Abstract: Novel compounds comprising a C—C double bond substituted at one carbon with two sulfur atom-containing groups are disclosed. The compounds are useful in methods and compositions for generating chemiluminescence rapidly by reaction with a peroxidase enzyme and a peroxide. The chemiluminescence thus produced can be used as a detectable signal in assays for peroxidase enzymes or peroxide-producing enzymes and in assays employing enzyme-labeled specific binding pairs.

Abstract: The present invention provides novel glutathione-dependent formaldehyde dehydrogenase that makes possible quantitative measurement of formaldehyde by cycling reaction, and a determination method of formaldehyde and biological components, such as creatinine, creatine, and homocysteine, which produces formaldehyde as a reaction intermediate. In addition, the present invention provides a reagent kit for the above-mentioned determination method. The present invention provides a novel determination method of a homocysteine using transferase utilizing homocysteine and other compound as a pair of substrates. Particularly, the present invention provides a determination method of homocysteine which includes bringing betaine-homocysteine methyltransferase and dimethylglycine oxidase into contact with a sample and measuring produced hydrogen peroxide or formaldehyde. Moreover, the present invention provides novel dimethylglycine oxidase stable to thiol compound, which is suitably used for the measurement.

Abstract: A method for determining the presence of an analyte in a fluid is described along with various components of an apparatus specifically designed to carry out the method. The method involves taking a reflectance reading from one surface of an inert porous matrix impregnated with a reagent that will interact with the analyte to produce a light-absorbing reaction product when the fluid being analyzed is applied to another surface and migrates through the matrix to the surface being read. Reflectance measurements are made at two separate wavelengths in order to eliminate interferences, and a timing circuit is triggered by an initial decrease in reflectance by the wetting of the surface whose reflectance is being measured by the fluid which passes through the inert matrix. The method and apparatus are particularly suitable for the measurement of glucose levels in blood without requiring separation of red blood cells from serum or plasma.

Abstract: Novel compounds comprising a C—C double bond substituted at one carbon with two sulfur atom-containing groups are disclosed. The compounds are useful in methods and compositions for generating chemiluminescence rapidly by reaction with a peroxidase enzyme and a peroxide. The chemiluminescence thus produced can be used as a detectable signal in assays for peroxidase enzymes or peroxide-producing enzymes and in assays employing enzyme-labeled specific binding pairs.

Abstract: A dual enzyme chemiluminescent substrate formulation useful for the independent or simultaneous analysis of the enzymes is disclosed, the formulation includes, as one substrate component, dihydrophthalazinedione, such as luminol or isoluminol, and a peroxide source and, as the other substrate component, a 1,2-dioxetane. The formulation further includes a polymeric quaternary onium salt as an isolating agent to defeat adverse interactions between the substrate pairs.

Abstract: The object of the present invention is to provide a diagnostic method by which Helicobacter pylori infection can be diagnosed at low cost without causing pain on subjects and without requiring particular equipment and which is free of cross reactivity and excellent in specificity without variation among lots as a result of the use of a single antibody facilitating the quality control and which shows good sensitivity even when a single monoclonal antibody is used. The present invention provides a monoclonal antibody which recognizes Helicobacter pylori catalase as an antigen.

Abstract: A process for producing aldehydes, and/or carboxylic acids is described, in which a primary alcohol, especially a carbohydrate, is oxidized using a catalytic amount of a nitrosonium compound obtained by oxidizing a nitroxyl compound in the presence of an enzyme compound capable of oxidation. Further described are oxidized carbohydrates containing at least 1 cyclic monosaccharide chain group carrying a carbaldehyde group per 25 monosaccharide units and per molecule.

Abstract: The present invention relates to a method for conveniently quantitating TGs contained in various lipoproteins. A method for quantitating trigyceride (TG) in a particular lipoprotein which comprises eliminating free glycerol from a sample containing free glycerol and TG in the particular lipoprotein, then allowing the resulting sample to react with lipoprotein lipase and an enzyme system which generates hydrogen peroxide from free glycerol, and quantitating the generated hydrogen peroxide, is provided.

