Abstract: The present invention relates to iminium ion substituted cyanine dyes having a fluoresence absorbance of between about 500 and 900 nm, a reduced tendency to aggregate and enhanced photostability. The cyanine dyes of the present invention are represented by the formula ##STR1## wherein n is 0, 1, 2 or 3;m is 0, 1, 2 or 3;R.sub.1 and R.sub.2 are taken together to form an aromatic ring or a fused polycyclic aromatic ring;R.sub.3 and R.sub.4 are taken together to form an aromatic ring or a fused polycyclic aromatic ring;R.sub.5 and R.sub.6 are independently selected from the group consisting of (CH.sub.2).sub.p X where p is 1-18 and X is a functional group that reacts with amino, hydroxyl and sulfhydryl nucleophiles;R.sub.7 and R.sub.8 are independently selected from the group consisting of hydrogen, C1-C10 alkyl groups and where R.sub.7 and R.sub.8 are taken together to form a five- or six- membered heterocyclic ring;R.sub.

Abstract: Compounds are evaluated for their binding to naturally occurring receptors, by employing the natural ligand conjugated to an enzyme donor fragment of .beta.-galactosidase for competing with the sample compound for the natural acceptor binding site or in the absence of competition where the sample compound binds to an allosteric site. By adding the enzyme acceptor fragment of the .beta.-galactosidase and substrate, the binding affinity of the sample compound may be evaluated as a measure of agonist or antagonist capability.

Abstract: The invention provides a method for determining platelet function in a mammal comprising providing platelets from the mammal, contacting the platelets in suspension with at least one immobilised ECM protein or an effective fragment or analog thereof while applying to the platelets an effective mechanical stimulus for an effective period of time and determining the platelet activation produced.The invention also provides a method for detecting a bleeding disorder in a human. Further, the invention provides a method for monitoring the efficacy of pharmacological agents affecting platelet function in vivo.

Abstract: A method for detecting a potentially detrimental transformation of cells of an organism. The method comprises determining syndecan content indicator from a sample of biological material from the organism, and comparing the determined syndecan content indicator with a reference syndecan content indicator from a reference biological material of the same type.

Abstract: In a first embodiment, a target analyte in solution is detected by exposing the solution to an electrode that includes a conducting electroactive polymer to which a periodic alternating voltage is coupled. Upon exposure to the analyte, an electrode characteristic is varied, which variation is detected by measuring electrode current as a function of time and as a function of the periodic alternating voltage. The alternating voltage waveform has an oxidizing time period and a reduction time period, which periods and waveform duty cycle may be controlled to enhance electrode sensitivity, selectivity, and to substantially eliminate electrode fouling and data hysteresis. In a second embodiment, a receptor is bound to the electrode, to which is coupled an alternating voltage waveform that permits a mating target substance to reversibly bind to the receptor such that measurement of electrode current provides a measure of such reversible binding.

Abstract: Nucleic acid probes are described for detecting yeasts capable of causing Candida infection, specifically yeasts of the genera Candida, Torulopsis, and Yarrowia. The preferred probes are complementary to ribonucleic acid sequences unique to specific Candida species, or groups of such species, and as such can detect the rRNA, rDNA, or polymerase chain reaction amplification products of these genes. Methods for detecting the etiological agent of human Candida fungemia utilizing these probes are also provided.

Abstract: The present invention provides unique prepared immunogens, site-specific polyclonal antisera and monoclonal antibodies against the DNA-binding domain of estrogen receptor protein, and immunoassay to determine the functional status of estrogen receptors in a cellular sample. Collectively or individually the component parts of the invention provide the ability not only to identify accurately the presence of human estrogen receptor but also the capability of determining whether the estrogen receptor exists in a functional or non-functional state.

