Abstract: This invention relates to a flotation immunoassay employing a novel buoyant matrix to which an antigen or antibody is coupled and which separates the bound and free products of the assay by floating to the surface of the reaction liquid. The novel flotation device which makes it possible to detect and to quantitate either antigen or antibody can also be used to fractionate cells and molecules.

Abstract: The sequence of the protein coding regions of the bcl-2 gene are provided as well as bacterial clones which produce the proteins. Assays are provided for detecting a class of B-cell neoplasms associated with a chromosome translocation between chromosomes 14 and 18. This translocation is involved in the majority of cases of human follicular lymphomas. One assay employs an antibody which is immunoreactive with a human protein which is over-expressed due to the chromosome translocation. Another assay involves measurement of the amount of mRNA which hybridizes to the gene proximal to the translocation break-point.

Abstract: Disclosed are immunological methods and materials for detection of antigens associated with breast or prostate cancer disease states. Presently preferred antibody preparations (e.g., PR92 monoclonal antibodies produced by hybridoma cell line ATCC HB 9390) are employed in immunoassays performed on patient body fluids and for purification of tumor-associated antigen compositions.

Abstract: A two site cross-reaction immunometric sandwich assay method for the detection and measurement of an analyte, such as creatine phospho-kinase-MB, in serum comprising the selection of two different antibodies each of which is specific to a different analyte but each of which will cross-react with the analyte of interest. The first antibody is reacted with the unknown sample utilizing a solid-phase to bind the first antibody. Separation of the solid and liquid portions of the first reaction is accomplished and the solid portion thereof is reacted with the second antibody which is tagged. The solid portion and liquid portion of the second reaction are separated and the solid portion is tested for the tag as an indication of the presence of said analyte. With particular reference to testing for creatine phospho-kinase-MB in human serum, the cross-reacting antibodies utilized are antibody to creatine phospho-kinase-BB and creatine phospho-kinase-MM.

Abstract: A two site cross-reaction immunometric sandwich assay method for the detection and measurement of an analyte, such as creatine phospho-kinase-MB, in serum comprising the selection of two different antibodies each of which is specific to a different analyte but each of which will cross-react with the analyte of interest. The first antibody is reacted with the unknown sample utilizing a solid-phase to bind the first antibody. Separation of the solid and liquid portions of the first reaction is accomplished and the solid portion thereof is reacted with the second antibody which is tagged. The solid portion and liquid portion of the second reaction are separated and the solid portion is tested for the tag as an indication of the presence of said analyte. With particular reference to testiong for creatine phospho-kinase-MB in human serum, the cross-reacting antibodies utilized are antibody to creatine phospho-kinase-BB and creatine phospho-kinase-MM.

Abstract: The method described coats the interior of a tube-like support with the binding partner of the analyte. Analyte and anti-analyte-label conjugate are added. When the tube-like support is rotated end-over-end, the thin layer of analyte and conjugate readily react with the binding partners to form an immobilized label complex. When rotation stops, the reaction quenches to provide precise timing. The label complex is released and measured.

Abstract: Compositions and methods are disclosed for multiple simultaneous assays of different analytes using radioactive labeled antibodies to the analytes, at least one portion of the assay being an immunoradiometric assay in which there is employed a metal isotope label, e.g., .sup.57 Co, attached to an antibody to the analyte through a chelator, e.g., ethylenediaminetetraacetic acid. Multiple simultaneous immunoradiometric assays can be performed by this method, as can multiple simultaneous assays in which one portion of the assay is an immunoradiometric assay and another portion or portions involve one or more other radioassay techniques.

Abstract: Incubation media intended for solid-phase immunometric assays and containing lactoferrin are described. Compared with the media used hitherto, these incubation media have the advantage that they prevent, especially in the case where heated sample material is used in the assays, false results on determination of antigens or antibodies.

Abstract: A sandwich immunoassay for determining total monoclonal IgG content in samples of purified IgG, cell culture medium and ascites fluid is provided based on the utilization of anti-light-chain IgG antibodies as both capture and label antibodies.

