Abstract: The present invention relates to new compounds of the general formula: ##STR1## where X is iodine or hydrogen; A is a linking portion; and R is an iodinatable aryl or heteroaryl group having electron donating substituents. These compounds are useful precursors to the iodinated thyroid hormones for radioimmunoassay determination of thyroid hormones in biological fluids.

Abstract: A method of diagnosing Alzheimer's disease by the determination of the levels of the hormones somatotropin (human growth hormone) and somatomedin-C (IGF-I) after the administration of the L-dopa provocative test. Blood-sera samples are taken every thirty minutes for a two hour period after the administration of L-dopa, and the samples are tested for the levels of these hormones. These levels are then compared against the levels for normal subjects in the age group of between fifty and sixty. If the levels of the patient being diagnosed falls below the statistical mean corrected for error, then a diagnosis of Alzheimer's disease is indicated.

Abstract: A process of immunoassay of a selected substance in a liquid sample containing one or more other non-selected substances, each of these substances being comprised of at least two structurally different chains, the process comprising among others addition of a dissociating agent, such as for example a proteolytic enzyme, to the sample, mixing of the so-obtained liquid, inactivation of the dissociating agent, addition of antiserums, and determination of the amount of this selected substance in the liquid sample.

Abstract: Sacs comprising a compound having a hydrophilic peptide radical are prepared and are believed to have improved stability as a result of hydrogen bonding. The sacs, including a detectable marker and derivatized with a ligand may be used as a tracer in an assay.

Abstract: A derivative of 3,5,3'-triiodothyronine having the following general formula: ##STR1## in which m is an integer from 2 to 4, n is an integer from 1 to 4, and R is a monovalent, unsubstituted, fully unsaturated five- or six-membered monocyclic heterocyclic group containing either one or two nitrogen atoms as the only hetero atoms.

Abstract: A fluora immuno assay system. A fluorescent labeled assay reagent is prepared by conjugating an assay reagent with a fluorescent labeling agent. The fluorescent labeling agent is a chlorophyll or a porphyrin having a Stokes shift of not less than 150 nanometers. Apparatus for detecting the presence of the labeling agent comprising an excitation source illuminating a vessel with a photodetector directly within the illuminated area is also shown. The photodetector is insensitive to the spectrum of the excitation source.

Abstract: Compounds useful in a simultaneous multiple assay for analytes such as steroids, proteins, peptides, carbohydrates or drugs. The compound or compounds are prepared by labelling an individual analyte with a radioisotope through a chelating agent to form a coordinated compound. The assay uses one or more chelated labelled analytes with one or more labelled analytes wherein each radioisotope is different.

Abstract: Bifunctional aromatic compounds are employed as rigid coupling compounds for coupling one organic compound to another. For example, paranitrophenylisocyanate may be employed as a coupling compound for coupling thyroxine to a fluorescent dye to form a tracer for use in an assay.

Abstract: A method of performing an assay for an analyte comprises providing a chromatographic medium, preferably in sheet form, forming a reaction mixture containing a labelled reagent present partly in a form which is mobile on the medium and partly in a form which is immobile, the proportions of the two forms being related to the analyte concentration, applying the mixture to a spot on the chromatographic medium, optionally applying a solvent to cause the mobile form of the labelled reagent to migrate from the spot, and measuring the label intensity remaining at the spot. The method provides a convenient way of performing immunoassays, 2-site immunometric assays, and agglutination assays.

Abstract: An integral element for biological reaction comprising at least two layers consisting essentially of:a composite porous biological reaction layer comprising a fibrous material and a particulate material to which an active substance capable of participating in a biochemical reaction is fixed, anda porous layer of a fibrous material,in which:(1) the weight ratio between the particulate material and the fibrous material ranges from 1:20 to 1:0.3 on dry basis; and(2) the particulate material is contained in an amount ranging from 1 to 60 g/m.sup.2.

