Abstract: Carcinoembryonic antigen contains immune determinates in common with .alpha.-acid glycoprotein. Carcinoembryonic antigen-containing compositions are purified by adsorption onto antibody to .alpha.-acid glycoprotein. The purified compositions may be employed as standards in carcinoembryonic antigen assays or in the labelled form as tracers. Intact carcinoembryonic antigen is assayed by a modified sandwich-type immunoassay. Other cancer-associated substances may be identified by searching for high molecular weight analogues of normal proteins.

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: A method of reacting a first reactive substance bonded to the inner wall surface of a reaction vessel and a second reactive substance in a liquid phase in the vessel by rotating the reaction vessel about its axis at a speed of 10 to 100 rpm, while said reaction vessel is kept inclined at an angle between 5.degree. and 45.degree. to the horizon with its mouth being raised; and an apparatus usable for practicing this method.

Abstract: The invention provides an immuno-reactant, being either an antigen or an antibody covalently bonded to a cross-linked film, more particularly composed of cross-linked protein or peptides, enveloping a carrier body, e.g. a solid, non-porous glass bead of 6 mm diameter. The immuno-logically active parts of the immuno-reactant are present on the surface in a form in which they are available for immunosorption. If the immuno-reactant is a protein, e.g. an antibody, it can provide all or part of the film-forming material. In a particular embodiment the film carries antibodies against a second type of antibody which is captured immunosorptively to form a "double layer" carrier-bound immunosorbent, the antigen capturing sites of the second type of antibody providing the immunosorption sites of the product. The immunosorbents are used, for example, for RIA or ELISA assays.

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: 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: A method for the detection and/or determination of an antigen having at least two separate antibody combining sites or antigenic determinants (i.e. a polyvalent antigen) using at least two different monoclonal antibodies which bind to the separate sites is described. In particular the method utilizes Protein A with a carrier to immobilize a first monoclonal antibody which in turn, can bind to one antigenic determinant on the antigen. The second antibody with a label is provided in a solution and binds to the second antigenic determinant on the antigen. A test kit is described for practicing the method. Novel anti-PGH synthase and anti-PGI.sub.2 synthase antibodies and hybridoma cells producing such antibodies are described.

Abstract: Colorectal carcinoma is detected by testing body fluids for the colorectal carcinoma monosialoganglioside identified by monoclonal antibodies produced by fused cell hybrid ATCC HB 8059.

Abstract: An improvement in assaying methods involving biospecific affinity reactions, in which there are used from 2 to 4 reactants, one of which, reactant (I), is labelled with at least one analytically indicatable atom or group and is soluble in the aqueous liquid in which the biospecific affinity reaction is carried out, the reactants forming, by means of biospecific reactions, a conjugate in which labelled reactant (I) is incorporated; and in which assaying methods the analytically indicatable atom or group is assayed in the conjugate and/or in labelled reactant (I), which is not bound to the conjugate. The conjugate that has been formed or labelled reactant (I) not bound to the conjugate is bound covalently to an insoluble carrier or to an insolubilizable carrier, which latter carrier is made insoluble after the covalent binding has been carried out, whereafter the assay of the analytically indicatable atom or group is carried out.

Abstract: In the immunoassay of a particular protein in a biological fluid, there is frequently interference in the assay by other proteins present in the fluid, e.g. by complement factors or antibodies in human serum. The interference so caused can be avoided by subjecting the fluid to protein-digestion, using for example an enzyme such as pepsin, as a result of which the particular protein of interest, or a fragment thereof, can be assayed without interference by the other proteins. Also, radioallergosorbent tests for particular IgE antibodies can be improved in sensitivity and accuracy, by subjecting the absorbed IgE to enzymic digestion, and then assaying a fragment thereof.

Abstract: Disclosed is a technique for screening populations to detect potential bladder cancer patients. The screening test is based on a discovered correlation between the respective ratios of C-reactive protein to total protein in urine and serum and the incidence of bladder cancer.

Abstract: A prostate antigen distinct from prostatic acid phosphatase has been detected in normal, benign hypertrophic and malignant prostatic tissues, but not in other human tissues. The prostate antigen was purified to homogeneity from prostatic tissues by ammonium sulfate precipitation, DEAE-BioGel A anion exchange chromatography, molecular sievings on Sephadex G-100 and Sephadex G-75, and preparative polyacrylamide gel electrophoresis. The purified prostate antigen shows a single protein band on analytical polyacrylamide gel electrophoresis and isoelectric focusing. The molecular weight of purified antigen was estimated by Sephadex G-75 gel filtration to be 33,000 and by sodium dodecyl sulfate polyacrylamide gel electrophoresis to be 34,000 with no subunit. The prostate antigen had an isoelectric point of 6.9.