Abstract: A method for analyzing antigen or antibody contained in a sample by introducing a first end surface of at least one measuring light guide member and a second end surface of at least one reference light guide member in the sample. Each of said measuring and reference light guide members being coated with a protection layer except for said first and second end surfaces and an other end surface of respect light guides. Only on said first end surface of the measuring light guide member being fixed an antigen or antibody which is specifically reactive with the antigen or antibody contained in the sample to effect an antigen-antibody reaction. Photoelectrically detecting optical properties of said first and second end surfaces of the measuring and reference light guide members, respectively, by transmitting light through the meauring and reference light guide members to derive detection signals as a measure of antigen or antibody in the sample.

Abstract: An improved heterogeneous immunoassay is provided which includes the utilization of support materials capable of reversible immobilization of proteinaceous binding partners of biological material of interest and the removal of labeled complexes from these supports through the use of release reagents permitting the subsequent solution phase determination of the amount of label.

Abstract: Gas containing granules of a resiliently compressible material having a density variable with pressure are used as a solid phase support in liquid phase reactions such in immunological reactions. Varying pressure causes the density of the granules to change and is used for mixing and separating the granules in liquid phase. In an immunological reaction, the granules having an attached antibody are combined with a solution containing an antigen, then varying pressure is applied to the solution to mix the granules in the solution to fix the antigen to the antibody and then pressure on the solution is varied to force the granules to the top or bottom of the solution whereby the solution can be separated from the granules.

Abstract: Methods are disclosed for detecting an antigen in a biological sample. The methods involve providing in combination a solid support, which is substantially free of specific binding proteins, and a medium comprising an antigen from the sample and an antibody for the antigen. The combination is incubated under conditions sufficient for the antibody when bound to the antigen to bind to the support. The presence or amount of antibody on the support or in the medium is determined and is related to the presence of antigen in the sample. The methods have particular application to the detection of gram-negative bacteria.

Abstract: Preparation of novel polypeptide sequences spanning amino acid position 61 of the ras protein, said sequences being characterized as containing a leucine in that position instead of the glutamine found in normal ras protein, the utilization of such polypeptides to prepare immunogen compositions utilizing such polypeptide covalently linked to immunogenic carrier materials, the production of antibodies elicited by such polypeptides, the screening of such antibodies to provide monoclonals specific to the P21Tleu61 transforming protein and to immunoassay and use in immunochemical methods employing such antibodies to determine the presence of the p21TLeu61 transforming protein.

Abstract: A secondary antibody capable of stabilizing the binding of a small molecule to its binding protein is described which secondary antibody is capable of binding said binding protein in the presence of an in the absence of the small molecule but is not capable of binding said small molecule in the absence of binding protein. Such antibodies may be obtained by forming a complex between a small molecule and its binding protein, using the complex to raise antibodies and selecting the antibodies. The antibodies may be used in competitive assays in which it is desired to improve the binding of a small molecule or labelled small molecule to its binding protein.

Abstract: An enzyme immunoassay based on membrane separation of antigen-antibody complexes wherein human or animal body fluid specimens containing an antigen are mixed with an enzyme-conjugated antibody specific for the antigen under test. Following incubation the antigen-antibody-conjugate mixture is passed through a filter membrane having an electrostatic charge providing an affinity for retaining antigen-antibody-conjugate complexes while not retaining free antibody-conjugate. Following washing of the filter membrane to remove free antibody-conjugate remaining thereon an enzyme substrate-chromogen reagent solution is applied to the filter membrane, which reacts with filter-bound antibody-conjugate and develops a visible or fluorogenic color indicative of the presence of antibody-conjugate.

