Abstract: An improved method for performing immunoassays whereby specific binding proteins for vitamin B12, folate and other target analytes are utilized with antibodies with different specificities for the binding proteins. Antibodies bridge the specific binding protein directly or indirectly to a capturable material.

Abstract: Disclosed herein are methods for screening for primary cardiomyopathy. The methods are preferably immunological methods in which the level of binding of a monoclonal or polyclonal antibody to a 50 kD glycoprotein component of a mammalian muscle tissue is determined. This level of binding is compared to the level of binding observed when non-dystrophic tissue is treated in an otherwise identical manner. A substantial reduction in the level of binding to the 50 kD glycoprotein in the experimental mammalian muscle tissue has been determined to be a screen for primary cardiomyopathy.

Abstract: The present invention provides a process for the determination of low density lipoproteins (LDL) in body fluids, wherein high density lipoprotein (HDL) antibodies are added to a sample to be investigated, insolubles formed are separated off and the LDL or one of its components is determined in the supernatant.The present invention also provides a reagent for the determination of the LDL fraction in body fluids, wherein it contains HDL antibodies.

Abstract: The present invention provides an improved buffer solution for conducting an immunological method for measuring apoprotein of human tissue factor, and a kit therefor.

Abstract: A method of immunoanalysis combines immobilized immunochemistry with the technique of flow injection analysis, and employs microscopic spherical structures called liposomes, or lipid vesicles, as carriers of detectable reagents. Liposomes are modified on their surface with analytical reagents, and carry in their internal volume a very large number of fluorescent or electroactive molecules. Aspects of this embodiment of the invention include the chemistry for covalent immobilization of antibody fragments in a specified orientation, the use of liposomes in a flow injection analysis system, and the combination of automated sampling and analysis with reusable immunoreactants. Another aspect of the invention involves the non-covalent binding of liposomes to a receptor for use in a homogeneous assay. In another aspect of the invention the intensity of scattered light is quantitated as a measure of liposome aggregation in response to a concentration-dependent immunospecific reaction.

Abstract: Disclosed are a method and device for performing sequential analytical reactions involving a first dry reagent and a second dry reagent comprised of two or more components having different rates of solubilization. The invention enables one to fully solubilize the components of the second reagent before they are brought into contact with each other to thereby avoid interference with the reaction kinetics which result when one or both of the components are not fully dissolved prior to their being brought into contact. The invention is especially useful in conjunction with immunoassay formats involving latex bound antibodies and polymeric agglutinators.

Abstract: The present invention first provides monoclonal antibodies recognizing membrane phospholipase A.sub.2, namely, monoclonal antibodies PL-49, PL-71, PL-76, and PL-78, hybridomas producing them, methods for producing them, and immunoassays of membrane phospholipase A.sub.2 using them.The immunoassay of PLA.sub.2 M is useful for the diagnosis of articular rheumatism, cancers, and a wide variety of inflammatory states.

Abstract: Pancreatic disease can be diagnosed by assaying a patient's body fluid such as serum or urine, for pancreatic activation peptides (PAP) released from zymogens by proteolytic activation. Particularly useful are peptides having C-terminal D.sub.4 K sequences. The method uses polyclonal or monoclonal antibodies generated and selected for C-terminal specificity on PAP so that the tests only report free PAP not parent zymogen. Also described are peptides and antibodies labelled with revealing agents and/or immobilised on solid supports and their use in diagnostic assays and kits. In pancreatic disease the tests distinguish necrotising from oedematous acute pancreatitis and permit severity prediction and monitoring as well as diagnosing chronic pancreatitis in exacerbation.

Abstract: A method for determining whether antiphospholipid antibodies are present in a sample is disclosed. The method comprises contacting the sample with a negatively charged phospholipid and with .beta.-2-glycoprotein-I or a homolog or analog thereof and determining whether any antiphospholipid antibodies have bound to the contacted phospholipid and .beta.-2-glycoprotein-I, wherein detection of binding of antiphospholipid antibodies to the phospholipid and .beta.-2-glycoprotein-I, is indicative that antiphospholipid antibodies are present in the sample.

Abstract: Dopamine releasing protein (DARP) is a protein which stimulates release of dopamine from dopaminergic neurons, and is effective at extremely low concentrations. DARP is useful for stimulating vertebrate nervous systems, and for treatment of Parkinson's disease. Antibodies to DARP are useful for inhibiting dopamine release, and for quantifying the concentration of DARP in samples.

Abstract: Disclosed are immunofluorescence staining methods which increase the likelihood that antibodies expressed by a single B cell selected and sorted by fluorescence activated cell sorting are specific for the antigen of interest, and which also allow selection of B cells expressing antibodies of high affinity for the antigen of interest. The selection for B cells expressing antibodies to specific antigens is increased by labeling B cells with at least two antigen probes, where each antigen probe includes the antigen of interest and is labeled with a different fluorochrome. The positive selection is preferably combined with a negative selection step, in which autofluorescent cells and sticky cells are excluded out. The specificity of sorting of the desired B cells can be further enhanced by staining those antigen-specific B cells which produce the immunoglobulin isotype (typically IgG), with targeting molecules reactive with a B cell marker, such as .gamma.