Abstract: A homogenous sample of identical bispecific antibody determinants, each determinant being composed of two L-H half-molecules linked by disulfide bonds, each L-H half-molecule being specific for a different antigenic determinant and including at least the F(ab') portion of a monoclonal IgG antibody. The bispecific antibody determinants are useful, e.g., in the formation of multilamellar assemblies and enzyme electrodes.

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: Process for carrying out an analytical determination of the presence of a substance by means of chemiluminescence, comprising employing fluorescein isothiocyanate (FITC) as a labelling agent, triggering a chemiluminescence reaction by adding an aqueous solution of sodium hypochlorite, and measuring the emission of light.

Abstract: An immunological substance detecting reagent is provided which, in the preferred embodiment, employs an immunological homolog specific for the immunological substance to be detected coupled to a water-soluble polymer having a net charge not greater than zero. The water-soluble polymer further has associated a plurality of marker substances such as fluorophore molecules thereby providing a reagent of increased sensitivity for the detection of an antigen-antibody immunological reaction.

Abstract: Process for the quantitative and qualitative determination of antigens, antibodies and their complexes by means of a chemiluminescing labelling substance activated or excited to chemiluminescence by an analytical reagent. By means of a serological reaction, initially an antigen/antibody complex is formed which is treated with a chemiluminescing conjugate containing chemiluminescing triphenylmethane dyes and the chemiluminescence of the chemiluminescing complex formed is measured. Qualitative or quantitative details on the substances to be measured are obtained on the basis of the resulting measured values. The dye labels are activated by either hypochlorites or mixtures of H.sub.2 O.sub.2 and a chloramine.

Abstract: An improved method for the preparation of .sup.125 I-labelled Protein A (.sup.125 I PA) of high specific and functional activity. .sup.125 I PA has been used in combination with purified rabbit IgG (immunoglobulin G) bound to a solid support to develop a competitive binding assay capable of detecting Protein A or human, rabbit and guinea pig IgG at the nanogram level. Additionally, .sup.125 I PA may be used to detect methotrexate, leucovorin and similar substances..sup.125 I PA has also been used to detect IgG anti-Forssman antibody bound to sheep erythrocytes and to line-1 and line-10 tumor cells and as an indirect assay for tumor associated antigen in the ascitic fluid of tumor-bearing guinea pigs.Additionally, an improved method of preparation of iodination of Protein A is utilized. This procedure used the Bolton-Hunter (1973) reagent of radioactive iodine in benzene which carrier is evaporated.

Abstract: A reaction vessel, test device, and method of detection or measurement are disclosed, featuring the use of at least two operatively connected zones formed by transport surfaces spaced apart throughout most of the zones a capillary distance. The zones are fluidly connected by meniscus control means effective to stop capillary flow of the liquid from one zone to the other, until an externally generated actuation pressure is applied.

Abstract: A fluid receptacle is provided with a retainer on the inner surface thereof for confining at least the reactive portion of a coated insert matrix within the receptacle during inversion thereof. The insert has a configuration permitting fluid to be poured into the receptacle and decanted from the receptacle by inversion of the receptacle with the reactive portion of the insert confined therein. The various decanting, washing and measuring operations involved in an assay may therefore be performed while the insert remains confined within the original receptacle, eliminating the need for additional receptacles and for manual assembly or disassembly operations during the assay.

Abstract: A monoclonal antibody is described characterized by its specificity to interferon-.alpha. (leukocyke interferon). Preferably the antibody has specificity to human interferon-.alpha.. The monoclonal antibody is secreted by a hybrid cell formed by the fusion of lymphocyte cells derived from an animal immunized with interferon-.alpha., with mycloma cells. The antibody has applications in the purification of interferon. It may be covalently attached to a solid support and used as an immunoadsorbent. Purifications of up to 5000 fold may be obtained in a single pass through an immunoadsorption column of this type. An immunoradiometric assay for interferon-.alpha. is also described.

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: Process for chemically binding organic compounds containing carbohydrate residues, onto a support bearing at least one reactive --NH.sub.2, in which at least one --CH.sub.2 OH group of the carbohydrate residue is transformed in a --CHO group, by oxidation and then the --CHO groups thus obtained are reacted with at least a reactive --NH.sub.2 carried by the side chains covalently bound on a solid, insoluble support, the side chains are chosen from among amines, polyamines, diacids, amino-acids, hydrazines, and are eventually coupled, by the intermediary of their reactive --NH.sub.2, with a nitrogen-containing compound chosen from aliphatic or aromatic amines, aliphatic or aromatic hydrazines, or amino acids, comprising eventually jointly a --SH group and a --NH.sub.2 group.Products resulting from this process and biological reagents containing said products as their active constituents.

