Abstract: Assay methods are provided for determining an analyte in a sample suspected of containing the analyte. The method is carried out using a composition that includes a conjugate of a first sbp member with a particle. A luminescer is reversibly associated with a nonaqueous phase of the particle. Where the first sbp member is not complementary to the analyte, a second sbp member that is capable of binding to the first sbp member is employed. Unbound conjugate is separated from conjugate that is bound to the analyte or to the second sbp member. A reagent for enhancing the detectability of the luminescer is added and the light emission of the luminescer acted on by the reagent is measured.

Abstract: The present disclosure is directed to methods for the preparation of immunoadsorption matrices having IgG molecules adsorbed thereto in a preferred configuration, i.e., adsorbed to the matrix by their (Fc) rather than F(ab) portions. IgG molecules, are selected such that the F(ab) portion of the IgG fraction adsorbed has a more acidic or basic net isoelectric point or pI range than the F(c) end of the molecule, depending on the characteristics of the adsorption surface. For negatively charged surfaces, IgG molecules having relatively alkaline F(c) portions are selected. For positively charged surfaces, IgG with relatively acidic F(c) portions are selected. Additional selection criteria include pI fractionation to provide fractions having well defined pI characteristics as defined by "non-overlap" or "pI range" of F(c) and F(ab) portions pI's. Methods disclosed are particularly well suited to the preparation of colloidal gold immunostains.

Abstract: An immunoadsorbent material for removing IgM and IgM-complexes from biological fluids is prepared by covalently binding anti-IgM antibodies to a solid-phase silica matrix. It has been found that reacting hydroxy-derivatized silica in the presence of cyanogen bromide with the anti-IgM antibodies provides a particularly stable, high-capacity immunoadsorbent. The immunoadsorbent material may be employed in a column for therapeutic treatment of various disorders, such as primary biliary cirrhosis.

Abstract: This invention provides a means for determining the concentration of any of a wide range of antibody or antigen molecules with a high degree of specificity, accuracy and sensitivity. Antigen or antibody concentration is determined by effecting an agglutination reaction in a liquid medium and determining the cluster size distribution of agglutinated particles by optical pulse particle size analysis. The measured cluster size distribution then is compared with a standard quantitative relationship between the cluster size distribution and concentration of the antigen or antibody being tested. By this means one may specifically ascertain the absolute concentration of the antigen or antibody in question in the sample being analyzed. In addition to detecting antigen or antibody molecules, the process of this invention can be used to determine the concentration of any substance capable of specifically promoting or inhibiting an agglutination reaction such as viruses, white blood cells or the like.

Abstract: An immunoassay for detecting and measuring hCG in a sample includes an antibody directed to the carboxy terminal portion of the .beta. subunit of hCG and a monoclonal antibody directed to a determinant on hCG at a locus sufficiently remote from the carboxy terminal portion of the .beta. subunit of hCG that both antibodies can simultaneously bind to hCG, wherein at least one of the antibodies is delectable when both are bound to hCG.In a presently preferred embodiment, an immunoassay for hCG or hCB.beta. in urine includes a purified, labeled or detectable serum-derived antibody directed to the carboxy-terminal portion of the .beta. subunit of hCG and a matrix-bound monoclonal antibody directed to a locus on the .beta. subunit sufficiently remote from the carboxy-terminal portion that both antibodies can simultaneously bind to hCG or hCG.beta..

Abstract: An improvement in a method to conjugate a protein which must resist denaturation with a variable component using a carbodiimide condensing agent utilizes a polar aprotic solvent as the medium for condensation. This improvement results in shorter reaction times and improved coupling efficiency.

Abstract: New method for the detection of specific binding agents and their corresponding bindable substances by employing a label which is a latex particle which can be visually detected.

