Abstract: The invention describes reactive dyes having an alkyl spacer attached via a sulfonamide bond to a sulforhodamine 101 fluorophore, and a variety of useful conjugates prepared therefrom. The increased length of the covalent linkage due to the alkyl spacer results in dye-conjugates having a number of surprisingly advantageous properties relative to previous sulforhodamine 101-labeled conjugates, including enhanced solubility and increased fluorescence. The reactive dyes of the present invention are more stable than the known compound sulforhodamine 101 sulfonyl chloride. Novel reactive dyes are described for selective modification of groups other than amines, including thiols and photoreactive derivatives.

Abstract: A solid phase method of detection or assay of the presence or amount in a serum or plasma sample of a target antibody specific to a cell surface antigen. The sample is contacted with an immobilised preparation of cells bearing the antigen and antibody bound thereto is detected or assayed by means of an indicator comprising a binding partner for the antibody bound to labelled latex particles.

Abstract: A method and apparatus for detecting rare cells in a biological sample is disclosed. A color image of the sample is generated and the color image is decomposed into its color components. A first mask is then generated based upon a first color characteristic of the rare cells. At least one color component of the color image is filtered using the mask to produce at least a first composite image which contains features having the first color characteristic. At least a second mask is then generated based upon other color characteristics of the rare cells and at least the first composite image is filtered using at least the second mask to create at least a second composite image. The color and/or shape of features in the at least second composite image are analyzed and a list of locations of probable rare cells is generated from the color and/or shape analysis.

Abstract: The present invention provides a method of linking a target particle to an insoluble support, wherein said particle is bound support by means of a specific binding partner, characterized in that the linkage between said binding partner and said support comprises hydroxyboryl/cis-diol bond. The invention has particular utility in the immobilization and isolation of cells.

Abstract: The invention is directed to methods and kits for detecting and measuring the presence or absence of perinuclear anti-neutrophil cytoplasmic autoantibody of ulcerative colitis or primary sclerosing cholangitis. The methods and kits of the present invention provide safe and reliable means for diagnosing ulcerative colitis and primary sclerosing cholangitis. The antigens reactive with perinuclear anti-neutrophil cytoplasmic autoantibody of ulcerative colitis and primary sclerosing cholangitis are also provided.

Abstract: Methods and compositions for detecting schizophrenia based on modification of the dopamine D.sub.4 receptor by addition of an adenosyl group to methionine #313 via methionine adenosyltransferase are described herein. Individuals with schizophrenia have a deficiency in methionine adenosyltransferase activity and a lower amount of modified dopamine D.sub.4 receptor than normal individuals. Methods for screening therapeutic processes, agents and drugs for use in treatment of schizophrenia are also described.

Abstract: Compounds and libraries are labeled with a galactosyl epitope amd then screened in accordance with an assay involving cells having a characteristic of interest. Conveniently, the screening may embody target cells, where the compounds are brought in contact with the cells. Each of the compounds carries with it the information of its identity or method of synthesis. After washing away non-specifically bound compounds, blood may be applied to the cells, whereby antibody binding to the galactosyl epitope initiates the complement cascade. Plaques are identified and the compound associated with the plaque identified. The formation of the plaque demonstates that the compound has specific affinity for the target cell, binding of the compound to the cell does not interfere with binding of the antibody, and that the complex is capable of cytotoxic activity by means of the complement cascade.

Abstract: A method for detecting presence or absence of a nucleic acid of interest in fetal nucleic acid derived from a sample of peripheral blood obtained from a pregnant woman is described. The method involves obtaining a sample of peripheral blood from a pregnant woman, treating the sample of peripheral blood such that fetal nucleic acid present in fetal granulocytes is made available for detection and detecting presence or absence of a nucleic acid of interest in the available fetal nucleic acid. The proportion of fetal granulocytes present in the sample of peripheral blood can be increased relative to the sample of peripheral blood forming a sample enriched in fetal granulocytes prior to the detection step.

Abstract: Methods and compositions are provided for use of modified or intact phycobilisomes as extremely potent labels in sensitive monitoring kits (e.g., for blood contamination), specific binding assays (e.g., visual, photometric and fluorometric immunoassays) and optoelectronic devices (e.g., biosensors, photoelectric transducers).