Abstract: This invention relates to a method of determining the amount of test analyte in a sample using internal calibration comprising mixing a sample with a predetermined amount of a calibrator analyte foreign to the sample and with a comparable behavior in an assay to that of the test analyte, contacting the mixture with a solid support containing, each in a separate area, a regent for binding the test and calibrator analytes, respectively, contacting the solid support with a mixture of labelled reagents for binding the test and calibrator analytes, respectively, and determining the amount of test analyte in the sample by comparing the levels of labelled reagent bound to the test and calibrator analytes.

Abstract: A method for measuring hyaluronic acid in a biological sample which comprises (a) coating a solid support with hyaluronic acid; (b) incubating the sample with cartilage proteoglycan; (c) exposing the incubated sample to the coated solid support; (d) then exposing the coated solid support to a keratan sulfate-reactive antibody; (e) determining the amount of antibody linked to keratan sulfate; and (f) correlating the amount of antibody linked to the keratan sulfate to the amount of hyaluronic acid in the sample.

Abstract: The present invention relates to a fermentation process for producing BMY-28960 and desxylosyl BMY-28960, and to a novel BMY-28960-producing organism belonging to the genus Actinomadura and designated as strain AB 1236 (ATCC 55208).

Abstract: A plasmid which contains a mammalian cell-derived autonomously replicating sequence DNA, a promoter and a gene for peptide production inclusive of the translation initiation codon.

Abstract: The present invention provides mutant proteins of steroid hormone receptors. These mutant proteins are useful in methods of distinguishing a steroid hormone receptor antagonist from a steroid hormone receptor agonist. The present invention also provides plasmids containing mutated steroid hormone receptor proteins and cells transfected with those plasmids. In addition, the present invention provides methods for determining whether a compound is a steroid hormone receptor antagonist or agonist. Also, the present invention provides methods of determining endogenous ligands for steroid hormone receptors.

Abstract: Processes for increasing antigen binding capacity of a major histocompatibility complex (MHC) molecule or a chain thereof are described. Such processes comprise treating an antigen-bound MHC molecule or a chain thereof with an effective amount of an unfolding agent to release undesired antigen from the MHC molecule or chain and then treating the antigen-free MHC molecule or chain with an effective amount of a refolding agent to produce a functional MHC molecule or chain. Additionally, compositions of MHC molecules that are substantially free of undesired antigens or that have been bound with desired specific antigens, alone or in combination with pharmaceutically acceptable carriers and/or cytotoxic agents, are described. The present invention has utility for therapeutic administrations, drug screening, and diagnostics.

Abstract: A method and reagents are provided for determining whether a human cell is a hemopoietic cell and whether a human tissue cell is in a neoplastic state. Human cells which express only leukocyte-plastin (l-plastin) are hemopoietic cells and human cells which express both l-plastin and tissue-plastin (t-plastin) are neoplastic. The method can be performed using isoform-specific plastin nucleotide probes or isoform-specific anti-plastin antibodies.

Abstract: As assay for the measurement of direct binding of a peptide that is a T-cell epitope to gene products of the major histocompatibility complex (MHC), classes I and II, on the surface of intact living antigen-presenting cells, is provided. The assay comprises incubating the labelled peptide with the cells, and monitoring the extent of binding by the addition of a probe that reacts with the ligand used to label the peptide. The assay is suitable for autoimmune diseases and other immunological disorders. Peptides having a sequence corresponding to a stretch of the sequence of the antigen relevant to an immunological disorder, or modifications thereof, which bind to gene products of MHC, but do not stimulate T-cells, are also useful for the treatment of said disorders.

Abstract: A dry reagent prepared by lyophilizing a labelled ligand-immobilized receptor complex or a labelled receptor-immobilized ligand complex is, on rehydration, useful for detecting analytes in samples in a variety of displacement assays.