Abstract: A competitive immunoassay for a steroid receptor has been developed in which monoclonal antibodies capable of binding to the steroid receptor are competitively bound by anti-steroid antibodies capable of binding to the steroid. The presence or amount of monoclonal antibody-anti-steroid antibody complex formed is related to the amount of steroid receptor present in the assayed material.A histochemical assay is also provided for detecting the amount of a steroid-receptor complex in a biological sample. This assay involves (1) adding an amount of steroid to the sample to form a steroid-receptor complex; (2) contacting the complex with a monoclonal antibody capable of binding to the complex; (3) removing any unbound monoclonal antibody; (4) adding a detectably labeled antibody capable of binding to the monoclonal antibody; (5) determining the amount of labeled antibody bound to the monoclonal antibody; and (6) determining the amount of steroid-receptor complex.

Abstract: The invention relates to a method of immunoassay for monoamines (molecules having a primary or secondary amine function) comprising chemical quantitative conversion of such amines into derivatives of higher molecular weight, which thereafter are brought into competition with radioactive analogous, or analogous carrying a tracer, for fixation to an antibody capable of recognizing all of them.

Abstract: A method is disclosed for determining an analyte in a sample suspected of containing the analyte. The analyte is a member of a specific binding pair (sbp) consisting of ligand and complementary receptor. The method comprises combining in an aqueous medium (1) said sample, (2) a first reagent comprising a non-dispersed surface to which is bound an sbp member that becomes bound to the surface in relation to the amount of analyte in the sample. The volume of the aqueous medium containing the above reagents is sufficiently large to allow complete immersion of the first reagent therein and determination of the analyte but not so large to result in substantial interference in the determination upon addition of a second reagent reactive with the conjugate of enzyme and sbp member and capable of generating a signal. The second reagent is present in a medium of sufficient volume to substantially increase the volume of the liquid medium and reduce interference in the determination.

Abstract: Device for the carrying out of an immunochemical determination comprising at least one holder which is provided with an opening at the top and to the inside of which an immunochemically active substance has been applied. The device is closed at the top by a removable closure means and the holder contains at least a second immunochemically active substance which, in the absence of test medium, in the form in which the second immunochemically active substance is contained in the closed holder, during storage and transport under normal conditions exhibits no interaction with the inside of the holder and is inert with respect to the immunochemically active substance applied thereto. The holder contains a solid, more or less spherical, freeze-dried particle which contains the second immunochemically active substance.

Abstract: A solid phase immunoassay comprises the steps of(a) immobilizing an immunoreagent on the surface of a carrier comprised of an inert synthetic resin selected from the group consisting of polyimides and polyfluorinated synthetic resins,(b) contacting the immunoreagent with a complementary immunoreagent whereby an immunocomplex immobilized on said carrier is formed,(c) quantitating the immobilized immunocomplex.An element useful in conducting this solid phase immunoassay is prepared by a process of treating the surface of an article comprised of a synthetic polymer selected from the group consisting of polyimides and polyfluorinated synthetic resins to make it adsorptive of an immunoreagent which comprises the steps of(a) thoroughly rinsing the surface with a water-miscible organic solvent,(b) thoroughly rinsing the surface with water.

Abstract: The method of measuring circulating antigens or antibodies by using a ligand labeled specific antigen or ligand labeled specific antibody chemically attached to a soluble matrix or backbone, a differently labeled anti-antigen or anti-antibody and a solid phase anti-ligand directed at the ligand attached to the specific antigen or specific antibody. This is achieved by either one or two analytical schemes:(a) Reacting a patient sample with a ligand labeled specific antigen or a ligand labeled specific antibody in the liquid phase in the presence of a differently labeled specific anti-antigen or labeled specific anti-antibody. This immunological complex is reacted with an immobilized anti-ligand on a solid support which is directed against the ligand attached to the specific antigen or antibody through the liquid matrix. Subsequently the solid phase is washed and checked for the label on the anti-antigen or anti-antibody which is directly proportional to the concentration of specific antigen or antibody.

Abstract: A digoxin derivative/immunogenic protein conjugate is disclosed which has the carbohydrate moiety of digoxin intact. Antibodies raised against this conjugate show minimal cross-reactivity to digoxin metabolites enabling the use as an antibody in the diagnostic analysis for digoxin when measured in the presence of its metabolites found in serum isolated from a human.