Abstract: Methods and compositions for detecting schizophrenia based on modification of the dopamine D.sub.4 receptor by addition of an adenosyl group to methionine #313 via methionine adenosyltransferase are described herein. Individuals with schizophrenia have a deficiency in methionine adenosyltransferase activity and a lower amount of modified dopamine D.sub.4 receptor than normal individuals. Methods for screening therapeutic processes, agents and drugs for use in treatment of schizophrenia are also described.

Abstract: An immunochemical method is provided for the detection and determination of an analyte. The zeta potential of a latex-particle loaded with an immunologically active substance is measured before and after bringing the loaded latex-particle into contact with an analyte. The difference in zeta potential is correlated with changes of zeta potential for known concentrations of the analyte in order to determine the presence and amount of the analyte.

Abstract: The CNS myelin associated proteins inhibit neurite outgrowth in nerve cells and neuroblastoma cells, and can also inhibit fibroblast spreading. Such inhibitory proteins include a 35,000 dalton and a 250,000 dalton molecular weight protein and analogs, derivatives, and fragments thereof. The CNS myelin associated inhibitory proteins may be used in the treatment of malignant tumors. The present invention is also directed to antibodies to the CNS myelin associated proteins; such antibodies can be used in the diagnosis and therapies of nerve damage resulting from trauma, infarction, and degenerative disorders of the central nervous system. In a specific embodiment of the invention, monoclonal antibody IN-1 may be used to promote regeneration of nerve fibers over long distances in spinal cord lesions.

Abstract: A method for detecting a target substance which includes collecting a substance sample; introducing the substance sample into a substance card having at least one reagent responsive to the presence of the target substance and having a light-transmissive chamber; inserting the substance card into a substance detector device having a photosensor and adapted to receive the substance card; mixing the substance sample with the reagents for a preselected mixing period, thus producing a measurand having a target substance reaction; streaming the measurand through the light-transmissive chamber and illuminating the measurand with a quantity of light for a selectable analysis period, an optical characteristic of the measurand can be measured with the photosensor; and selecting the target substance reaction as a positive reaction or a negative reaction, responsive to the optical characteristic. If the target substance reaction is a negative reaction, the device operator is signalled thereof.

Abstract: Derivatives of 2-nitro-imidazole, such as 1-(2-hydroxy-3-piperidinopropyl)-2-nitro-imidazole, are useful as hypoxic cell markers for immunochemical detection of hypoxia in tumor tissue.

Abstract: Methods for detecting and isolating acrosome-reacted sperm and complement receptor-bearing oocytes using the complement component C3, fragments, or variants thereof, antibodies to a complement receptor, or antibodies to C3, are disclosed. These methods have application in the assessment of fertility, in the preparation of sperm or oocytes for in vitro fertilization or for gamete intrafallopian tube transfer, in promoting or inhibiting fertilization in vitro and in vivo, and in diagnosing and treating infertility.

Abstract: A coupling agent which is an activated ester such as N-maleimido-6-aminocaproyl-HNSA (mal-sac-HNSA) is formed by reacting 4-hydroxyl-3-nitrobenzene sulfonic acid sodium salt (HNSA) with a carboxylic acid moiety of a compound such as N-maleimido-6-aminocaproic acid. The coupling agent is reacted with an amino group of an amine-containing biological material such as a protein at a pH of about 5.5 to 10.0 and HNSA is released. The released HNSA is spectroscopically measured at a wavelength of from about 350 nm to about 500 nm to precisely monitor and control conjugating of the coupling agent to the biological material. The resulting product is coupled to a sulfhydryl group or other group of another material to provide cross-linking between the two. This enables joining the biological material to one another, to a support matrix, to a label, to a hapten, and to other materials.

Abstract: A fluorescent labelling composition comprises a linear polysaccharide backbone molecule having a plurality of target-binding molecules, such as antibodies or nucleic acids, attached at spaced-apart intervals thereon. Each of the target-binding molecules, in turn, includes a multiplicity of fluorescent dye molecules bound thereto. In this way, fluorescent signal introduced to a single target-site on a solid phase surface may be increased without loss of binding activity.

Abstract: This invention provides a method for preserving cells which comprises the steps of (a) suspending cells in a physiologically-acceptable, isotonic medium; and (b) fixing the cells so suspended at a temperature of less than about 10.degree. C. under sufficiently hypertonic conditions so as to disperse the cells in a single, unagglutinated state, thereby preserving cells. This invention also provides a method for detecting cells separated from a sample which have been preserved according to the aforementioned method. This invention also provides a method for visualizing cells. Also provided is a method for detecting a metabolic process in cells present in a sample. This invention also provides a method for detecting the presence of rare cells in a sample which specifically possess on their surfaces a moiety recognized by a known ligand comprising preserving cells separated from the sample according to the aforementioned method for preserving cells.