Abstract: Plastics surfaces are preactivated for immobilizing organo-chemical and biologic materials by coating the surfaces with a polypeptide which consists of a defined composition of hydrophobic amino acids and amino acids having side chains capable of activation and coupling. After activation of the plastics surfaces having been coated in this manner with various coupling reagents, it is possible to firmly fix organo-chemical and biologic materials. The materials having been immobilized in this manner are extremely stable to various reaction conditions.

Abstract: A specific binding enzyme-resistant ligand assay test material, which material comprises (a) a solid phase incorporated with one partner of a specific binding pair comprising said ligand or a binding analog thereof and a specific binding protein therefor; (b) a conjugate comprising the other partner of said specific binding pair incorporated with a substance which protects the specific binding protein of said pair from enzyme inactivation when bound with its partner; and (c) an active protein-inactivating enzyme. Also a specific binding method of assaying for an enzyme-resistant ligand in a sample, which method uses the above test material and which results in a reduction in interference caused by non-specific protein.

Abstract: The present invention relates to a simple and accurate method to be applied in immunoassay technique for separating the free ligand from the antibody-bound ligand.The method involves the use of a solvent extraction operation which at equilibrium provides the formation of two distinct phases the free- and bound-fraction. The solvent to be utilized should be slightly water miscible or completely water immiscible, having the property of marked selective extraction power toward the gamma-labelled constituent to be determined. The method is applicable in a liquid-liquid system, a solid-liquid system and solid-solid system.The method is useful for radioimmunoassay, free radical assay or metallo-immunoassay, being most versatile having also the advantage that as a result of the free ligand being extracted into the solvent, the determination of the quantity of gamma-labelled substance in the free and/or bound fraction compares very favorably with the known prior art methods in immunoassays.

Abstract: A method and apparatus for separation and filtration of particulate matter of sediment from a liquid sample. The system includes a unitary filter assembly having an intermediate fibrous filter material disposed between two layers of porous, relatively rigid plastic material. The unitary filter assembly is pressed axially toward the closed end of a sample container, such as a test tube, resulting in an upward flow of filtrate with compaction and retention of the particulates between the unitary filter assembly and the closed end of the sample container. The filter assembly is free of orientation criticality and may be readily adapted to automated handling in multiple tests or assays.

Abstract: Monoclonal antibodies specific for thyroxine (T.sub.4) are produced by two new and separate hybridoma cell lines. Combinations of the monoclonal antibodies from the two cell lines are used in an immunoassay for T.sub.4 of high accuracy over the range of T.sub.4 concentrations encountered in serum samples.

Abstract: A process for determining the presence of polyvalent antigens by incubation with three receptors is presented. In addition, a kit for carrying out this process is provided as well. One of said receptors is bound to a solid support and the other two, in solution, derive from the same animal species.

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: An improved radioimmunoassay test method and device based upon competitive binding test methods wherein immunoreactions are halted at a time when the rate of change of the quantity of bound radiolabeled analyte of interest is inversely proportional to the concentration of analyte of interest in an unknown sera. Based thereon, a test device is created having a single calibration curve 36 which is accurate throughout the shelf life of the device.

Abstract: Conjugates of heavy atoms containing analytes or their analogs and fluorescent molecules are covalently bonded to macromolecular supports to minimize the interference of fluorescence during assays, due to non-specific binding of serum proteins to the conjugate.

Abstract: A fast growing, continuous, functional rat thyroid cell strain, FRTL-5, which maintains functional characteristics of iodide uptake and thyroglobulin synthesis over prolonged periods of culture is cloned from FRTL cells obtained from primary cultures of Fischer rat thyroid glands. The FRTL-5 cells are cultured in a medium containing approximately 5 percent calf serum supplemented with a mixture of hormones, at least one of which is thyrotropin.The FRTL-5 cells are employed in a series of assays which measure thyroid stimulatory or inhibitory factors. The FRTL-5 system of assay specifically measures thymidine incorporation, cAMP elevation and iodide uptake and permits the evaluation of patient sera, particularly those afflicted with Graves' disease and other autoimmune thyroid diseases, thereby providing a means for determining appropriate methods of treatment.

