Abstract: The present invention relates to a method which eliminates centrifugation and decantation steps, to be performed in an automatic manner for carrying out specific binding assay tests, wherein liquid and solid phases are present.According to the invention, use is made of a specially designed device, consisting of a mixing reservoir into which is fitted snugly a mixer separator having a channel in the vertical axis of the mixer-separator. A rack holding a number of said mixing reservoirs containing the incubated reagents and analytes, capped with the mixer separators, is placed into a press-device designed to perform at a controlled rate a downward movement. The mixer separators are pushed downwards into the mixing reservoirs at a chosen rate for a preselected distance to complete the mass transport and separation operations. The separation devices are removed and either one of the separated phases can be measured in the desired analytical instrument for a quantitative or qualitative determination.

Abstract: There is described an enzyme immunoassay for measuring the concentration of an analyte in a sample wherein the substrate for the enzyme forms at least a part of the sample. In a particular embodiment the sample comprises or consists of a milk sample and the enzyme is an enzyme capable of clotting milk. An example given of such an enzyme is chymosin. The assay described may be used to measure the concentration of progestagens or oestrogens in milk using the techniques of heterogeneous or homogeneous enzyme immunoassay. The results of such an assay give an indication of the fertility of a milk producing domestic animal (e.g. a cow) and may be used to diagnose pregnancy of such an animal. Particular compounds for use in the assay are described, as is a kit of reagents for use in the assay.

Abstract: A peptide having the formula ##STR1## is synthesized. The peptide is conjugated, e.g., with bis-diazotized benzidine, at its C-terminus to a carrier, such as bovine serum albumin, to form a synthetic antigen useful for inducing antibody production in a host animal. The antiserum obtained from the host animal is free of cross-reactivity with other pituitary substances and thus particularly advantageous for assay purposes. The peptide, whether unlabeled or labeled with radioactive iodine on the tyrosine moiety, has an affinity to antiserum raised against the conjugate similar to the affinity of natural hPRL to the antiserum. The synthetic peptide is used in radioimmunoassays where the labeled peptide competes for the binding sites in the antiserum with unknown concentrations of hPRL in biological samples.

Abstract: Methods and compositions are provided for detecting the presence of carcinomas in a mammalian host by measuring the level of normal surface antigens specific for a differentiated cell in the serum of the host as compared to the normal level of such antigen. The method finds particular use in detecting residual carcinomas after therapy or in detecting the recurrence of neoplastic tissue, and assigning a tissue of origin to the neoplastic tissue.

Abstract: Methods for the isolation and purification of an antigen, named NB/70K, from human ovarian carcinomas and radioimmunoassays for the detection of ovarian carcinomas, as well as an antibody specific for NB/70K.

Abstract: An immunoassay method for detection of antigen is disclosed. The method employs complement mediated lysis of vesicles loaded with In-111 or other gamma-emitting cation, and quantitative detection of the lysis by gamma-ray perturbed angular correlation (PAC) spectroscopy. The vesicles are labeled with a substance competitive to the antigen to be measured, and the concentration of the antigen in the sample measured by assessing the diminution in lysis due to the presence of the competing antigen.The method may also be used to assess the immunologic competence of a subject by injecting suitably sensitized vesicles and monitoring the in vivo lysis pattern by (PAC).

Abstract: An improved immunoassay for determining the presence of an analyte in a serum sample and conjugates therefor are described. The known method comprises combining the sample with a conjugate of the analyte and an enzyme and with a receptor for analyte, and determining the presence of analyte in the sample from the effect that the sample has on the enzymatic activity when compared to the enzymatic activity in the absence of analyte or in the presence of known amounts of analyte. The present invention resides in the conjugation to the enzyme label of a label protectant which allows modulation of the enzymatic activity to occur and minimizes background interference between the enzyme label and the sample, which would alter the enzymatic activity.

Abstract: A method of composition is featured for performing a fluoroimmunoassay of a biological fluid sample for an immunological reactant. A conjugate of a carrier labelled with fluorophores and coupled to an immunological reactant is mixed with the sample and a known quantity of binding agent. After equilibrium is accomplished, the bound and unbound portions are separated. The carrier in a selected portion is chemically treated to liberate the fluorophores. The fluorescent level is then measured and compared with a standard.

