Abstract: Analytes can be detected from among closely related substances by reacting the analyte in a test sample with a labeled binding agent which specifically binds to the analyte to form a complex. The labeled binding agent is supplied in excess, and the complex is identified through a time window relative to the detection of the excess unbound agent. The complex and labeled binding agent are isolated on a separation media and identified by the differential rate of migration.

Abstract: A method for determining the complexed forms of immunologically determinable prostate specific antigen (cPSA) in a blood sample, e.g., by two-site immunometric assays, in which the blood sample is treated to render free PSA (fPSA)immunologically nondetectable. A particularly preferred immunometric assay method employs three anti-PSA antibodies: an antibody that binds to both cPSA and fPSA (anti-tPSA), a second anti-tPSA antibody which is characterized by the unique property that binding to fPSA is blocked by binding of fPSA-specific antibodies, and a third antibody which is a fPSA-specific antibody. Thus, binding of the fPSA-specific antibody to PSA in the sample allows only cPSA to be measured in the immunometric assay. Measurement of cPSA blood levels has been found to provide a method for aiding in the diagnosis and monitoring of prostate cancer that is highly sensitive and specific, and eliminates the need for a significant number of patients to undergo unnecessary prostate biopsy.

Abstract: The invention relates to polypeptides possessing urease activity of the type expressed naturally in C. pylori and immunogenic compositions comprising those polypeptides. This invention also relates to antibodies to polypeptides possessing urease activity and use of those antibodies to detect C. pylori.

Abstract: This invention relates to InhA enzyme crystals and to methods of growing said crystals. This invention is further directed to the utilization of said crystals to determine the three dimensional structure of InhA enzyme utilizing heavy atom derivatives of said crystals, and to the identification and development of compounds which inhibit the biochemical activity of InhA enzyme in bacteria and plants.

Abstract: Novel assay kits and methods are described for the detection and determination of the severity of disease conditions associated with elevated levels of aspartate aminotransferase (AST). The AST levels are determined by reaction of the AST with cysteine sulfinic acid (CSA) substrate in the presence of a sulfite-reactive dye compound that is substantially nonreactive with AST or CSA, such as a triarylmethine dye. Sulfite ion formed by the reaction of AST with CSA causes a calorimetric indication of AST that can be related to a predetermined stage of disease.

Abstract: Anti-human stromelysin monoclonal antibodies reactive with latent and active forms of stromelysin without discrimination between the two, each being immunoreactive with only one of the antigenic determinants of human stromelysin, are provided. The use of a combination of two such monoclonal antibodies which specifically react with different antigenic determinants of human stromelysin renders it possible to accurately determine the amount of human stromelysin in human body fluids, and thus to carry out the diagnosis of rheumatoid arthritis.There are thus provided said monoclonal antibodies per se, a sandwich enzyme immunoassay for the determination of the amount of human stromelysin in human body fluid samples using a combination of two such monoclonal antibodies, and a method for the diagnosis of rheumatoid arthritis using said immunoassay.

Abstract: A method for diagnosing chronic fatigue syndrome in an individual. Peripheral blood monocytes are isolated and p68 kinase activity, mRNA levels, protein levels, apoptosis and cell cycle analysis are measured. Significantly increased levels of any of these compared to healthy control individuals indicates the presence of chronic fatigue syndrome.

Abstract: A method and system for separating a selected molecule from a heterogeneous mixture of molecules in aqueous solution are described. The method comprises (a) providing a separation device comprising a loading reservoir and a receiving reservoir coupled by a channel bearing immobilized microtubules aligned parallel to the longitudinal axis thereof the channel; (b) placing an aqueous solution containing the heterogeneous mixture of molecules in the loading reservoir; (c) adding a motor-ligand composition and ATP to the aqueous solution, wherein the motor-ligand composition comprises a motor protein for attaching to microtubules and moving therealong in the presence of ATP and the ligand is capable of binding the selected molecule, such that the ligand binds the selected molecule to form a complex and the complex moves along the immobilized microtubules to the receiving reservoir; and (d) removing the selected molecule from the receiving chamber.