Abstract: Isocyanate or isothiocyanate substituted lactone or thiolactone is employed for coupling one organic compound to another. For example, such coupling agents can be employed for coupling thyroxine or digoxin to a fluorescent compound for use as a tracer in an assay.

Abstract: Homogeneous assay for determining thyroid binding capacity by use of a fluorescent T.sub.3 tracer, such as T.sub.3 coupled to a fluorescein dye through a radical derived from L-lysine.

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: In the immunoassay of antigens in liquids, a reaction mixture is formed containing the liquid under assay, labelled antigen, and a mixed binding reagent which contains an antigen-binding site and a label-binding site, the two sites being spaced apart in the reagent so that a single molecule of labelled antigen cannot bind to both sites. The label is one whose activity is changed upon binding to a label-binding site, and the amount of antigen in the original liquid sample is determined by measuring the activity of the label in the reaction mixture. A preferred label is a fluorophore. The mixed binding reagent preferably consists of two antibodies linked together.

Abstract: Hybridoma tumor cell line (ATCC Number CRL 8164). A monoclonal anti-erythropoietin antibody substance, Epoclonalan, produced by said cell line. Use of Epoclonalan in immonological methods for isolation of natural erythropoietin and for quantitative detection of erythropoietin in fluid samples.

Abstract: In a particle agglutination assay for an antigen (Ag), there is included in the mixture a limited amount of a substance which binds univalently with a proportion of the Ag present, that Ag which is so bound being unable then to cause agglutination of the particles. In this way, unusually large concentrations of Ag can be assayed in that a proportion of the Ag is bound to the univalent substance and the particle agglutination assay is in effect conducted on the smaller amount of Ag still remaining free in solution.

Abstract: Competitive immunofluorescence assays for antigens in which immune complexes are precipitated with a nonfluorescent, nonlight-scattering precipitant such as polyethylene glycol and the resulting immunoprecipitate is dissolved with a nonfluorescent solvent of low ionic strength that maintains the pH of the solution substantially constant for immunofluorescence intensity reading. The assays are carried out by incubating the sample with fluorescent-labeled antigen, anti-antigen antibody, and a secondary antibody to the anti-antigen antibody followed by addition of the precipitant to form an immunoprecipitate. The precipitate is separated by centrifuging, dissolved in the solvent, and the immunofluorescence intensity of the solution is read with a fluorometer and compared to a standard curve.

Abstract: Ligand having at least two determinant or binding sites is contacted with a labeled first binder, an unlabeled second binder different than the first binder and a precipitating binder specific for the second binder, with such contacting precipitating a complex of the ligand and the first and second binders to thereby separate bound labeled binder from unbound labeled binder. The bound and/or unbound labeled binder is determined as a measure of the ligand.

Abstract: A process, means and devices for the quantitative determination, using electromagnetic radiation, of a component of a group including specifically bonding receptors and substances which can be specifically bonded by these receptors are based on a process comprising incubating one of the components, after it is immobilizing on the surface of a material having refractive index different from that of the components, with the other particular component and, optionally, further components, which can be labelled; irradiating the surface with electromagnetic radiation which is polarized parallel to the plane of incidence, the angle of incidence of the radiation being approximately equal to the angle .PHI. at which the intensity of the radiation reflected from the uncoated or precoated surface reaches a minimum; and ascertaining the intensity of the reflected radiation as a measure of the concentration to be determined.