Abstract: A test kit and assay for detecting the presence of minute amounts of an organic reactant in a test sample includes a plurality of beads or other types of support structures that are impregnated with a fluorescer and coated with a ligand that specifically binds to the organic reactant being investigated. The beads are placed in an aqueous solution, together with the reactant which has been radiolabeled. The portion of the radiolabeled reactant that binds to the ligand is thereby brought in close enough proximity to the beads to activate the fluorescer to produce light energy. The radiolabeled reactant that does not bind to the ligand is, for the most part, disposed too far away from the beads to enable the radioactive energy emitted thereby to reach the fluorescer integrated into the beads. Thus, the level of light energy produced by the fluorescer is indicative of the amount of reactant present in the test sample.

Abstract: Method for fluorescence spectroscopic determination of a biological substance provided with a marker consisting of a lanthanide chelate complex formed by a lanthanide coupled to the substance via a chelate forming compound, such as an EDTA-analogue, the lanthanide ion before the detection is dissociated from the active substance for instance by adding a buffer with a pH value below 3.5 and a splitting detergent, whereafter the excitation in the determination takes place in the solution in the presence of a .beta.-diketone by using a short radiation pulse and the fluorescence from the marker is detected when the fluorescence from the noise sources has substantially ceased.

Abstract: Anti-Sm antibodies are produced by the hybridoma technique and employed to test for lupus erythematosus in humans.

Abstract: A hybridoma cell line is disclosed that secretes monoclonal antibodies which serve as a high titer, reproducible, biological reagent useful in biological/medical research for isolating and identifying phosphotyrosine-containing proteins. In addition, the antibodies have potential uses in diagnosis of a variety of diseases, including certain cancers. The antibodies, which have demonstrated affinity for a variety of molecules containing o-phosphotyrosine residues, were prepared using a synthetic analog, p-azobenzyl phosphonate (ABP) covalently linked to a carrier protein, as the antigen.

Abstract: Fluorescent conjugates are employed providing combinations of a fluorescent sensitizer and a fluorescent phycobiliprotein. The conjugates find use in applications where large Stokes shifts, high absorption coefficients and high fluorescence quantum yields are desired. Particularly, combinations of phycobiliproteins are employed where the wavelength of excitation may be 50 nm or more different from the emission wavelength.

Abstract: The use of anti-idiotype antibodies as functional substitutes for antigens or haptens in immunoassays is disclosed.

Abstract: The present invention describes an apparatus and method for conducting immunochemical reactions in a self-contained sealed unit that requires only the addition of an unknown sample and water. The apparatus comprises a test tube with at least three chambers each containing different chemicals, including a solid sphere, and separated from each other by a water-soluble barrier.

Abstract: A process for the determination of an antigen or hapten in homogeneous aqueous phase by incubation in the presence of antigen- or hapten-specific antibodies and of a definite amount of enzyme-marked antigen or hapten and measurement of the activity of the marker enzyme, wherein, after incubation, the reaction solution is heated under those temperature conditions and for a period of time at which the marker enzyme is inactivated by at least 50% in the absence of the antigen- or hapten-specific antibody and the enzyme activity thereafter measured.Also a reagent for carrying out this process, wherein it contains a definite amount of .beta.-galactosidase-hapten conjugate, hapten antibody, buffer (pH 6.0 to 8.5), a system for the determination of the .beta.-galactosidase activity and optionally a definite amount of .beta.-galactosidase antibody.

Abstract: A conjugate useful in determining the amount of antigen or antibody in a liquid sample, said conjugate having a marker, an immunoreactive component (i.e. antigen or antibody) bound to the marker and an insolubilizing binding component which is also bound to the marker. The insolubilizing binding component portion of the conjugate will react with an insolubilizing receptor to form a solid product of conjugate and receptor unless the conjugate reacts with the corresponding antigen or antibody to be analyzed in which event the conjugate will not react with the insolubilizing receptor. The conjugate will be added to a liquid sample containing an unknown amount of, for example, an antibody. A known amount of the corresponding antigen is also added which reacts with both the conjugate and antibody. After the reaction is complete, the liquid sample is contacted with the insolubilizing receptor.

Abstract: A method is provided to fluorometrically determine a ligand in an assay solution containing the ligand, reagent system and a fluorescer wherein the intensity of the fluorescer emitted by the assay solution is related to the change in the transmittive properties of the assay solution produced by the interaction of the ligand to be determined and a reagent system capable of producing a change in the transmittive properties of the assay solution in the presence of the ligand. In addition, novel reagent compositions are provided which may be utilized to either spectrophotometrically or fluorometrically determine the concentration of a ligand in an assay solution.

