referred to as an immunosorbing zone, and a relatively large liquid absorbing zone in liquid receiving relationship with said immunosorbing zone. The immunosorbing zone includes a member of an immunological pair ("mip")--ligand and antiligand--bound to a support.A signal producing system is employed in conjunction with said device having as one component a signal label bound to a mip. The signal producing system provides for production of a detectible signal in the immunosorbing zone in relation to the amount of analyte in a sample.

Abstract: Fluorescent antigen conjugates are provided comprising antigens covalently bonded to at least one 2,7-dialiphatic substituted-9-phenyl-6-hydroxy-3H-xanthen-3-one, wherein the 1- and 8-positions are unsubstituted. Also provided are novel fluorescent compounds absorbing at wavelengths in excess of 500 nm, having active functionalities for linking to the antigen. Finally, methods are provided for analyzing antigens in serum, whereby serum interference is avoided.

Abstract: An improved method is provided for performing an enzyme immunoassay, where the signal producing compound is chemically labile. Particularly, in homogeneous enzyme immunoassays, where the enzyme is conjugated with a hapten, and an antibody binding to hapten reduces or inhibits the activity of the enzyme-hapten conjugate, the immunoassay is performed in accordance with conventional procedures. After a sufficient time, antienzyme is added which rapidly deactivates the enzyme, so that the signal remains constant for long periods of time.

Abstract: Novel polyhalothyronines or immunogenic mimetic analogs are provided conjugated to a fluorescer compound with the resulting conjugate being highly fluorescent. Specifically, the compounds have at least one of the following characteristics: bromine(s) at the 3',5'-positions of the thyronine or mimetic analog; or a rigid linking group between the thyronine and the fluorescer, so as to inhibit heavy atom catalyzed decay of fluorescence. The subject conjugates find a wide variety of uses in detecting for thyroid hormones as well as proteins which specifically bind to thyroid hormones.

Abstract: Compounds are provided which can be used in the preparation of reagents useful for quantitatively determining the presence of b-adrenergic blocking agents. These compounds contain a derivative of the blocking agent which is linked to an antigen or enzyme, the antigen conjugates being useful for preparing antibodies, and the enzyme conjugates being useful as detectors in an immunoassay as well. Additionally carboxylic acids, esters and aldehydes which are derivatives of the blocking agents are provided which are useful in preparing the above conjugates.

Abstract: Methods and compositions are provided for conjugating a wide variety of compounds, particularly polyfunctional compounds, having a mercapto group, either naturally present or synthetically introduced, to a polyamino compound, particularly a polypeptide (including proteins). The method employs a haloalkyl-carbonyl compound, which is conjugated to one or more of the amino groups under mild acylating conditions. This is followed by combining the acylated polyamino compound with a mercapto containing compound, whereby the halogen is displaced by the sulfur of the mercapto group to form a stable thioether linkage. The resulting conjugates, depending on the compounds involved, can find uses in immunoassays, as hapten-antigen conjugates for the production of antibodies, and as ligand analog enzyme conjugates for use as reagents in controlling the distribution of substitution of a mercapto compound to a polyamino compound.

Abstract: .alpha.-substituted derivatives of valproic acid are provided for conjugation to antigenic compositions, particularly poly(amino acids), and enzymes. The antigenic conjugates are employed for the production of antibodies, which find particular use in immunoassays for the determination of valproate, while the enzyme conjugate finds use in a homogeneous enzyme immunoassay for the determination of valproate. The compounds are synthesized by alkylating valproate at the tertiary carbon atom by an aliphatic chain with a terminal double bond which is cleaved to provide an acid or aldehyde group.

Abstract: Homogeneous immunoassay employing a light absorbing particle to which a member of an immunological pair (mip)-ligand and antiligand is bound and a labeled mip, where the label is light emitting. The labeled mip becomes bound to the light absorbing particle in proportion to the amount of analyte in the assay medium. Thus, the light emitted from the medium is related to the amount of analyte in the medium.

Abstract: Diether symmetrically substituted fluoresceins are provided having at least one anionic group and a linking functionality. Depending upon the site of substitution, the compounds can be used as fluorescers absorbing at wavelengths in excess of 500 nm or as quenchers, absorbing at wavelengths in excess of 500 nm and exhibiting substantially no fluorescence. The compounds find wide application, particularly as labels in fluorescent immunoassays.

Abstract: Amitriptyline functionalized compounds are provided for conjugation to antigenic compositions, particularly poly(amino acids), and enzymes. The antigenic conjugates are employed for the production of antibodies, which find particular use in immunoassays for the determination of amitriptyline, while the enzyme conjugate finds use in a homogeneous enzyme immunoassay for the determination of amitriptyline.

Abstract: An assay method and compositions are provided for determining the presence of an analyte in a sample. The analyte is a member of an immunological pair (mip) of immunogens--ligand and receptor. The method has two basic elements: a solid surface to which one of the members of the immunological pair is bonded and a signal producing system, which includes a catalytic member bonded to a mip, which signal producing system results in a measurable signal on said solid surface related to the amount of analyte in the medium. The signal generating compound is produced without separation of the catalyst labeled mip bound to the solid surface from the catalyst labeled mip free in solution.

Abstract: Novel carboxyalkylated disopyramides (NORPACE.RTM.) are provided as precursors for conjugating proteins, either antigenic for the preparation of antibodies or enzymatic for the preparation of enzyme conjugates, which antibodies and enzyme conjugates find use as reagents in immunoassays. The combination of antibodies and enzyme conjugates provide for sensitive, accurate, rapid assays for disopyramide without interference from closely analogous compounds, such as methadone or disopyramides metabolites.

Abstract: A method for determining a member of a specific binding pair-ligand and receptor (antiligand). Reagents employed include a first modified member which provides an electrical field due to the presence of a plurality of ionic charges and a second modified member labeled with a component of a signal producing system, which system may have one or more components. The average proximity in the assay medium of the first and second modified members is related to the amount of analyte, where the observed signal from the signal producing system is related to the effect of the electrical field on the signal producing system.Also, compositions are provided, as well as reagents, in predetermined ratios for optimizing the signal response to variations in analyte concentration.

Abstract: Novel biological assay method for determining the presence of a specific organic material by employing a modified enzyme for amplification. By employing receptors specific for one or a group of materials (hereinafter referred to as "ligands") and binding an enzyme to the ligand or ligand counterfeit to provide an "enzyme-bound-ligand", an extremely sensitive method is provided for assaying for ligands. The receptor when bound to the enzyme-bound-ligand substantially inhibits enzymatic activity, providing for different catalytic efficiencies of enzyme-bound-ligand and enzyme-bound-ligand combined with receptor.The receptor, ligand and enzyme-bound-ligand are combined in an arbitrary order and the effect of the presence of ligand on enzymatic activity determined. Various protocols may be used for assaying for enzymatic activity and relating the result to the amount of ligand present.

Abstract: Phencylidine, PCP, derivatives having a non-oxo carbonyl functionality linked directly or through a linking group to the phenyl ring are provided for conjugation to antigenic compositions, particularly poly(amino acids), and enzymes. The antigenic conjugates are employed for the production of antibodies, which find particular use in immunoassays for the determination of phencyclidine, while the enzyme conjugate finds use in a homogeneous enzyme immunoassay for the determination of phencyclidine.