Abstract: A method for assaying a medium for the presence of a substance that affects an SH2-phosphorylated ligand regulatory system. The method employs an SH2-like domain or a subdomain thereof and a phosphorylated ligand. The phosphophorylated ligand is capable of interacting with the SH2-like domain or a subdomain thereof to form an SH2-phosphorylated ligand complex. The SH2-like domain or subdomain and/or the phosphorylated ligand are present in a known concentration. The SH2-like domain or a subdomain thereof and the phosphorylated ligand are incubated with a substance which is suspected of affecting an SH2-phosphorylated ligand regulatory system. The method is carried out under conditions which permit the formation of the SH2-phosphorylated ligand complex. SH2-phosphorylated ligand complex, free SH2-like domain or subdomains thereof, or non-complexed phosphorylated ligand are assayed.

Abstract: This invention presents a non-isotopic competitive assay for Vitamin B12 (B12), utilizing intrinsic factor (IF) labelled with horse radish peroxidase (HRP), by coupling via heterobifunctional cross-linking agents. In addition, a method for stabilizing the resultant conjugates by pretreatment with N-ethylmaleimide is disclosed.

Abstract: An assay for screening snake venom for the presence or absence of platelet aggregation inhibitors (PAIs) based on specific receptor binding is described. Using this assay, the identification and characterization of the PAI in a wide range of snake venom samples were accomplished. The purified PAI from several of these active snake venoms is described. In addition, PAIs lacking the Arg-Gly-Asp adhesion sequence but containing Lys-Gly-Asp are prepared and shown to specifically inhibit the binding of fibrinogen or von Willebrand Factor to GP IIb-IIIa.

Abstract: Methods of monitoring and detecting the early onset of organ injury incident rejection of an organ rejection in an animal are disclosed. The described methods are capable of distinguishing organ rejection injury from other organ tissue damage in the animal. Free secretory component levels in an animal biological fluid (e.g., bile, urine, blood, amniotic fluid) may be used to identify organ rejection in an animal. Multiple and single organ transplant patients may be monitored and diagnosed according to the claimed methods. Biological fluids, such as blood, (serum) or urine, are analyzed immunologically using a particularly adapted ELISA which are then compared to an FSC control concentration to identify elevated FSC values. Animals with test FSC above FSC control concentrations are diagnosed as having an ongoing organ rejection episode. The detection of congenital renal dysfunction in utero is also provided according to the present invention through the measurement of FSC in the amniotic fluid.

Abstract: A monoclonal antibody recognizing an autocrine growth factor that reacts with activated lymphocytes and cancer cells is described. The growth factor, a small glycoprotein is distinct from interleukin 2 and other known growth factors by function, structure and tissue distribution. A homologous growth factor is present in lymphoid tissues of a wide range of vertebrate species. Diagnostic and therapeutic uses of the growth factor and antibody-growth factor complex are disclosed. Also disclosed are therapeutic uses of synthetic analogues and peptide derivatives of the antigen.

Abstract: An assay for detecting molecules and compounds which specifically bind to sites which regulate cellular potassium channels, for use, inter alia, as a method for identifying drugs with activity specific for modulating K.sup.+ channels.

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: This invention provides for improved means to produce binding specific proteins using concatamers of semi-randomly generated oligonucleotides inserted into genes encoding the external domains of bacterial outer membrane proteins. The genes are induced to express and the bacteria are then screened for the ability to bind to predetermined compositions. Those clones carrying the desired binding protein are isolated, cultured and the protein purified. Increased avidity of the binding specific proteins are achieved by reisolating the oligonucleotides which conferred binding affinity and mixing them with new semi-randomly generated oligonucleotides to generate a population enriched for oligonucleotides that had previously conferred to bacteria the desired binding affinity.

Abstract: Ligand reagents are disclosed which consist essentially of recombinant hyperglycosylated cytokines, expressed in yeast, which are purified and conjugated to various functional moieties, for example, biotin groups, via oligosaccharide residues. Methods of using such ligand reagents are also disclosed.

Abstract: Hybridization assay probes specific for Staphylococcus aureus and no other Staphylococcus species.