Abstract: The invention provides a method of detecting a class of substances, by providing an assay configuration which allows for reaction with a homologous portion common to all members of the class. The configuration utilizes a capture reactant to immobilize class members, revealing previously unrecognizable reaction sites on the class. The capture reactant interacts with a homologous portion on the class members. The immobilized class is then detected, such as by the use of a sandwich antibody directed to another homologous portion of the class now exposed for reaction, or directed to specific portions of specific members of the class that have been exposed for reaction. These reaction sites were unrecognizable prior to immobilization of the class members with the capture reactant.

Abstract: The invention relates to an enzyme linked immunosorbent assay (ELISA) kit for the rapid determination of N-acetyltransferase (NAT2) phenotype which can be used on a routine basis in a clinical laboratory. The ELISA kit allows physicians to: a) individualize therapy of drugs such as amrinone, procainamide, amonafide, dapsone, isoniazid, trimethoprim-sulphamethoxazole (TMP-SMX) and b) to predict susceptibility to carcinogen induced diseases such as bladder and colon rectal cancers.

Abstract: This invention provides a method for identifying a cellular protein capable of specifically binding to an activated antibody receptor, whose cytoplasmic domain comprising an ARH1 motif, comprising (a) obtaining cells comprising receptors having the ARH1 motif; (b) lysing the cells under conditions whereby the native complex of the receptor having the ARH1 motif and the cellular protein is preserved;(c) isolating the complex; and (d) testing the associated receptor and the protein for biochemical activities, thereby identifying the cellular protein capable of specifically binding to an activated antibody receptor, whose cytoplasmic domain comprising an ARH1 motif.

Abstract: A saccharide composition containing trehalulose, which is obtainable by allowing a maltose/trehalose converting enzyme to act on a sucrose solution to produce trehalulose, and collecting the resulting trehalulose-containing mixture. Since the enzyme converts sucrose into trehalulose in a relatively high yield and the conversion rate is controllable, a saccharide composition rich in trehalulose is readily obtained on an industrial scale.

Abstract: The invention provides isolated nucleic acid compounds encoding the glycosyltransferase protein GtfD of Amycolatopsis orientalis. Also provided are vectors carrying the gtfD gene, transformed heterologous host cells for expressing the GtfD protein, and methods for producing glycopeptide compounds using the cloned gtfD gene.

Abstract: The invention concerns monoclonal antibodies which bind to the CK-MB isoenzyme but not to the B or M subunit of CK-MB or to the CK-MM and CK-BB isoenzmyes, as well as a method for the diagnostic detection of CK-MB in a homogeneous diagnostic test using these antibodies.

Abstract: The present invention relates to a complex comprising nerve growth factor (NGF) and trk-proto-oncogene protein. The present invention also relates to methods for detecting the presence of NGF and trk-proto-oncogene receptor. The present invention further relates to methods that can be used in diagnostics and therapeutics for neurodegenerative diseases such as Alzheimer's and Huntington's by detecting NGF-trk receptor pairs and the phosphorylation of trk protein.

Abstract: The present invention provides antibodies capable of discriminating between native human MGMT and an active site alkylated form of this enzyme in an immunoprecipitation procedure.Monoclonal antibodies having such discriminating ability are obtainable from hybridoma ECACC 92112510 and hybridoma ECACC 93112514, which were deposited at the European Collection of Animal Cell Cultures, UK under the Budapest Treaty on 25th Nov. 1992 and 25th Nov. 1993 respectively. The monoclonal antibody of hybridoma ECACC 92112510 has been designated Mab 5H7. The monoclonal antibody of hybridoma ECACC 93112514 has been designated Mab 3C7.

Abstract: A process for isolating nucleic acids from agarose, and particularly regular agarose, is described wherein the agarose is augmented with a chaotropic substance and hydrolyzed by a novel purified agarase enzyme from Flavobacterium sp. strain NR19.