Abstract: Divalent hapten derivatives wherein two hapten moieties are connected by means of a bifunctional spacer wherein the derivative has the formulaA--X--Awhere A is a bonded hapten moiety and X is a bifunctional spacer having the formula(B).sub.m --Y--(CH.sub.2).sub.n --Z--(CH.sub.2).sub.n --Y--(B).sub.mwhere m is independently 0 or 1, B is (CH.sub.2).sub.n', wherein n' is an integer from 1 to 4, or CO(CH.sub.2).sub.n", wherein n" is an integer from 2 to 4; Y is independently --CONH--, NHCO--, OOC--, --COO--, --O--, --S--, or --NR--, wherein R is hydrogen or alkyl; n is an integer from 1 to 10; and Z is an organic moiety containing at least one hydrophilic atom are disclosed.

Abstract: A first glycoprotein having a molecular weight of approximately 61,000-68,000 daltons in the MJ, C5-MJ, C91 PL or HUT-102 cell lines, of which 46,000 to 48,000 is the unglycosylated moiety, is obtained from cells infected with human T cell leukemia virus. A second glycoprotein having a molecular weight of approximately 45,000-52,000 daltons is also obtained from such cells and is in large part identical to the NH.sub.2 -terminal end of the first glycoprotein. The presence, in a biological specimen, of antibody to the antigenic determinant of either of these proteins is indicative of the presence of cells infected by human T cell leukemia virus. An assay for the antibody is a useful diagnostic procedure for determining such infection in biologial specimens.

Abstract: Detection of an identified human carcinoma tumor antigen in a pathological sample by means of a labelled monoclonal antibody specific to the determinant site on the antigen is enhanced and/or accelerated at an earlier development stage than heretofore achieved by removing a carbohydrate steric hindrance for monoclonal antibody availability to bind the antigen of the tumor for which it is specific. The carbohydrate steric hindrance for monoclonal binding to the antigen is identified as sialic acid. The method of the invention involves selective removal of sialic acid from the antigen's determinant site by enzymatic digestion using neuraminidase.

Abstract: A method of identifying the presence or absence of an infected state in the living body by obtaining a biological fluid sample from a living body, adding to the sample a material known to bind with immune complex, adding to samples of the resulting material labelled reagents known to react specifically with antigen portions of different immune complexes, separating excess of the reagents and detecting the presence or absence of the label in the residual mixtures thereby to identify the nature of any reacted antigen portion of immune complex therein.

Abstract: A process for producing an antibody having a high specificity to a first antigen and a low cross-reactivity with at least one second antigen, said first antigen comprising a desired antigenic determinant and said second antigen comprising at least one antigenic determinant which is structurally related to said desired antigenic determinant of said first antigen, which process comprises treating a mammal with a copolymer of D-glutamic acid and D-lysine coupled with said second antigen whereby to induce a substantially effective immunological tolerance to said second antigen and then immunizing said mammal with said first antigen. This process results in a higher productivity of a mammal cell capable of producing said desired antibody by culturing said mammal cell, for example, by forming a hybridoma with a suitable tumor cell and implanting the hybridoma into another mammal. The antibody or antiserum of this invention may with advantage be used for immunoassay such as radioimmunoassay.

Abstract: Novel particle reagent for light scattering immunoassays are provided. The particle reagents are high refractive index shell-core polymers, having at least a partial surface coverage by a monomolecular layer of anionic surfactant, covalently bonded to compounds of biological interest. The novel particle reagents are particularly suited to protein immobilization by covalent bonding to the shell and are especially useful for light scattering immunoassays.

Abstract: A method of measuring the result of a binding assay that does not require separation of fluorescent smaller particles is disclosed. In a competitive binding assay the smaller fluorescent particles coated with antigen compete with antigen in the sample being analyzed for available binding sites on larger particles. In a sandwich assay, the smaller, fluorescent spheres coated with antibody attach themselves to molecules containing antigen that are attached to larger spheres coated with the same antibody. The separation of unattached, fluorescent smaller particles is made unnecessary by only counting the fluorescent events triggered by the laser of a flow cytometer when the event is caused by a particle with a light scatter measurement within a certain range corresponding to the presence of larger particles.