Abstract: This invention is directed to a composition comprising a substantially purified protein which enhances endogenous stimulus-induced neurotransmitter release, said neurotransmitter being selected from the group consisting of acetylcholine, dopamine or serotonin, said protein having a binding affinity constant of at least 2.9.+-.0.8 nM for tritiated 3,3-bis(4-pyridinylmethyl)-1-phenylindolin-2-one as determined by a Scatchard analysis, and said protein being trypsin sensitive and phospholipase C insensitive.

Abstract: A method disclosed for studying the interaction in solution of two molecules of the type such as a ligand and a receptor that are capable of reacting or binding with each other. The method comprises preparing an aliquot of a solution containing the first of the molecules. The second of the molecules is then added to the aliquot. A fluorescently labeled molecule is added to the aliquot, wherein the fluorescently labeled molecule is also capable of reacting or binding with the second of the molecules. A porous matrix that is optically transparent is immersed into the aliquot containing the two molecules being studied and the fluorescently labeled molecule, wherein the second molecule and any fluorescently labeled molecule bound thereto is sterically hindered from permeating the porous, optically transparent matrix, while any unbound fluorescently labeled molecule permeates the matrix.

Abstract: Alkaline phosphatase to which is covalently bound a carbohydrate of the general formula ##STR1## wherein n is 0, 1, 2 or 3, A is an acyl radical containing 2 to 5 carbon atoms, R.sup.1 and R.sup.2 are each hydrogen atoms or hydroxyl groups, R.sup.3 is --COOH or --CH.sub.2 OH, R.sup.4 is a hydroxyl group or a --CHOH--CHOH--CH.sub.2 OH or --NH--CO--CH.sub.2 --CH(NH.sub.2)--COOH radical or a complex containing the carbohydrate, is used as a standard for the determination of human alkaline phosphate. Preferred carbohydrates are ovomucoid, chitobiose, chitotriose or N-acetylglucosamine-N-acetylneuraminic acid.

Abstract: Method of determining, in a blood sample, containing unwashed platelets and fibrinogen, the state of thrombin reactivity of the platelets by adding thrombin to the sample in the presence of an agent for inhibiting fibrin polymerization, and then detecting activated platelets.

Abstract: A method for detecting the presence of a high-molecular-weight analyte, which uses a variation on complementation assays, comprising (1) providing an enzyme donor (ED)complex comprising ED coupled to an analyte-specific binding molecule, wherein the ED complex retains measurable complementation activity such that active .beta.-galactosidase is formed in the presence of an enzyme acceptor (EA); (2) contacting the ED complex and EA in the presence of a sample suspected of containing a high-molecular-weight analyte that reacts specifically with the analyte-specific binding molecule; and (3) relating the presence of the analyte in the sample to the formation of active .beta.-galactosidase enzyme.

Abstract: A diagnostic kit for detecting the presence of microorganisms, comprising an insoluble substrate; and a carbohydrate receptor immobilized on the insoluble substrate, the carbohydrate receptor being capable of adsorbing microorganisms; and a labelled reagent useful for detecting the presence of microorganisms bound to the carbohydrate receptors and a method for detecting the presence of specified microorganisms in a sample, which comprises contacting a sample to be tested with carbohydrate receptors immobilized on an insoluble substrate; and determining the extent of binding of microorganisms in the sample to the carbohydrate receptors by use of a labelled reagent.

Abstract: The present invention provides reagents for a detectable label for use in specific binding assays. Specifically, modified .beta.-galactosidase enzyme donors (ED) and acceptors (EA) are utilized. Both ED and EA are modified to form separate ED and EA complexes by coupling each with a linking element and a binding moiety which is specific to a binding site in the analyte. The ED and EA complexes are incapable of forming active enzyme in the absence of the analyte. However, when analyte is present, ED and EA complexes which bind to a common analyte in a sample, active .beta.-galactosidase is formed.