Abstract: Methods are provided for reducing non-specific interference in competitive protein binding assays employing as the labeled reagent a fluorescent conjugate of a hydrophobic ligand conjugated to a fluorescer, which in turn is bound to a water soluble polysaccharide carrier ("fluorescer conjugate reagent"). In order to reduce non-specific interference from physiologic samples, the fluorescent reagent is combined with a lipid substituted neutral support, under conditions which provides two binding fractions: a first weakly binding fraction which is relatively free of non-specific interference in a competitive protein binding assay employing physiological fluids; and a second fraction, which more strongly binds to the lipid substituted support.

Abstract: Mixtures of monoclonal antibodies which contain effective assaying amounts of each of at least two monoclonal antibodies that bind to different antigenic sites on the antigen and are capable under appropriate conditions of binding simultaneously to an antigen are useful in enhanced sensitivity assays for the antigen. By utilizing such mixtures in diagnostic assays for important antigens such as the polypeptide human chorionic gonadotropin enhanced sensitivity can be achieved as compared with assays employing individual monoclonal antibodies.

Abstract: A method is disclosed for extraction, concentration, and bioassay measurement of the concentration of bioactive parathyroid peptides in biological or other fluids. Parathyroid peptides are extracted from the fluid by a support-antibody matrix specific to the N-terminal region. The bioassay is directed to the extracted parathyroid peptides. The volume of the medium containing the extracted parathyroid peptides and tissue, cells or tissue extracts with adenylate cyclase-coupled parathyroid hormone receptors may be chosen to be significantly less than the volume of biological or other fluid being assayed.

Abstract: Novel activated polyamide polymer container or test tube means and a method for the determination of ligand or analyte content in aqueous samples, are provided. The surfaces of the container tubes are specially activated thermally during their manufacture by injection molding thereby avoiding the need for subsequent coating of the tubes, e.g., with antibody, for binding assay procedures.

Abstract: A lectin is covalently bonded to an immunological conjugate such as an antibody-antigen or its equivalent. Then, the lectin-conjugate is isolated from the reaction product mixture by one of a number of alternative techniques involving one or more of the following types of reaction; (1) reversible reaction of the lectin with an insolubilized sugar to isolate lectin from the remainder of the mixture, (2) reaction of one immunological component (e.g., antibody) bonded to the lectin with an insolubilized corresponding component (e.g., antigen) to separate the antibody components from the remainder of the reaction mixture, and (3) filtration of the reaction components to separate on the basis of product molecular weight, size and/or shape of the components.

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: An immunoassay process for the detection of an antigen in a sample, which comprises: (a) forming a mixture of the sample with (1) a preformed complex of a primary antibody and a secondary binding macromolecule therefor, wherein the primary antibody is present at low concentrations and has substantial specificity for the antigen, the secondary binding macromolecule has substantial affinity for the Fc portion of the primary antibody, and the second binding macromolecule is affinity purified; and with (2) a detectably labeled form of the antigen; (b) incubating the mixture formed in step (a) for a time sufficient to allow the antigen and the detectably labeled antigen to competitively bind to the primary antibody of the preformed complex; (c) detecting the separated complex or the separated suspension medium.

Abstract: A method of forming an analyte-functionalized liposome for use in immunoassay techniques employs a phosphotriester intermediate of a phospholipid derivatized with the desired analyte in a process of forming a liposome which is functionalized on its outer surface with the analyte and which carries an enzyme marker. The method is also used to functionalize the liposome with a ligand which acts as a leash between the analyte and the liposome. Liposomes produced by the method include penicillin-G-functionalized liposome.

Abstract: Novel particle reagents for light scattering immunoassays are provided. The particle reagents are high refractive index shell-core polymers covalently bonded to compounds of biological interest or analogs thereof. A method of measuring unknown concentrations of these compounds of biological interest by measuring changes in turbidity caused by particle agglutination or its inhibition is also provided.

Abstract: Clinical method for evaulating thyrometabolic function by determination of clinically important thyroxine concentrations and thyroxine binding protein capacities in plasma.The method employs a competitive binding technique for determining free thyroxine concentration and thyroxine binding capacity for plasma having a known total thyroxine concentration.