Abstract: A preparation for the detection of chlamydial infections using lipopolysaccharide of Re-lipopolysaccharide mutants of gram-negative bacteria. The lipopolysaccharide preparation is used in the production of group-specific antibodies to chlamydiae for diagnostic purposes or for the demonstration of antibodies to chlamydial group antigen in specimens.

Abstract: Microcapsules for effecting an immune response, especially through agglutination reaction, comprise an antigen or antibody bound to functional groups, such as amino groups, carboxy groups, etc., on a wall surface of the microcapsules via cross linking agents, such as polyfunctional isocyanates, isothiocyanates, etc. The antigen or antibody is strongly bound to the wall surface of the microcapsules to thereby achieve excellent detection sensitivity.

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: An immunoassay is provided for cholesterol epoxide. To prepare the antibodies used in the immunoassay, novel immunogens, are prepared which comprise a 3,5(6)-transdiaxial-dihydroxycholestane-6(5)-yl-hapten adduct linked to a covalently bonded bridge to a carrier protein. To detect cholesterol epoxide in the sample, it is converted to the hapten adduct, then contacted with the selected antibody.

Abstract: Chemically synthesized polypeptides containing about 6 to about 40 amino acid residues having amino acid residue sequences that substantially correspond to the amino acid residue sequences of antigenic determinants of interleukin-2, when administered alone or as polymers or as conjugates bound to carriers, induce the production of antibodies of predetermined specificities. The polypeptides and the antibodies produced thereto can be used in diagnostic systems to measure the presence and amount of interleukin-2 and interleukin-2 cell surface receptors or binding sites in an assayed sample.

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: 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 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: HBcAb in a biological fluid is adsorbed on a surface which is then coated with BSA. The coated surface is then incubated first in the sample and then in the presence of radiolabelled HBcAb.

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: Chloramphenicol derivatives are provided for use in the preparing of antigen conjugates for the production of antibodies specifically for chloramphenicol. Specifically, the aryl amino group is derivatized to introduce a non-oxo-carbonyl group which is used for amide formation with an antigen. The conjugate is then injected into a vertebrate for production of antisera which is isolated in conventional ways and find particular use in competitive protein binding assays.

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: A composition having a protein binding solid support onto which is bound a mixture of antigens and antibodies which are both bound to the solid support individually and are not present in the form of an immune complex.

Abstract: Competitive assay procedure for determining the concentration of physiologically active haptens in which the hapten is bound to an immunogenic carrier and competes for binding sites on antibodies against the hapten. Actual concentration is determined by comparison with standard curves.

Abstract: A method for detecting chronic myelogenous leukemia in a human comprising the step of testing a biological sample from a patient to determine the presence of a marker protein (P210) which is characterized as a c-abl protein having tyrosine kinase activity and a molecular weight of approximately 210,000. Antisera which are specific for the P210 protein are disclosed which can be used to precipitate the P210 protein to allow identification by gel electrophoresis or other technique.

Abstract: Conjugates of penicilloic acid derivatives and certain poly(amino acids), which are either antigenic or enzymatic, are provided. Antibodies raised against the antigenic poly(amino acids) and the enzyme conjugates are used as reagents in immunoassays. In particular, the compounds of the present invention can be used for measuring the presence of a .beta.-lactamase in a patient serum sample by adding a known amount of penicillin to the sample and observing the production of penicilloic acid over a fixed period of time.

Abstract: Method and test kit for detecting Clq-containing complexes in human serum containing native serum Cl. The method uses a monoclonal antibody which selectively reacts with human Clq in the presence of native human serum Cl. Preparation of hybridomas generating such antibodies is also disclosed. The method is applicable to detection of autoimmune diseases and AIDS.

Abstract: The present invention concerns an autologous precipitating antibody and the gp70 pigment-associated antigen on melanoma cells which it recognizes. The antibody is useful in detecting pigmented melanoma cells in excised specimen, serum or urine.