Abstract: The present invention provides a process for the determination of an antibody by incubation with three different reagents R.sub.1, R.sub.2 and R.sub.3, of which R.sub.1 and R.sub.3 are present in liquid phase and are bindable with the antibody, R.sub.2 is present bound to a solid phase and is bindable with R.sub.1 and R.sub.3 carries a label, separation of the solid phase from the liquid phase and measurement of the label in the solid phase, wherein as R.sub.1 there is used a conjugate of a substance specifically recognized by the antibody to be determined and a reaction component of a specific binding system, as R.sub.3 there is used a conjugate of a substance specifically recognized by the antibody to be determined and a label and as R.sub.2 there is used the other binding component of the specific binding system.The present invention also provides a composition for the determination of an antibody, wherein it contains two different soluble reagents R.sub.1 and R.sub.

Abstract: A method for the determination of a substance present in a sample, which comprises:(a) contacting a sample containing a substance with:(i) a first immunological binding partner to the substance, wherein the first immunological binding partner is a monoclonal antibody bound to biotin or a biotin-binding protein,(ii) a second immunological binding partner to the substance, wherein the second immunological binding partner is a detectably labeled region specific polyclonal antibody, and(iii) a biotin binding protein or biotin bound to a carrier;(b) incubating the components of step (a) for a period of time and under conditions sufficient to form an immune complex between the substance, the first immunological binding partner, the second immunological binding partner, and the carrier;(c) separating the carrier from the sample; and(d) determining the detectably labeled second immunological binding partner in either the sample or the carrier;wherein the reaction between the first immunological binding partner and th

Abstract: A test system and procedure for quantitatively assaying biological material for a target immunological substance by means of immunochemical binding of immune complexes, comprising the target substance and its immunospecific conjugate, to insolubilized non-immunospecific factor, such as Clq. A sample of biological material suspected of containing the target substance is introduced into the test system including pre-determined amounts of the target substance and its immunospecific conjugate forming immune complexes having a known degree of chemical binding to the non-immunospecific factor. The amount of target substance present in the test sample is determined according to the deviation from the known degree of immunochemical binding caused by the addition of the sample to the test system, by reference to a standard curve.

Abstract: A method for determining the level of infectious bursal disease virus IBDV neutralizing antibody in poultry comprising a labeled monoclonal antibody R63 having the IBDV neutralizing capability of the monoclonal antibody expressed by hybridoma cell line ATCC HB-9490. Monoclonal antibody R63 is specific to all known IBDV strains and serotypes and competes only with itself and other antibodies recognizing the same neutralizing epitope present in poultry sera. In a competition assay, the increase in unbound labeled monoclonal antibody is an indication of the level of IBDV neutralizing antibody present in poultry sera.

Abstract: A method for enzyme immunoassay includes contacting under binding conditions a liquid suspected of containing an analyte, an antianalyte affixed to a solid support and a tracer having an enzyme conjugated thereto. A bound fraction is separated from the liquid and incubated in a second liquid with a masked ligand. The masked ligand is converted by the enzyme on the bound fraction to give free lignad which binds to an antiligand. A signal system, such as a signal enzyme and substrate therefor, or a label-loaded vesicle and vesicle lysing agent, is added to generate a signal used to detect or measure the analyte in the liquid. The invention includes a kit of materials useful in performing the assay of the invention.

Abstract: Methods and reagents for early detection of myocardial infarction determine the level of CK-MM.sub.A and the level of the combined CK-MM.sub.A and CK-MM.sub.B in a serum sample. From these measurements, the time of the acute phase can be more accurately determined. Novel anti-(CK-MM.sub.A) antibodies, anti-(CK-MM.sub.B) antibodies, anti-(CK-MM.sub.(A+B)) antibodies, labeled and insolubilized derivatives of these antibodies, labeled CK-MM isoforms, and kits containing one or more of these reagents are also described.

Abstract: An immunoassay for trichothecenes that have at least three hydroxyl groups at specified positions is disclosed. It relies on developing antibodies to close trichothecene variants that are missing at least one of the hydroxyl groups, and then using these antibodies to test specimens in which the trichothecene has been converted to the variant (usually to the OAC variant). For example, DON and T-2 tetraol can be assayed for using this invention.