Abstract: Disclosed are methods of electrochemical analysis of an analyte in a sample. The methods include providing a sample comprising an analyte, and introducing the sample into a capillary tube containing electrophoretic running buffer and a reactant. According to the methods of the invention, one or both of the analyte and the reactant is electrically charged. Chemical contact between analyte and reactant induced by their relative electrophoretic mobility results in the breaking or formation of a covalent bond of one of the analyte or the reactant to produce a detectable product. The methods also include the steps of imposing along the length of the capillary tube an electric current for a time sufficient to bring into chemical contact within the tube analyte and reactant so as to allow formation of product, and detecting the product.

Abstract: The present invention provides compositions and methods useful for isolating calcineurin as well as inhibiting calcineurin activity. The compositions are peptides that contain regions that are homologous to calcineurin-binding regions of AKAP 79. Also provided are methods for determining if a cell contains a calcineurin-binding and PKA-binding anchoring protein that are useful for identifying additional proteins that bind both calcineurin and PKA.

Abstract: The present invention provides such a measuring method of the CK isoform and a reagent therefor. Accordingly, the present invention provides a reagent for measuring creatine kinase (CK) activity comprising an antibody, wherein said antibody inhibits CK-M.sub.T subunit, but does not inhibit CK-M.sub.S subunit.The present invention also provides a method for measuring CK activity comprising determining an inhibition of CK of body fluids using a reagent for measuring CK activity, wherein said reagent comprises an antibody which inhibits CK-M.sub.T subunit, but does not inhibit CK-M.sub.S subunit.

Abstract: Fluorescent-labeled substrates are provided for fluorescence polarization says of enzymes. These substrates are proteins labeled with derivatives of BODIPY.RTM., 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene. The BODIPY.RTM. fluorescent tag of the present invention is pH independent, and can be used over a pH range of from about 2 to about 11. Thus one can assay, in real time, enzymes with pH maxima at pH below 7 using fluorescence polarization methodology, which could not be done with fluorescein derivatives. Different enzymes can be compared using the same BODIPY.RTM. conjugate by merely changing the buffer system which changes the pH conditions.Fluorescence polarization assays of enzyme activity can be performed in the presence of whole bacteria and other finely suspended particles, such as might be present in tissue homogenates or cellular material. This is particularly useful for chairside assays on dental plaque or clinical assays on bacteria or tissue or exudates.

Abstract: The object of the invention is the cloning and sequencing of the nucleic acid coding for the human testicular ACE as well as the determination of the peptide sequence of this ACE.It relates more particularly to the use of nucleotide probes capable of hybridizing with all or part of the above-mentioned nucleic acid, and also the polyclonal or monoclonal antibodies recognizing specifically all or part of the above-mentioned peptide sequence, and the use of these probes or these antibodies for the implementation, respectively, of a method for the detection of the messenger RNA coding for the testicular ACE in vitro or of the testicular ACE directly.

Abstract: A method is provided for determining the presence and/or amount of a microorganism and/or its intracellular material present in a sample performed by: (a) exposing the sample to a specific binding agent that has been immobilized upon a solid substrate, the specific binding agent being capable of binding to the microorganism or its intracellular material such that it becomes associated with the solid substrate, (b) exposing the solid substrate to an agent capable of making adenylate kinase associated with the microorganism and/or its intracellular material accessible to solutions applied to the substrate, (c) applying a solution containing adenosine diphosphate (ADP) to the substrate under conditions whereby adenosine triphosphate (ATP) may be produced by any adenylate kinase present, and (d) measuring the amount of adenosine triphosphate (ATP) and relating that to the presence and/or amount of microorganism or intracellular contents.

Abstract: The present invention provides monoclonal antibodies against peptides having an amino acid sequence described in Sequence No. 1 or No. 2, the peptides being found in human thymidine phosphorylase and human platelet-derived endothelial cell growth factor. The Invention also provides an immunoassay for human thymidine phosphorylase and/or human platelet-derived endothelial cell growth factor using the monoclonal antibodies. The monoclonal antibodies of the invention recognize human thymidine phosphorylase and human platelet-derived endothelial cell growth factor, and thus are useful in the diagnosis and treatment of various tumors and their metastasis and diseases accompanying abnormal angiogenesis.