Abstract: Soluble HLA cross-matches are determined by providing for antibodies or ligand bound to a solid substrate specific for at least one HLA allele and detecting complexes between either donor or recipient HLA antigens and recipient or donor antibodies, respectively, or reference HLA and patient antibodies, particularly IgG antibodies. Conveniently, anti-human immunoglobulin (Ig), particularly human IgG, conjugates are employed where the anti-human Ig is conjugated with a label capable of providing a detectable signal to permit detection of human anti-HLA bound to said solid substrate.

Abstract: A method of binding aggregated immunoglobulin or immune complexes comprising contacting them with serum amyloid P component ("SAP"). The invention also comprises methods of using SAP to detect or quantitate immune complexes and to deplete fluids of aggregated immunoglobulin or immune complexes for diagnostic or therapeutic purposes. Also provided is a test kit comprising SAP for detecting or quantitating immune complexes and a device for removing aggregated immunoglobulin or immune complexes from fluids.

Abstract: The invention is directed to a method of purifying sequentially synthesized peptides and oligonucleotides by affinity techniques. Selected products are capped with and N-terminus capping agent for peptides or a 5'-terminus capping agents for oligonucleotides, and then bound with affinity agents that are selective for the corresponding capping agents.

Abstract: A replication initiator protein complex for eukaryotic cells is disclosed which includes protein fractions of about 60 kD and 100 kD size. The protein complex comprises a protein in purified form that is capable of origin-specific DNA binding and ATP-dependent DNA helicase activity. The 60 kD fraction is named RIP60 and is primarily active in origin-specific DNA binding. The 100 kD fraction is named RIP100 and is primarily active as a DNA helicase. The protein complex and its fractions are active only during S phase and thereby commend their investigation and use toward the development of specific diagnostic and therapeutic modalities against pathogens to which cell division is critical. Diagnostic and therapeutic utilities are accordingly proposed, and testing procedures, materials in kit form, recombinant materials and procedures, and pharmaceutical compositions are likewise set forth.

Abstract: A ligand-mediated immunofunctional assay (LIFA) method for detecting the presence and the concentration of polypeptide hormone binding proteins which comprises capturing the binding protein with a solid phase bound first antibody, saturating the bound hormone binding protein with the ligand polypeptide hormone, and detecting the bound ligand polypeptide hormone with a detectably labeled second antibody specific for the ligand polypeptide hormone. In the absence of added saturating polypeptide hormone, the LIFA measures the amount of hormone binding protein bound to the endogenous ligand polypeptide hormone. A growth hormone binding protein assay illustrates the method of the present invention.

Abstract: A method of identifying potential polypeptide vaccines to an agent, such as viruses, bacteria, and parasites. A critical binding segment of a first polypeptide known to bind to a first MHC type, is ascertained. The effect of replacing each of the amino acids in the critical segment, upon binding of that segment to the first MHC type, is evaluated. Following this, a protein produced by the agent is scanned for at least one trial amino acid sequence which the foregoing evaluation indicates will be a good binder to the first MHC type. When a potentially good binding sequence is found, a polypeptide containing such sequence can be evaluated as a synthetic vaccine.

Abstract: A method and kit are provided for determining levels in a biological sample of free IGF-I, IGF-II, or GH ligand that is normally associated in the sample with a binding protein. This method involves contacting the body fluid with an immobilized unlabeled capture reagent and incubating at 4.degree.-10.degree. C. for no greater than about 4 hours to bind the free ligand contained in the body fluid; separating the fluid from the immobilized capture reagent; and measuring the level of free ligand now bound to the capture reagent. This method is particularly useful to determine levels of free IGF-I in serum or plasma.

Abstract: The present invention concerns a method to catalyze reporter deposition to improve detection or quantitation of an analyte in a sample by amplifying the detector signal which comprises reacting an analyte dependent enzyme activation system with a conjugate consisting of a detectably labeled substrate specific for the enzyme system, said conjugate reacts with the analyte dependent enzyme activation system to form an activated conjugate which deposits substantially wherever receptor for the activated conjugate is immobilized, said receptor not being reactive with the analyte dependent enzyme activation system. In another embodiment the invention concerns an assay for detecting or quantitating the presence or absence of an analyte in a sample using catalyzed reporter deposition to amplify the reporter signal.