Abstract: Disclosed are materials and methods for detecting biomolecules in samples employing transponders having memory elements associated with particle(s) used as a solid phase in art assay, and information pertinent to the assay is encoded on the transponder memory elements. A dedicated read/write device is used remotely to encode or remotely to read the information encoded on the transponder memory elements. The invention can be used in direct or competitive ELISA-type assays, or in multiplex assays for the simultaneous assay of several analytes.

Abstract: The present invention relates to a method of determining the presence of an antigen, particularly the HLA-B.sub.27 antigen, at the surface of cells, particularly lymphocytes, by combining the sensitivity of a cytotoxicity test, particularly a lymphotoxicity test, combined to a fluororescence detection, particularly, by using propidium iodide. The claimed method does not involve any purification of lymphocytes from a blood sample. The erythrocytes contained in the test blood sample are lysed after the immune reaction and complement action is completed. Moreover, the formation of the immune complexes, the complement fixation and reaction and the staining of the cells may take place at the same time in the test tube. This method is therefore very simple and very fast to execute. The facility of reading the positive cells is also an advantage to the claimed method over and above the methods using dyes like eosin or trypan blue, because of the direct visualization of colored cells as positive cells.

Abstract: HIV-1 peptides having at least one point mutation between position 593 and 611 of the HIV-1 gp160 amino acid sequence. The point mutation either is at position 604 or 610, or both positions. Immunoassays which utilize these peptides are provided, as well as, diagnostic test kits which contain these peptides.

Abstract: The invention is an assay, including a series of peptides, which allow screening for inhibitors of C5a binding targeted to the subsite on the C5a receptor occupied by the C-terminus of C5a. These peptides allow compound testing efforts to be targeted to this same subsite so that C5a agonists and antagonists can be identified. These peptides have much greater affinity (Ki<10 nM) than does the natural C-terminus of C5a (Ki=300 .mu.M) and have been labeled to allow for detection of molecules which inhibit binding of these peptides at this receptor subsite. The invention is useful to develop agonists, partial agonists, and antagonists of C5a, and the invention includes compounds identified according to the method of this invention and methods of their use.

Abstract: A process for preparing a purified syphilis antigen which comprises adsorbing an extract originated from Treponemda pallidum on a hydroxyapatite gel, followed by elution, while an aqueous medium is used.

Abstract: Drug-dependent antibodies that bind to granulocytes, erythrocytes, platelets or membrane proteins derived from these cells, in the presence of a drug, but not in its absence, can be detected using a sensitive assay. Detection of the drug-dependent antibodies permits diagnosis of cytopenia mediated by the drug.

Abstract: A method for detecting a component of an aqueous liquid comprising adding to the liquid a luminescent dye selected from the group consisting of cyanine, merocyanine and styryl dyes containing at least one sulfonic acid or sulfonate group attached to an aromatic nucleus and reacting the dye with the component. The labeled component is then detected by an optical detection method.

Abstract: A method of identifying compounds that effect integrin activation in intact cells is provided.

Abstract: A flow cytometric method for determining procoagulant platelet-derived microparticles in whole blood is described.

Abstract: A novel monoclonal antibody of IgG1/.kappa. subclass, characterized in that it recognizes antigens specifically present in mouse inbred strains consisting of DBA/1, CE/J, SM/J, IQI, PL/J, SWM/Ms, RIIIS/J, RFM/MsNrs and C3H and its congenic mouse strains thereof, wherein one of said antigens has a molecular weight of 66,000 and the other has a molecular weight of 68,000 and said antigens are present in the mitochondria of C3H mouse strain.The present invention also provides a hybridoma cell line which produces the above monoclonal antibody, a method for identifying the origin of a cell derived from a mouse, and a reagent for identifying the origin of a cell derived from a mouse.

Abstract: A method for the detection of nucleic acid-containing moieties is described which combines affinity capture of the moiety with detection and identification of the moiety's nucleic acid.

Abstract: A suspension of inert particles is prepared in an aqueous solution, to which an antibody or an antigen and a carrier-bound antigen or antibody, respectively, are added in any desired order. After centrifuging, the positive, weakly positive, or negative reaction can easily be recognized on the basis of a simple pattern.