Abstract: An improved immunoassay sample determination process for determining the presence of a component of an antigen-antibody reaction in a sample is disclosed which substantially eliminates non-specific interactions between the sample and the reaction vessel wall surfaces during the antigen-antibody reaction, to thereby greatly increase the accuracy of the process. In practice, a site-deactivating medium such as an animal- or vegetable-derived total biological fluid or extract (e.g., plasma or gelatin) is covalently bound to the vessel wall surfaces for deactivation purposes. In preferred forms the process is solid-state, wherein one of the components of the antigen-antibody reaction is coupled covalently to the coating medium, and a reaction mixture fraction including the sample being determined and the other component which has been labeled, for instance by means of a colorimetrically active enzyme.

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: The invention relates to a process for the determination of serum lipoproteins by an immuno-enzymatic method. The process of the invention consists in reacting a specific antibody of the apoprotein whose amount is to be measured, fixed on a support, with, on the one hand, the compound for coupling the corresponding lipoprotein with an enzyme and, on the other hand, the analyzed sample. The amount of enzyme fixed is then an inverse function of the apoprotein content of the sample. The support with the products which are fixed thereto are separated from the rest of the reagents and the activity of the enzyme of the support is measured which is compared with standard measurements.The process is useful for serum analyses, particularly for early detection of cardiovascular disease risks.

Abstract: Process for the preparation of test reagents comprising antigens or antibodies adsorbed on a surface, for example, the surface of synthetic or natural polymer particles in which the test material to be adsorbed is dissolved in a solvent in contact with the adsorbing surface and precipitated by the addition of a liquid which is miscible with the solvent, but does not dissolve the test material.

Abstract: A multilayer analytical element for detecting a ligand in or the ligand binding capacity of a liquid sample of the type having a reagent layer incorporating reagents which are responsive to the ligand in or the ligand binding capacity of the sample to give a detectable response, a radiation diffusing and blocking layer, and a support layer, the improvement wherein the radiation diffusing and blocking layer is (a) interposed between the reagent layer and the support layer; (b) impermeable to the ligand, reagents of the reagent layer, and products of their interreaction; and (c) inert to the ligand, reagents of the reagent layer, and products of their interreaction.

Abstract: Immunoassays are performed with two different ligands tagged with two different tagging constituents which are independent of each other and the two tagged ligands are immunologically bound together. The two different ligands may be detected independently through their independent tagging constituents for quality control, internal calibration (standardization), determination of viability and shelf life and the like.

Abstract: A method for indirect measurement of the concentration of a free ligand in a liquid sample also containing the ligand bound to endogenous binding material, comprising the steps of performing two separate measurements, wherein the first measurement is used to determine the total concentration of free and reversibly bound ligand in the sample to be tested, and the second measurement comprises the steps of (i) exposing separately to a known amount of a labelled ligand (a) the sample to be tested, and (b) each of a plurality of standard solutions containing known concentrations of the free ligand, (ii) thereafter exposing each resulting mixture from (i) (a) and (i) (b) separately to an immobilized binding agent to bind a portion of the free labelled ligand and a similar proportion of the free unlabelled ligand to the immobilized binding agent in each mixture, (iii) separating the immobilized binding agent with its bound ligand from the residual material in each mixture, (iv) measuring the proportion of labelled l

Abstract: A heterogeneous specific binding assay which employs a substance having reactant activity, i.e., a reactant, in a chemiluminescent reaction 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 chemiluminescent reactant. The activity of the conjugated reactant in the chemiluminescent reaction, i.e., the production of light, is utilized as means for monitoring the extent of binding of the labeled constituent in conventional heterogeneous specific binding assay schemes. The presence of a ligand in a liquid medium may be determined following conventional competitive binding manipulative techniques.

Abstract: A highly sensitive, immunoassay method for determining the amount of an analyte in a sample containing a known analyte in an unknown concentration is provided. Sample; a polypeptide-labeled analog of the analyte, an antibody specific for said analyte, a polypeptide partner capable of non-covalently binding with the polypeptide-labeled analyte to form a complex having catalytic activity, and a substrate capable of being converted to a reporter molecule by the catalytic activity of said complex are brought together in a medium. The polypeptide-labeled analyte analog is capable of competitively binding to the antibody and the polypeptide partner, the antibody inhibiting the formation of a catalytically active complex in the absence of analyte, and the concentrations of the antibody, polypeptide partner and polypeptide-labeled analyte are such as to cause varying amounts of analyte to be directly related to the conversion of the substrate to the reporter molecule.