Abstract: In an assay, a conjugate of ligand and sac lysing agent is contacted with analyte and binder to produce bound and unbound portions of conjugate. The unbound portion of the conjugate contacts sacs, containing a detectable marker to release the marker as a measure of analyte. The binder and sacs may be placed on different portions of a solid support to provide a solid phase assay.

Abstract: A specific hybridoma cell line produces monoclonal antibodies which are effective in detecting carcinoembryonic antigens (CEA). The specific hydribome line and monoclonal antibodes are designated as T84.66-A3.1-H11. The monoclonal antibodies are preferably applied to tissues and fluids to detect the degree of binding of such monoclonal antibodies to such carcinoembryonic antigens.

Abstract: A method for determining the presence of a polynucleotide analyte in a sample suspected of containing the analyte is disclosed. The method comprises combining in an assay medium the sample and first and second polynucleotide reagents complementary to the analyte. Each of the first and second reagents hybridize with a different region of the analyte. The first reagent contains means for rendering the first reagent non-covalently polymerizable. The second reagent contains means for rendering the second reagent detectable. The sample and the first and second reagents are combined in the assay medium under conditions for polymerizing the first reagent wherein the second reagent becomes bound to the polymerized first reagent only when the analyte is present in the sample. A determination is then made as to whether the second reagent has become bound to the polymerized first reagent.

Abstract: Methods and devices for separating bound label from unbound label within an assay mixture and for predispensing assay reactants in self-contained assay vessels, as well as a method for detecting the presence and/or amount of an analyte within a fluid sample, and a reusable detection vessel for use therein and with specific binding assays in general are disclosed. Significant to the separation of bound label from unbound label is the use of a cushion comprising generally one primary layer and in some cases one or more secondary layers.

Abstract: An immunoassay process for quantitative analysis of an analyte antigen (or analyte antibody) in an aqueous liquid sample. In the first step, the analyte antigen (or analyte antibody) is allowed to react with an antibody (or antigen) in competition with a known quantity of a labelled antigen (or labelled antibody), the antibody (or antigen) being carried by water-insoluble microparticles in the immobilized state. In the second step, the reaction mixture is spotted on a top porous layer of an integral multi-layered analysis element which includes the top porous layer, an intermediate detection layer and a bottom water-impermeable and transparent support. The top porous layer permeates aqueous solution and has pores sufficiently small to trap all of the microparticles carrying the antibodies (or antigens) bound to the analyte antigen (or analyte antibody) and bound to the labelled antigen (or labelled antibody).

Abstract: In an assay wherein there is produced in an assay sample tracer bound to a soluble binder and unbound tracer, both bound and unbound tracer are separated from the assay sample for separate determination thereof by use of a supported binder for the soluble binder and a supported binder for the tracer. The assay may be effected as a "flow-through assay" by causing the sample to flow through successive chambers containing the supported binders.

Abstract: A method for immunological assay of IgM antibodies against at least one said antigen in a liquid sample containing at least IgM and IgG antibodies, such as a biological fluid, comprising reaction of said liquid sample and of antibodies against IgG, and addition to the so obtained reaction product of the said antigen bound to finely divided particles and of a free said antigen, the amount of IgM being inversely proportional to the amount of unagglutinated finely divided particles.

Abstract: The present invention provides a process for the preparation of an immuno-reactive, porous carrier material by application of a solution of a first reaction component of an immuno-reaction and of a solution of a second component of an immuno-reaction coprecipitating therewith, incubation of the carrier material impregnated with the solutions for the immuno-precipitation, optional washing and subsequent drying of the impregnated carrier material, wherein a solution of both components of the immuno-reaction is prepared, which solution contains an inhibitor for the immuno-precipitation, the carrier material is impregnated with this solution and then the immuno-precipitation is initiated by removal of the inhibitor or by removal of the inhibiting action.