Abstract: Desired cells are positively separated from a mixture of cells using multiple stages of affinity surfaces. Bound cells from each surface are removed and subjected to a further surface for further enrichment.

Abstract: A vessel for conducting blood cell agglutination assays is disclosed. A barrier retains reactants in an upper chamber during incubation, then, in response to a force, permits reagents to enter a lower chamber containing a matrix for separating agglutination.

Abstract: Methods of screening for endometriosis comprise obtaining a sample of endometrium selected from menstrual cycle day 20 to 24, identifying the endometrial sample as nulliparous, contacting the endometrium with a monoclonal antibody for .beta..sub.3 integrin, assaying for .beta..sub.3 integrin and correlating the absence of .beta..sub.3 integrin with endometriosis, wherein the endometrium is identified as mild/minimum endometriosis. A method of using monoclonal antibodies to screen for endometriosis is also within the scope of the invention. Methods 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 and detecting .beta..sub.3 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.

Abstract: Methods for detecting and isolating acrosome-reacted sperm and complement receptor-bearing oocytes using the complement component C3, fragments, or variants thereof, antibodies to a complement receptor, or antibodies to C3, are disclosed. These methods have application in the assessment of fertility, in the preparation of sperm or oocytes for in vitro fertilization or for gamete intrafallopian tube transfer, in promoting or inhibiting fertilization in vitro and in vivo, and in diagnosing and treating infertility.

Abstract: An article for performing solid-phase immunological assays and methods for performing assays utilizing such article. A solid-phase support suitable for immunological assays is stained with an organic dye that has a net-positive charge and a hydrophobic aromatic ring structure. An immunologically reactive component having a net-negative charge, such as a whole cell, is securely immobolized by the dye to the support through noncovalent interactions. A sample of biological fluid may be added to the article to which the immunologically reactive component has been immobilized to effect binding of antigens or antibodies in the sample which are specific to the immobilized component. The presence of the antigen-antibody complex may then be determined by any known means.

Abstract: Methods and cards for detecting an antigen or antibody in a fluid sample are disclosed. The fluid sample is added to a microreaction vessel containing a slurry suspension of inert particles and a binding partner to the antigen or antibody to be determined. The fluid sample is added to the micro reaction vessel, with one of the antigen and antibody binding partners being carrier bound. The vessel contents are centrifuged, to cause the binding partners to contact each other to form an optically detectable binding complex. The location of the optically detectable complex is observed to determine the presence of the antibody.

Abstract: Advantage is taken of the ability of rotaviral VP6 protein to home to macrophage and monocytes to provide label to these cells in either an in vitro or in vivo environment. Further, the ability to couple label to the VP6 protein and to couple VP6 to a targeting agent provides a mechanism for conducting label (or an effector moiety) to any desired target.

Abstract: Apparatus for performing immunoassays which is essentially self-contained, requiring only the introduction of a sample and, at appropriate times, washing solution. The apparatus (10) includes: a fluid container (12) having a central platform area with a reaction area (30) which can contain a reactive agent; a sample receiving chamber (22) having a sample conduit (24) located above the porous medium; at least one openable reagent container (46); a conduit (28) for directing reagents onto the porous medium; an opening member (50) attached to the upper unit and positioned to contact and open reagent containers sequentially, by incremental relative rotation of the upper and base units; and a window (34) for viewing the reaction area. The apparatus can also include a sampler member (58) in the nature of a tampon for assays involving samples taken from body cavities.

Abstract: The present invention relates generally to test articles and assays for the detection of analytes in biological fluid samples. More particularly, the present invention relates to test articles an assays which employ dyed microorganisms as visual labels to detect suspected analytes.

Abstract: In an erythrocyte agglutination assay, the agglutination reagent comprises at least one erythrocyte binding molecule coupled to at least one specific analyte binding molecule wherein the erythrocyte binding molecule does not cause agglutination when incubated with erythrocytes in the absence of analyte (in the case of a direct assay) or analyte binding reagent (in the case of an indirect assay). Preferably, the erythrocytes are endogenous to the blood sample to be tested, that is, a whole blood sample is assayed. Mixtures of conjugates and conjugates of analyte analogues with erythrocyte binding molecules may also be used as agglutination reagents. The reagents and their use in direct or indirect assays is disclosed.