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: A suspension of diagnostic particles comprising antibody molecules attached to a carboxylate derivatized polymer core is provided for agglutination tests. The antibody is linked to the core through an avidin-biotin bridge. Avidin is joined by an amide bond to carboxyl groups on the core, and biotin is linked by an amide bond to amino groups on the antibody molecule. The core-bound antibody is exposed to a mixture of free biotin and biotinylated antibody to attach a controlled amount of antibody that is consistent with suspension stability prior to its use in a test and rapid cross-linking of suspended particles in the presence of antigen.

Abstract: Particle reagents, having polyether polyamine linking groups, for use in turbidimetric immunoassays are provided. These linking agents permit ready covalent attachment of compounds of biological interest to polymer particles and lead to immunoassays of improved precision.

Abstract: Compositions, articles of manufacture, and methods are provided for enhanced binding of compounds to a surface. The compositions find particular use in coating wells, slides, other surfaces, where ligands or receptors are to be bound and detrimental non-specific binding may be encountered. Particularly, proteins are modified by alkylation to produce a product which strongly adheres to a surface, allows ligands and receptors to be firmly attached by means of the alkylated protein to a surface under mild non-denaturing conditions and permits procedures to minimize non-specific binding of proteins without significant loss of the materials of interest.

Abstract: Enhanced immunogenicity is achieved by covalently linking immunogens to liposomes and injecting the membrane-bound-proteins into an appropriate vertebrate host. Methods and compositions are provided for producing antibodies.

Abstract: This invention relates to a kit for the detection of microbial nucleic acids and a method for identifying the nucleic acids using a one-step sandwich hybridization technique. The technique requires two complementary nucleic acid reagents for each microbe or group of microbes to be identified.

Abstract: The present invention concerns novel immunoprecipitating autologous antibodies which recognize the Class 1 gp90 antigen on melanoma cells. These antibodies, optionally tagged with a chromophoric or radioactive label and immobilized on an inert support, may be used to recognize and isolate the gp90 antigen from melanoma cell extracts. Monoclonal antibodies to melanoma may be screened with the gp90 antigen for those which recognize epitopes other than the FD antigenic system.The cell line containing the gp90 antigen which has been cultured in vitro is a source of gp90 antigen for generation of monoclonal antibodies which will be useful in analyzing the gp90 antigen for those epitopes which may be of diagnostic value in immunoassay of melanoma.

Abstract: A carrier composed of particulate polymer of at least one monomer represented by the general formula: ##STR1## wherein R.sup.1 and R.sup.2, which may be the same or different, are hydrogen atoms or alkyl groups having 1 to 4 carbon atoms; R.sup.3 is a straight-chain or branched alkylene group having 2 or 3 carbon atoms, which group may be substituted by a hydroxyl group; X's, which may be the same or different, are halogen atoms; m is zero or an integer of 1 to 10; and n is an integer of 1 to 5, or a particulate polymer of said monomer and at least one other copolymerizable monomer. Said carrier is suitable for supporting a biological substance such as an immunoreactive substance, an enzyme, a cell, a cell-discriminating substance, and the like, particularly an immunoreactive substance.

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: A heterogeneous immunoassay for digoxin. An excess of labeled anti-digoxin antibody is added to a test sample and the resulting reaction mixture is contacted with a solid phase having ouabain immobilized thereon. All the free, labeled antibody binds to the oubain. The eluted digoxin anti-digoxin antibody complex is measured for label activity. A competitive mode is also disclosed.

Abstract: Immunological preparations are prepared by immunizing hens with an immunogen having a molecular or particle weight of at least 30,000, to a stage of hyper-immunization at which there occurs a plateau-like levelling-off of the antibody content of the serum. The immunogenicity of the immunogen can be enhanced by enlarging the immunogen particle mass. The eggs of the immunized hens are collected, the yolk is separated from the eggs, followed by separation of the lipid content of the yolk. The antibodies in the egg yolk are then rendered indispersable with the aid of a water-soluble linear filamentary non-charged polymer precipitant such as PEG and the indispersable antibodies are recovered. This precipitation of antibodies is advantageously preceded by a precipitation of caseinaceous proteins, lipid and yolk particles at lower polymer concentrations. Selected antibodies (IgY or IgG) can be concentrated by pH-controlled fractional precipitation with PEG.