Abstract: A method and composition for use in preparing red blood cells for use as indicator cells to detect the presence of antigens and antibodies in biological fluids is the subject of the present invention. Freshly collected human or animal erythrocytes are washed six times with isotonic saline, and suspended to a hematocrit of four percent (volume/volume) with isotonic saline. An incomplete antibody, specific for the antigen or antibody to be detected, in a quantity less than that which would cause irreversible agglutination, is added to the erythrocyte solution. The mixture is incubated, washed, and suspended to a hematocrit of 3-5 (v/v) with isotonic saline. A second antibody, immuno specific for the first antibody and in an amount sufficiently great to immunologically bind substantially all of the antibody carrying red blood cells but not so great as to cause irreversible agglutination, is attached to the erythrocytes. This mixture is incubated, washed, and suspended to a hematocrit of 0.

Abstract: Novel compositions and methods are provided for detecting lupus nephritis in patients suffering from SLE. Competitive or non-competitive assays may be employed, where monoconal antibodies may be used to compete with antiserum for a nucleic acid ligand or may be used for the detection of antiidiotype antibodies as diagnostic of the presence or absence of lupus nephritis.

Abstract: Binding assay methods involving determining the presence of analytes in samples through enzymatic formation of detectable substances in amounts related to the amount of analyte present in the sample and monitoring for the presence of the substances in distinct phases. Methods according to the invention include use of labelled materials which associate with the analyte to be determined or compete with the analyte for association with an added binder. The labelled materials employed include label portions which enzymatically form substances from substrates provided in or existing as a first phase, or, upon enzymatic treatment in a first phase, disassociate into substances capable of existing in or as a second distinct phase. Formation of the detectable substances is monitored by determining the transfer of the substance to a second distinct phase in contact with the first phase or by determining formation of a second distinct phase.

Abstract: Alzheimer's Disease (AD) sufferers may be diagnosed by examination of T-cells from their peripheral blood. AD is marked by increased appearance of alpha-1 acid glycoprotein (AGP) on the surface of such cells. Among AD subjects normal for AGP, a subpopoulation exists which show elevated AMLR response as compared to age-matched controls. These tests may be used to diagnose AD or to screen possible therapeutic agents.

Abstract: Murine monoclonal antibodies specific to the tumor marker galactosyltransferase II (GT-II) which have no measurable cross-reactivity with galactosyltransferase I (GT-I) are described. These antibodies are used in assays to determine levels of GT-II in serum or other body fluid samples, which levels, depending on their magnitude, are indicators of cancer.

Abstract: A method for performing a diagnostic immunoassay by solid phase separation for digoxin. To a reaction mixture of a test sample and labeled anti-digoxin antibody, which forms a complex of any digoxin present in the test sample, is added a solid phase material having an immobilized ouabain triacetate derivative compound capable of binding any excess labeled antibody. The solid phase material is chosen to rapidly settle whereby a solid and liquid phase is formed. The liquid phase can then be extracted to measure the amount of digoxin-labeled antibody present therein. Ouabain triacetate derivative compounds possess sufficient affinity for anti-digoxin antibodies, and are therefore useful in a solid phase separation based digoxin immunoassay for settling out such antibodies without contributing to undesired background interference.

Abstract: Disclosed are 1,4-dihydropyridine immunogen conjugates of immunogenic carrier materials coupled to a 1,4-dihydropyridine derivative. Said conjugates are useful for the preparation of antibodies thereto which antibodies may be used in immunoassays for 1,4-dihydropyridine compounds. An affinity column useful for the purification of said antibodies is also disclosed.

Abstract: An improvement in an assay method which relies on the detection of a labeled, solubilized specific binding complex is achieved by linking the label to a component of the complex through a bond cleavable under conditions compatible with the assay and with the complex in solution.

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: Method of determining changes occurring in articular cartilage. The method involves (a) quantifying proteoglycan monomer and/or antigenic fragments thereof in a synovial fluid sample and (b) correlating the values thus obtained with progressive destructions in the articular cartilage appertaining to that sample fluid.