Abstract: A method of determining the concentration of an antigen in a sample solution comprising(a) coating an antigen-protein conjugate onto a solid matrix,(b) conjugating an enzyme to an antibody specific for said antigen,(c) to a known quantity of a solution containing the antibody-enzyme conjugate of (b) adding a specified quantity of a sample containing an unknown amount of the antigen whose content is to be determined,(d) contacting the coated solid matrix of (a) with the solution (c) and incubate so as to effect binding between the antibody and antigen, some of the antigen being that from the sample and some being that on the solid matrix,(e) removing the solid matrix from the solution and washing,(f) immersing the solid matrix in a solution containing a known amount of an enzyme-substrate which is acted upon by the enzyme so as to effect reaction between the enzyme and enzyme-substrate to produce a product, and then separating the solid matrix from the solution of enzyme-substrate, and(g) then measuring the so

Abstract: Method and apparatus are provided for carrying out multiple simultaneous transfers of fluid. The method and apparatus are particularly directed toward immunoassays wherein immunologically active compounds, such as antigens and haptens, are detected through their associated antibodies. The device relies on the ability to transfer fluids, such as biological samples and reagents, between a reservoir and an associated receptacle. By providing a receptacle having a port at its lower end and which is otherwise hermetically sealed, such fluid transfer can be effected by immersing the port beneath the surface of the fluid in the reservoir and manipulating the pressure on the remaining surface area outside the port. The transfer of biological fluids at positive pressure provides enhanced fluids flow characteristics, particularly reduction or elimination of the tendency of these fluids to froth or bubble.

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: This invention relates to a highly-sensitive mini-iodinated radioactive hormone tracer which has both initial low iodine-to-hormone molar ratios and low specific activity. This invention also relates to methods for preparing and using such tracers in a radio-immunoassay which takes much less time than used heretofore and to a method of preparing said tracers.

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: An immunoassay for a specific antibody, particularly Graves' disease-specific antibody, in which interfering reactions by the reactive ends of similar antibodies are eliminated by the step of occluding the interfering reactive ends with an antibody against the interfering reactive ends.

Abstract: A method and kit for determining the presence of a polyvalent ligand in an aqueous fluid. The method involves incubating the fluid with an immobilized antibody to the ligand to form an immobilized ligand-antibody complex, then incubating the immobilized ligand-antibody complex with a solution of a soluble complex of a second antibody (to the ligand) and a labeled binding protein, such as protein A. The labeled binding protein is specifically bound to the Fc portion of the second antibody. The unbound second antibody and labeled binding protein are washed from the immobilized complex, and the presence of bound labeled binding protein is determined as a measure of the concentration of the ligand in the aqueous fluid.

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: A method for the determination of the thyroxine-binding index in serum, which method comprises mixing the serum sample to be determined with a defined amount of thyroxine and a defined amount of a determinable enzyme covalently bound to thyroxine, contacting the resulting solution with anti-thyroxine antibodies present in the solid phase, separating the liquid phase from the solid phase, and measuring the activity of said determinable enzyme in one of the phases as a measure of the tyroxine-binding index in said serum sample.

Abstract: This invention relates to a highly accurate, rapid and simple estimation of thyroxine (T.sub.4) directly from blood serum and also relates to the accurate measurement of triiodo-L-thyronine (T.sub.3) directly from blood serum. More specifically, the invention relates to a rapid, specific and reliable radioimmunoassay (RIA) technique for measurement of both T.sub.4 and T.sub.3 in unextracted serum. The method requires very small amounts of serum, e.g., 25 microliters (.mu.l) to measure T.sub.4 concentration in nearly all specimens representing clinical states of eu-, hypo- and hyperthyroidism, and 250 .mu.l to measure T.sub.3 concentrations in specimens representing most clinical states.