Abstract: A disposable diagnostic unit is provided which employs a housing which provides for a sample port, and a channel which feeds the sample to an incubation area by means of capillary action. The incubation area is underneath an optically-clear window and comprises a lipid membrane which has optical properties, particularly fluorescent properties and usually a reagent. A reservoir at the end of the channel downstream from the incubation area receives the sample and waste washes, while on one side of the platform area is a reagent reservoir and on the other side a side waste reservoir, so that one can move the reagent from the reagent reservoir through the platform area into the waste reservoir. Various reagents may be contained within the unit and the necessary liquids added automatically by appropriate instrumentation, so as to have the assay carried out automatically, without technician involvement, providing an accurate and sensitive determination.

Abstract: Compounds and methods are disclosed for reversibly aggregating particles suspended in a liquid medium. The method comprises combining the liquid medium containing the particles with a polyionic polymer capable of aggregating the particles under conditions suitable for such aggregation. Thereafter, the particles are contacted with a chemical reagent capable of cleaving the polyionic polymer under conditions sufficient to reverse the aggregation. Optionally, magnetic particles are added to the liquid medium in the present method under conditions for non-specific binding and the medium including the aggregates is subjected to a magnetic field gradient to separate the aggregates from the medium. The compounds of the present invention are polyions. The aggregation of the particles is reversible upon contact with chemical agents which cleave at least some of the bonds within the polyionic polymer.

Abstract: By allowing the mast cell enriched population of human conjunctival tissue cells to incubate for a minimum of about forty (40) hours post enzymatic digestion, the treatment window between spontaneous histamine release and anti-human IgE stimulated histamine release is increased. Culturing the human conjunctival tissue mast cells decreases the spontaneous release of histamine and increases the anti-IgE stimulated histamine release, greater than ten fold over spontaneous, at time points over forty (40) hours. This treatment window is sufficient to detect a compound's stabilizing or anti-allergic activity.

Abstract: A method of detecting target antibodies or antigens by reaction with specific binding partners thereto is disclosed. One of the target or the binding partner is bound to a carrier, and the other is unbound. The complex between the target and the binding partner, with one being carrier-bound, forms an optically detectable binding complex. A microreaction vessel having an upper portion, a transition portion and a lower portion is utilized, wherein the upper portion has a greater diameter or width than the lower portion, and the transition portion is situated between the upper portion and the lower portion, and is funnel shaped. The microreaction vessel contains a slurry or suspension of inert particles, and unbound target or binding partner thereto. A solution of the carrier bound target or binding partner thereto is added to the vessel, which is then centrifuged to produce an optical determination of the target.

Abstract: A chemical moiety is disclosed which comprises a chemical, biochemical, or biological substance attached to one or more electrochemiluminescent organometallic compounds. In a preferred embodiment of the invention the substance is attached to one or more ruthenium-containing or osmium-containing luminescent organometallic compounds. Methods are disclosed for detecting low concentrations of the chemical moiety using chemiluminescent, electrochemiluminescent, and photoluminescent means. Compounds are disclosed which are useful for labeling substances of interest with ruthenium-containing and osmium-containing labels or other electrochemiluminescent labels. These labeled substances are useful in methods provided for detecting and quantifying analytes of interest in binding assays and competitive binding assays. The labeled substances are of particular use in homogeneous binding assays.

Abstract: The present invention relates to an improved method for the production of antibodies to tumor-associated gangliosides using ganglioside lactones. The resulting antibodies are useful in the detection and treatment of tumors containing gangliosides. The present invention also relates to methods of treatment of tumors by active immunization using ganglioside lactones.

Abstract: By cloning the gene for particular keratin proteins, it has been possible to determine amino acid sequences of specific keratins. This has made possible selection of sequences which are unique to a given keratin. The production of antibodies that respond to selected sequences provides means of selectively identifying specific keratins. These diagnostic tools provide means of identifying cell source of malignancies.

Abstract: Novel hybridomas, human monoclonal antibodies and their uses are provided. The antibodies distinguish a human neoplastic cell from a normal cell of the same tissue type. The monoclonal antibodies find use in therapy and diagnosis, both in vitro and in vivo.

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.

Abstract: A new ligand binding assay is based upon measurements of relaxation rates of the solvent, which are obtained with a magnetic resonance (MR) spectrometer. It is termed a solvent mediated relaxation assay system (SMRAS). SMRAS is based on the observation that the enhancement of proton relaxation rates produced by a magnetic material can be modulated by the binding of various analytes to a magnetic material. Hence the relaxation rate of the solvent can be interpreted to give information on the concentration of an analyte.