Abstract: Improvement in fluorescent polarization immunoassays for substances in blood plasma or serum comprising conducting the assays in dilute anionic surfactant solutions which disrupt the fluorescent bilirubin serum albumin complex without disturbing the antibody reaction in the immunoassay. In this manner, background fluorescence in icteric samples is greatly reduced.

Abstract: An improved heterogenous specific binding assay method 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 method is carried out using reagent means which comprises, as its labeled constituent, 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 system is utilized as means for monitoring the extent of binding of the labeled constituent in conventional heterogenous specific binding assay schemes. The presence of a ligand in a liquid medium may be determined following conventional competitive binding manipulative techniques.

Abstract: A chromogen, in particular, a fluorescent compound, is conjugated to a ligand through an amino acid spacer radical, which includes a free carboxyl group. The tracers are particularly suitable for use in a flow through assay for low molecular weight analytes. The tracers are rapidly bound, of good sensitivity, and low non-specific binding.

Abstract: Methods for determining the presence of antigens or antibodies in an aqueous sample or presence of antigens on the surface of cells. A preferred embodiment employs fluorescent antigens which compete with the sample antigens for antibody binding sites. The antibodies are deposited on a support surface means in alternating patterns. The surface means and fluorescence detector are translocated with respect to each other and a signal generated by the detection of the repeating pattern of fluorescence. The signal is analyzed by means of a gated integrator responsive to a gate track control means also located on the surface means. Immunoassay methods having increased sensitivity are thereby obtained.

Abstract: "Two-site" or "sandwich" immunometric assay techniques for determination of the presence and/or concentration of antigenic substances in fluids using monoclonal antibodies are described and compared to conventional assays using polyclonal antibodies. Also described are inhibition assays using complexes of antigens with a monoclonal antibody.

Abstract: An immunoassay method utilizes antigen tagged, enzyme encapsulating liposomes which are immunospecifically ruptured in the presence of cognate antibody and active complement. A homogeneous phase reaction occurs with the antibody and complement acting to release the enzyme if an immunospecific antigen-antibody complex is formed at the surface of the liposome. The positions of the antigen and antibody can be reversed.

Abstract: This disclosure relates to a method and reagents for determining ligands in biological fluids such as serum, plasma, spinal fluid, amnionic fluid and urine. This disclosure also relates to a novel class of tracer compounds employed as reagents in fluorescence polarization immunoassays.

Abstract: This invention encompasses a method for measuring unsaturated thyroxine binding protein sites in a sample comprising intermixing with the sample an effective amount of fluorescent labeled tracer having binding affinity for thyroxine binding proteins and then measuring the amount of tracer bound to the thyroxine binding protein using fluorescence polarization techniques.

Abstract: An analysis film comprises a reagent layer composed of a porous material which contains an antibody but does not substantially contain a complex of an analyte or a labelled antigen with the antibody. In the analysis film, reagents for enzyme immune reaction of homogenous type are incorporated so that an analyte is analyzed without requiring B/F separation. An analysis method for various analytes using the same provides high sensitivity, high accuracy as well as good reproducibility and is simple and rapid.

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: Reagent mixtures for single test which allow for rapid determination of drugs without sophisticated equipment. Into a single vial as dry powders are combined an enzyme bound ligand reagent, an antiligand reagent (antibody), appropriate substrates, bulking agents, as well as other additives. Upon addition to the reagent mixture of an appropriate volume of diluent and the sample suspected of containing the drug, optionally subject to prior treatment and/or dilution, the reagents are activated and either a single reading at a predetermined time interval or two or more readings over a predetermined time interval are taken of spectrophotometric changes in the solution. By comparison to a standard, the concentration of the drug may be determined quantitatively.

Abstract: A method for determining the presence of a ligand in, or the ligand binding capacity of a liquid test sample which includes the steps of (a) adding to the sample a conjugate of the ligand and a label, (b) contacting the sample with a test device containing reagents which in conjunction with the conjugate and ligand, are capable of producing a detectable response, and (c) measuring the response.

Abstract: An immunoassay for analytes such as antigens or haptens, which utilizes covalent hybrid antibodies to modulate the activity of indicators. The hybrid antibody has binding sites for the analyte and the indicator. Final activity of the indicator is proportional to analyte concentration.