Abstract: A method of assaying for CAT in a fluid involves the use of a complex of chloramphenicol with a member of a specific binding pair such as a hapten or biotin. Biotinylated chloramphenicol is claimed as new. A scintillation proximity assay involves use of this reagent with tritiated acetyl coenzyme A and streptavidin coated SPA beads.

Abstract: This invention discloses novel immunoassay termed an ELONA, employing nucleic acid ligands as capture molecules and/or detector molecules.

Abstract: The invention provides methods for detecting elevated levels of phospholipase A.sub.2 activating protein in persons suspected of having rheumatoid arthritis to thereby indicate the presence of rheumatoid arthritis in the person comprising the steps of providing a sample of body fluid or tissue from said person; contacting the sample with an antibody specific for phospholipase A.sub.2 activating protein such that the antibody binds with phospholipase A.sub.2 activating protein in the sample; detecting the antibody thereby indicating the presence of phospholipase A.sub.2 activating protein, whereby elevated levels of phospholipase A.sub.2 activating protein in the sample as compared with levels found in persons not having rheumatoid arthritis indicates the presence of rheumatoid arthritis in the person. Kits and reagents for detecting rheumatoid arthritis are also provided.

Abstract: A method is provided for preparing lipoprotein (a) from a volume of biological fluid that is substantially free of lipoproteins of another class. The method involves an ultracentrifugation step in which at least one fraction is recovered that contains Lp(a). This material is then reacted with immobilized ligand to remove non-Lp(a) interfering substances from the fraction, the Lp(a) remaining unbound. The non-reacted Lp(a) is subsequently obtained in a form that is suitable for use in the analysis of any of protein concentration, protein isoform determination or cholesterol assays. A method of identifying and measuring an amount of one or more isoforms of Lp(a) is further provided.

Abstract: Two or more analytes having the same action, or having different actions in spite of their similar structures, or two or more analytes having the same action and the same detectable chemical characteristics, in samples derived from living bodies, etc., can be measured rapidly and easily by forming one or more complexes with one or more affinity substances, separating the complexes by using high pressure liquid chromatography, followed by measurement of the amount of an affinity substance or one of the analytes.

Abstract: A method and apparatus for immunoassays utilizes an improved collection technique of fluorescence induced emissions at the solid phase/liquid phase interface from surface plasmon resonance sensing devices. In a preferred embodiment, a solid phase substrate is coated with a thin film of a conducting material on which a first specific binding partner is directly or indirectly immobilized. The coated solid phase substrate is incubated with a liquid component comprised of a biological sample containing a specific ligand or analyte and a fluorescent labeled second specific binding partner in the case of immunometric assays, or a fluorescent labeled ligand or analog thereof in the case of competitive assays.

Abstract: The subject invention pertains to a novel assay device for detecting oxalate in a sample. The assay device comprises enzyme and dye compositions immobilized on a solid carrier matrix. The subject invention can also be used to measure the concentration of oxalate in a sample. The subject invention further pertains to a novel oxalate oxidase composition and methods of preparing the subject enzyme composition. The oxalate oxidase composition can be used in the assay device of the subject invention.

Abstract: A method of treating a subject that is undergoing methylphenidate therapy and concomitant therapy with another drug undergoes or interferes with P.sub.450 metabolism, wherein the methylphenidate is d-threo-methylphenidate.

Abstract: Large numbers of bioactive compounds of great chemical diversity may be rapidly prepared by subjecting organic compound mixtures to a plasma discharge, followed by screening for specific bioactivity.

Abstract: Disclosed is a process of enzymatically preparing .alpha.-glucoside esters. First, .alpha.-glucosides are produced by placing an acyclic alcohol or a mixture of acyclic alcohols having a water solubility of at least 2.7% v/v at 20.degree. C. in contact with starch, maltodextrins or maltose, in the presence of a purified enzymatic preparation having .alpha.-transglucosylation activity, wherein the enzymatic preparation is free of .beta.-glucosidase activity. Then, the .alpha.-glucosides are contacted with at least one fatty acid and a preparation having lipase activity to produce the .alpha.-glucoside esters, which may then be recovered. The preparation having .alpha.-transglucosylation activity may originate from a fungus such as Talaromyces duponti, Aspergillus niger, Aspergillus oryzae or Aspergillus batatae.