Abstract: The present invention relates to a fermentation process for producing BMY-28960 and desxylosyl BMY-28960, and to a novel BMY-28960-producing organism belonging to the genus Actinomadura and designated as strain AB 1236 (ATCC 55208).

Abstract: A method of purifying LFA-3 using affinity chromatography. Purified LFA-3 is useful for quantitating or separating out CD-2-containing cells.

Abstract: The invention provides the discovery that estrogen receptor is a constitutive transcriptional activator and a repressor. These activities are lacking in the mutant estrogen receptor. The invention provides assays and methods for determining estrogen binding activity of ligands. The invention also provides therapeutic methods and the detection of pathologies associated with a mutated estrogen receptor.

Abstract: Disclosed are compositions and methods for diagnosing autoimmune insulin dependent diabetes mellitus. In general, diagnostic methods of the invention include testing for the presence of an autoimmune immunoglobulin in a suspected patient's serum wherein the immunoglobulin is identified by its ability to interfere with the glucose transporting activity of a pancreatic islet cell glucose transporter. The inventors have discovered that the presence of such an autoimmune antibody in patient's serum is diagnostic of autoimmune insulin dependent diabetes mellitus. In particular aspects the diagnostic assay of the invention involves the incubation of isolated and dispersed islet cells, such as rat islet cells, in the presence of immunoglobulin obtained from the patient. Following such an incubation, the islet cells are tested for their ability to uptake glucose. A diagnosis is made through a determination that immunoglobulins in the serum of the patient specifically inhibit the uptake of glucose by the islet cells.

Abstract: A method for identifying compounds useful for treating patients with amyloidosis is disclosed. Compounds are screened according to the present invention to determine their ability to modulate the affinity between amyloid protein and proteins of the extracellular matrix.

Abstract: Methods are disclosed for conducting assays. One such method comprises providing in combination a first bibulous member zone ("first zone") and a liquid medium containing a component. The first zone has non-diffusively bound thereto a reagent interreactive with the component. Conditions are selected wherein the liquid medium and at least a portion of the component contained therein traverse all of the first zone and migrate by capillary migration into a second bibulous member zone ("second zone"). The second zone is of a different composition than the first zone and is incapable of specifically binding the component except when an analyte is to be detected and the method further includes causing a reagent to become bound to the first bibulous member zone in relation to the amount of analyte present.

Abstract: Methods are provided for the purification of interleukin-2 (IL-2) from a wide variety of sources, including synthetic mixtures, culture medium conditioned by natural IL-2 producing cells, and mammalian and bacterial recombinant IL-2 expression systems. The methods of the invention employ IL-2 receptor-affinity adsorbents in which soluble IL-2 receptors have been immobilized on solid supports. Through the use of these affinity adsorbents, highly purified IL-2 can be produced in a single step from bacterial extracts or conditioned medium. The IL-2 thus purified is largely free of aggregated forms, which are often present when other purification methods are used.

Abstract: Methods are provided for the purification of interleukin-2 (IL-2) and chimeric proteins containing an IL-2 moiety from a wide variety of sources, including synthetic mixtures, cell culture conditioned medium and mammalian and bacterial recombinant expression systems. The methods of the invention employ IL-2 receptor-affinity adsorbents in which soluble IL-2 receptors have been immobilized on solid supports. Through the use of these affinity adsorbents, highly purified IL-2 and chimeric proteins containing an IL-2 moiety can be produced in a single step from bacterial extracts or conditioned medium. The proteins thus purified are largely free of aggregated forms, which are often present when other purification methods are used.

Abstract: The detection of toxins through the use of an optical sensor having a fiber upon which is coupled one or more physiological receptors.