Abstract: A technique for measuring the degree of an antigen-antibody reaction by preparing a suspension of insoluble microscopic carrier particles of at least one type carrying an antigen, an antibody or a hapten, forming an agglutination promoting or inhibiting reaction system among the insoluble carrier particles based on an antigen-antibody reaction using the suspension and one or more antigen, antibody or hapten, irradiating the solution of the reaction system with laser light and detecting the light scattered from the reaction system at one or more specific angles, detecting a signal indicative of one or more specific frequency bands from the resulting scatter spectrum, and thenceforth calculating the quantity of antigen, antibody or hapten in a specimen on the basis of the detected signal. The intensity spectrum output of filter 11 for frequency band selection is in the form of a square root and is converted into the original intensity spectrum by means of a squaring circuit 12.

Abstract: A method of measuring the degree of partitioning of a labeled species between free and bound states which involves the use of an insoluble porous monolith having a means for binding a portion of the labeled species within the pores thereof, which monolith is capable of substantially attenuating the signal emitted by labeled species subsequently bound within the pores thereof.

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 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 the sample.

Abstract: Species-linked diamine triacetic acids of the formula ##STR1## wherein T is an organic species containing at least one amine, hydroxyl, or thio functional group, L is the residue of at least one of those functional groups and R is a two or more atom long covalent bridge, are disclosed. Methods for their preparation, for the preparation of metal chelates from them and for the use of the chelates are also disclosed. In a preferred embodiment, the metal ions employed in the formation of the chelates are rare earth metal ions capable of forming fluorescent chelates which can in turn be employed in fluoroassay techniques.

Abstract: In the course of a reaction in which one of the reactants is on the surface of carrier particles in a solution and another of the reactants is tagged with a fluorescent substance, some of the fluorescently tagged reactant attaches to, or is displaced from the carrier particle. The present invention relates to a method and device for determining the amount of fluorescently-tagged reactant which is attached to the carrier particle or which is free in solution, without physically separating the carrier particles from the solution. In a particular application of the invention (immunoassay) the reaction is between antibodies and antigens, and from the amount of fluorescently-tagged reactant which is attached to the carrier particle one can determine the unknown amount of antigen in a sample.

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: In the course of a reaction in which one of the reactants is on the surface of carrier particles in a solution and another of the reactants is tagged with a fluorescent substance, some of the fluorescently tagged reactant attaches to, or is displaced from the carrier particle. The present invention relates to a method and device for determining the amount of fluorescently-tagged reactant which is attached to the carrier particle or which is free in solution, without physically separating the carrier particles from the solution. In a particular application of the invention (immunoassay) the reaction is between antibodies and antigens, and from the amount of fluorescently-tagged reactant which is attached to the carrier particle one can determine the unknown amount of antigen in a sample.

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: This invention relates to a method and to apparatus for chemical and biochemical analyses which employ an energy-transmissive core and may employ one or more sheaths which selectively absorb, react with, and/or filter an analyte or a product of an analyte. The core is transmissive to a chosen energy carrier and it has an inlet end and an outlet end. Between these ends it has an extended length. The passage of energy through the core is modified by reason of events which occur in one or more of the sheaths or in the case where no sheath is employed, by reason of events which occur in an ambient fluid. The resulting modification of the transmitted energy is a measure of such events which in turn are a measure of the analyte. The energy may be any of several types of energy which can be transmitted through the core from end to end and which is susceptible to modification by reactions in the sheath or sheaths or ambient fluid. The energy may be electromagnetic, electrical or sonic.

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: An indirect, non-destructive, quantitative assay for the presence of antigens or antibodies in a biological fluid. The assay is based on the interfunctional behavior of a known biological material with the material whose presence is quantitatively sought. In the assay, the known biological material by a correlative action links two discreet interfunctional particles together within a zone of activation with one of the particles emitting light-pulses upon bombardment of electrons from the other particle within the zone of activation. Depending upon which presence is sought to be measured, i.e.; antigen or antibody, the detectable light-pulses give one measurement which gives the ultimate quantitative measurement of the presence of either an antibody or antigen in the biological fluid after their initial interfunctional behavior one to the other. The interfunctional behavior itself is conventional and known in the art.

Abstract: A system for the detection of a biological analyte of interest is disclosed which comprises a microencapsulated fluorescer material which has been conjugated to an immunological specie specific to the biological analyte of interest, a means of disrupting the capsule containing the fluorescer and an energy source other than electro-magnetic radiation which is capable of activating the fluorescer.