Abstract: Purified N-acetylglucosaminidase and .alpha.1-3,6 Galactosidase endogenous to Xanthomonas have been described. Substrate specificity of isolated enzymes have been identified from GlcNAc.beta.1-x and Gal.alpha.1-3R, Gal.alpha.1-6R, providing improved capability for selectively cleaving a glycosidic linkage in a carbohydrate substrate and for forming modified carbohydrates.

Abstract: A method for detecting autoantibodies to glutamic acid decarboxylase (GAD) in the serum of a patient as diagnostic of a diabetic or prediabetic condition in the patient, comprises contacting a serum sample from the patient with a GAD antigen and detecting binding of autoantibodies to GAD in the sample by the GAD antigen, wherein the GAD antigen comprises a GAD preparation containing an enhanced amount of dimer(s) or oligomer(s) of the 65 kD or 67 kD isoforms, or both, of GAD. A diagnostic kit is also inclosed.

Abstract: The invention provides a method of increasing the sensitivity of a cell to a DNA damaging agent comprising increasing the amount of an NM23 in the cell. The method is especially useful for increasing the sensitivity of tumor cells to chemotherapy and radiation.

Abstract: The present invention relates to an enzyme with .beta.-(1-6)-endoglucanase activity encoded by a DNA sequence, which DNA sequence a) comprises the DNA sequence shown in SEQ ID No. 3, or b) comprises an analogue of the DNA sequence shown in SEQ ID No. 3, which i) is homologous with the DNA sequences shown in or SEQ ID No. 3, and/or ii) hybridizes with the same oligonucleotide probe as the DNA sequence shown in SEQ ID No. 3, and/or iii) encodes a polypeptide which is homologous with the polypeptide encoded by a DNA sequence comprising the DNA sequence shown in SEQ ID No. 3, and/or iv) encodes a polypeptide which is immunologically reactive with an antibody raised against a purified .beta.-(1-6)-glucanase shown in SEQ ID No. 4 derived from Trichoderma harzianum, CBS 243.71.

Abstract: Regioselective acetylation of the 9-hydroxyl group on N-acetylneuraminic acid is achieved enzymatically for producing oligosaccharides which contain a terminal N-acetylneuraminic acid moiety. This method provides access to O-acylated disialogangliosides as well as other N-acetyl-neuraminic acid oligosaccharides. These compounds are biologically and medicinally important and are difficult to obtain from nature or by chemical acylations. The methodology affords simple reaction conditions and minimal purification steps. In addition, the process affords good yields and the enzymes and reagents employed are commercially available with high stability and low costs.

Abstract: The method of measuring analytes in biological fluids is disclosed wherein a specific binder to a given analyte is covalently immobilized onto a solid support to which a labeled analyte is pre-reacted and stabilized to form a binder-labeled analyte complex. A sample is contacted with said immobilized complex wherein an analyte in the sample, if present, competes with the labeled analyte bound to the immobilized binder for binding sites on said binder thus displacing a given amount of the labeled analyte which is directly proportional to the amount of analyte present in the sample. The affinity of the labeled analyte to the analyte's specific binder is lower than the affinity of the unlabeled analyte to the same binder.

Abstract: An assay for measuring activation (i.e., autophosphorylation) of a tyrosine kinase receptor of interest is disclosed.(a) A first solid phase is coated with a substantially homogeneous population of cells so that the cells adhere to the first solid phase. The cells have either an endogenous tyrosine kinase receptor or have been transformed with DNA encoding a receptor or "receptor construct" and the DNA has been expressed so that the receptor or receptor construct is presented in the cell membranes of the cells.(b) A ligand is then added to the solid phase having the adhering cells, such that the tyrosine kinase receptor is exposed to the ligand.(c) Following exposure to the ligand, the adherent cells are solubilized, thereby releasing cell lysate.(d) A second solid phase is coated with a capture agent which binds specifically to the tyrosine kinase receptor, or, in the case of a receptor construct, to the flag polypeptide.