Abstract: A competitive protein binding assay for rapamycin and biologically-active metabolites, derivatives and analogues thereof in blood and other biological fluids is disclosed, wherein the binding reagent is a specific rapamycin binding protein either substantially purified from the soluble cytoplasm of target cells of rapamycin action, particularly normal or transformed lymphocytes, freshly collected or in established cell lines, or synthesized by recombinant DNA techniques. Solution phase and solid state assay systems are disclosed.

Abstract: A method is provided for detecting T-cell dysfunctions. The method includes detecting an alteration in the level of a protein regulating synthesis of the hexasaccharide NeuNAc.alpha.2.fwdarw.3Gal.beta.1.fwdarw.3(NeuNAc.alpha.2.fwdarw.3Gal.beta .1.fwdarw.4GlcNAc.beta.1.fwdarw.6)GalNAc on leukosialin of T-cells from a subject suspected of having a T-cell dysfunction compared to resting T-cells from a normal individual. The protein regulating synthesis of the hexasaccharide can be core 2 GlcNAc transferase and can be detected by either a change in its amount or activity. Also provided is a method of detecting T-cell dysfunctions which includes detecting an alteration in the level of leukosialin having the hexasaccharide NeuNAc.alpha.2.fwdarw.3Gal.beta.1.fwdarw.3(NeuNAc.alpha.2.fwdarw.3Gal.beta .1.fwdarw.4GlcNAc.beta.1.fwdarw.6)GlcNAc on T-cells from a subject suspected of having a T-cell dysfunction compared to resting T-cells from a normal individual. Kits for detecting T-cell dysfunction are provided as well.

Abstract: This invention relates to methods that have been found useful in reducing non-specific binding in immunochemical assays, via methods that can be implemented much faster than those used by Ishikawa. The techniques include the use of a lipophilic bridge, such as a liposome, or the elution of the antigen-antibody complex from the solid phase by the use of an anti-idiotypic antibody or an antibody, or the use of a heterologous reversible bridge.

Abstract: An enhanced chemiluminescent assay, in which a dihydrophthalazinedione such as luminol, a peroxidase such as HRP and an oxidant such as H.sub.2 O.sub.2 are co-reacted in the presence of an enhancer such as p-iodophenol, is modified. The enhancer is generated by enzyme-catalysed reaction of a pro-enhancer, e.g. p-iodophenol phosphate is cleaved by alkaline phosphatase, enabling this enzyme to be assayed instead of peroxidase. Alternatively, the enhancer is added, an anti-enhancer such as p-nitrophenol is generated by enzymatic reaction of a pro-anti-enhancer such as p-nitrophenol phosphate and the reduction in luminescent emission is measured.

Abstract: This invention uses the evanescent wave detection of particles to distinguish bound from free in an analyte-binding assay. Illumination below the critical angle is employed, and a beveled window is used to eliminate bulk fluorescence from the emitted evanescent wave liquid. The sample is held in a non-rigid film cuvette.

Abstract: The invention concerns a method for detecting an analyte in a blood sample or a derived fraction thereof. In this method one uses the binding affinity between the analyte and its specific binding agent. Moreover, a reaction component containing a peroxidase label is added to the sample and specific binding agent to form a reaction mixture. The incubation of the reaction mixture, containing the analyte and its specific binding agent, with the peroxidase-labeled reaction component has to be carried out in the presence of a quaternary ammonium salt or ionic surfactant.

Abstract: The present disclosure relates to a new lymphokine molecule, referred to as Monocyte Cytotoxicity Inducing Factor (MCF), and its use as in cancer and other types of therapy. The disclosure also relates to antibodies that immunologically recognize MCF, as well as to the use of such antibodies in detecting the presence of MCF and biologic samples. The disclosure further relates to the development of novel Sezary cell hybridomas which secrete MCF and thereby provide a ready source for MCF isolation and purification.Sezary's Syndrome is a leukemic proliferation of OKT4+ lymphocytes. Sezary cells were isolated by differential centrifugation and fused to CEM.8aza.sup.r.C, an HGPRTase lacking clone of CEM. The hybrid cells were studied for their ability to produce soluble mediators of human monocyte cytotoxicity. The product of a single clone, FtF3, which bore the surface phenotype of Sezary cells, was characterized.

Abstract: A method and apparatus are provided for selecting and analyzing a subpopulation of cells or cell objects for a certain parameter such as DNA using image analysis means. The cells are first stained with an alkaline phosphatase technique including a monoclonal antibody specific to a protein in at least one of the cell's cytoplasm or on a cell membrane, thereby marking any cells including the protein as to type. A second staining of the DNA in the nucleus is accomplished by a Feulgen technique that destroys the cell cytoplasm. After the staining and marking, the cells may then be gated using the image analysis means on the visual parameter such as colored DNA or colored antigen into a subpopulation that is to be measured. The selected cells may then be examined by digital image processing and measured for a parameter such as a true actual measurement of DNA in picograms. A quantitation of the measured parameter may be generated and provided.

Abstract: Methods are provided for detecting receptivity of mammalian endometrium to embryo implantation comprising obtaining a sample of the endometrium, contacting the endometrium with a monoclonal antibody for .beta..sub.3 integrin and detecting .beta..sub.3 integrin in the endometrium. The invention also provides for methods of diagnosing infertility in a mammal and methods of detecting the window of embryo implantation in endometrium. Methods of in vitro fertilization, methods of preventing embryo implantation and a method of monitoring endometrial maturation are also within the scope of the present invention. The present invention is also directed to contraceptives. Diagnostic kits useful in the practice of the methods of the invention are also provided.