Abstract: A new and useful fluorescent chromophore has been isolated and identified which has been observed in proteins exposed to glucose over time, and whose fluorescent properties closely resemble those of the polypeptide after it undergoes advanced glycosylation. The chromophore has been structurally identified and named 2-furoyl-4(5)-(2-furanyl)-1H-imidazole, and is believed to be one of the end products of extended nonenzymatic polypeptide glycosylation, which results in the state known as nonenzymatic browning (NEB). The measurement of this chromophore makes possible both qualitative and quantitative assessment of the degree of aging. Diagnostic and test kits are also disclosed.

Abstract: The use of immunologically non-specific peptide linked amino acids containing compounds that are capable of immobilizing circulating immune complexes for the purpose of detection or removal from serum or blood, such compounds including oligopeptides, modified oligopeptides, polypeptides, modified polypeptides, proteins, modified proteins, and in particular glycosylated polypeptides and proteins.

Abstract: Methods and compounds are disclosed for determining the presence, amount of, or association between substances of interest in samples suspected of containing same. The methods are based on the polymerization-induced separation of specifically-bound, reporter-labeled recognition reactants from free, reporter-labeled recognition reactants. The methods described are applicable to any substance for which suitable recognition reactants exist or can be made and are not limited by considerations such as chemical composition or molecular size.

Abstract: The present invention provides a process for the preparation of digitalis antibodies in which appropriate mammals are immunized with a digoxin bound to protein, the animal serum is obtained, the immune globulin-containing protein fraction is separated in known manner, the immunologically-active globulins are adsorbed on an immunologically-active column and separated from the other proteins, the antibodies are again eluted from the column and the Fab fractions are split with papain and purified, wherein, as immunologically-active adsorbent, there is used an inorganic matrix of large surface area to which digitoxin aldehyde is bound via a spacer which cannot be split with papain and the splitting off of the Fab fraction from the antibodies is carried out on the matrix.The present invention also provides digitalis antibodies obtained by this process, which antibodies can be used for the therapy of digitalis intoxications and for preparing immunological reagents for the determination of digitalis glycosides.

Abstract: A process for producing an antibody having a specificity to estriol-3-sulfate, which comprises administering a 6-substituted-estriol-3-sulfate protein conjugate of the formula: ##STR1## wherein A is .dbd.N--O-- or --O--CO--, n is an integer of 1 to 4 and --NH--P is the residue of a protein excluding a hydrogen atom in the amino form therefrom parenterally to a living body of a vertebrate animal so as to produce the antibody in the living body and collecting a body fluid comprising the antibody from the living body.

Abstract: A method is disclosed for the determination of human serum factors which are specific for human carcinomata antigens, in which method the serum of a patient with a carcinoma is incubated with CEA that has been radioactively labeled and anti-goat immunoglobulin serum in toto or depleted of the cross-reaction capacity for human IgA, IgG and IgM is added, then the whole mass is incubated and centrifugated, the supernatant is decanted and the precipitate is counted with a gamma counter. These specific serum factors are employed as a marker of primitive tumoral forms.

Abstract: A method for assaying complement fragment C.sub.3 a, C.sub.4 a or C.sub.5 a or the des-Arg derivative thereof in a biological sample which comprises combining equal volumes of the biological sample and a solution of 0.8 to 1.6% of an acridine derivative selected from the group consisting of acrinol, acriflavine, acriflavine hydrochloride, and aminacrine, incubating the mixture for about one minute to 2 hour at about 25.degree. C., recovering the supernatant from the resultant precipitate, incubating the supernatant with a known amount of a labeled complement fragment selected from C.sub.3 a, C.sub.4 a or C.sub.

Abstract: Fluorometric immunological assay method in which an antigen is attached onto magnetic particles, which are, for the time of measurement, pulled against the wall of the measurement vessel by means of the magnetic field. The magnetic particles adhering to the vessel walls are irradiated and the resultant emitted fluorescence is measured.

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,This invention was supported in part by Grants No. CA-15126 and CA-15437 from the National Cancer Institute, U.S. Public Health Service.