Abstract: A method of screening for a given enzyme in colony-forming cells using a substrate of the enzyme that has low solubility. Use of such a method leads to the discovery of glutaryl-7-aminocephalosporanic acid (GL-7-ACA) acylase of Pseudomonas nitroreducens. Also disclosed is a method of obtaining GL-7-ACA acylase from Pseudomonas cells.

Abstract: A method for screening or detection of a non-enzyme catalytic polypeptide or protein for the conversion of a substrate S to a product P is provided, in which a preparation containing the potential catalyst is contacted with the substrate S immobilized to a support and the immobilized product P obtained is detected, preferably by immunoassay. The method is preferably used for the screening of hybridoma supernatants for catalytic monoclonal antibodies that catalyze acyl transfer reactions, e.g., hydrolysis or aminolysis, condensation reactions and resolution of enantiomers.

Abstract: The complete amino acid and nucleotide sequence for adenylate cyclase stimulating factor is provided, thereby facilitating the synthesis of ACSF in recombinant cell culture. ACSF amino acid sequence variants and ACSF antibodies are provided which are useful in the treatment of humoral hypercalcemia of malignancy or in immunoassays for ACSF. In particular, antibodies capable of binding only the C-terminal domains of ACSF are useful in immunoassays for ACSF which avoid interference by parathyroid hormone. Also provided are novel polypeptides selected from the group of the ACSF basic peptide, the ACSF C-terminal peptide, or the ACSF domain containing both of the basic and C-terminal peptides.

Abstract: Oligosaccharide compounds that are substrates and inhibitors of glycosyltransferase and glycosidase enzymes and compositions containing such compounds are disclosed. A method of glycosylation is also disclosed. An E. coli transformed with phagemid CMPSIL-1, which phagemid comprises a gene for a modified CMP-sialic acid synthetase enzyme, which transformed E. coli has the ATCC accession No. 68531 is also provided.

Abstract: The invention provides novel methods and compositions for detecting kinase activity in solution, without the use of radioactivity. The methods marry the kinetic advantages of solution-based reaction with the efficiency and high-throughput adaptability of solid-phase wash and detection steps, yet is conveniently practiced in a single tube. The methods may be used to assay for kinase activity per se or, by controlling for the kinase activity, for modulators of kinase activity. The method is exemplified with a preferred chemiluminescent protein kinase assay using biotinylated substrate peptides captured on a streptavidin coated microtiter plate and monoclonal antibodies to detect their phosphorylation.

Abstract: A method for producing monosialoganglioside GM1 comprising the step of contacting a crude ganglioside mixture with a microorganism capable of producing a sialidase; and a bacterial strain of Pseudomonas genus capable of producing a sialidase and usable in the above method.

Abstract: Protein kinase mutant and wild-type genes encoding polypeptides of the class heretofore designated "casein kinase I" and useful in screening compositions which may affect DNA double-strand break repair activity are disclosed. Also disclosed are methods using the polynucleotides in cell-proliferative disorders.

Abstract: A chemiluminescent assays for the determination of the presence or amount of a biopolymer in bound assays using 1,2-dioxetanes in connection with AttoPhos.TM. as chemiluminescent substrates for enzyme-labeled targets or probes is provided. Further disclosed is a kit for conducting a bioassay for the presence or concentration of a biopolymer comprising a) an enzyme complex; b) a 1,2-dioxetane; and c) AttoPhos.TM..

Abstract: A stable single reagent for the determination of carbon dioxide in serum is provided as an aqueous solution of diagnostically amounts and NADH and/or NADPH, and a stabilizing system therefore reactively stabilized PEPC and a substrate for reacting with bicarbonate ion under alkaline conditions in the presence of PEPC.