Abstract: The invention relates to nucleotide sequences coding for the accDA gene and to a process for the preparation of L-amino acids, especially L-lysine, by fermentation using corynebacteria in which the accDA gene is amplified.

Abstract: The present invention relates to a method for measuring a holoceruloplasmin concentration, and more particularly, to a method for measuring a holoceruloplasmin in a blood spot based on a standard concentration curve obtained through an enzyme-linked immunosorbent assay (ELISA) or a dissociation-enhanced time-resolved fluoroimmunoassay using a holoceruloplasmin-specific polyclonal antibody and a holoceruloplasmin-specific monoclonal antibody.

Abstract: A lateral flow chromatographic assay format for the performance of rapid enzyme-driven assays is described. A combination of components necessary to elicit a specific enzyme reaction, which are either absent from the intended sample or insufficiently present therein to permit completion of the desired reaction, are predeposited as substrate in dry form together with ingredients necessary to produce a desired color upon occurrence of the desired reaction. The strip is equipped with a sample pad placed ahead of the substrate deposit in the flowstream, to which liquid sample is applied. The sample flows from the sample pad into the substrate zone where it immediately reconstitutes the dried ingredients while also intimately mixing with them and reacting with them at the fluid front. The fluid front moves rapidly into the final “read zone” wherein the color developed is read against predetermined color standards for the desired reaction. Pretreatment pads for the sample, as needed, (e.g.

Abstract: According to the present invention, the biological or pharmacological activity of a test material like a plant or herbal material, an extract of a plant or herbal material, a natural or synthetic compound or some combination thereof, can be quantified by observing the pattern of structural changes induced in a eukaryotic cell's proteins. These structural changes may be evidenced by protein phosphorylation, by protein-protein interactions and the like. The amount and nature of protein phosphorylation is qualitatively and quantitatively related to the in vitro concentration of biologically/pharmacologically active components to which the mammalian cells are exposed. Additionally, formation or loss of protein-protein complexes may be determined in whole cell homogenates through the use of non-denaturing electrophoresis and staining for proteins or protein phosphorylation.

Abstract: The present invention relates to the human cellular protein glutathione peroxidase-gastrointestinal as a target for medical intervention against Hepatitis C virus (HCV) infections. Furthermore, the present invention relates to a method for the detection of compounds useful for prophylaxis and/or treatment of Hepatitis C virus infections and a method for detecting Hepatitis C virus infections in an individual or in cells. Also compositions, compounds, nucleic acid molecules (such as aptamers), mono- or polyclonal antibodies are disclosed which are effective for the treatment of HCV infections, and methods for prophylaxis and/or treatment of Hepatitis C virus infections or for the regulation of Hepatitis C virus production are disclosed.

Abstract: The present invention provides a method of detecting a peroxide-based explosive in a sample suspected of consisting of or comprising such explosive, which method comprises dissolving said sample in a suitable organic solvent, contacting the solution with an aqueous solution of a strong acid capable of decomposing said explosive to release hydrogen peroxide, and contacting the resulting mixture with a peroxidase enzyme. The invention also provides a kit for use in the method of the invention.

Abstract: An assay for detecting inhibitors of the enzyme myeloperoxidase said method comprising: reacting myeloperoxidase with hydrogen peroxide and a chloride source to generate hypochlorus acid in the presence of a potential inhibitor of said myeloperoxidase; reacting any formed hypochlorous acid with a suitable amine to form the corresponding chloramine; Optionally removing any unreacted hydrogen peroxide; reacting any formed chloramine with a suitable detector molecule in the presence of iodide to form oxidised detector molecule; determining the amount of formed oxidised detector molecule by measuring the absorbance or fluorescence at a suitable wavelength; Identifying as inhibitors of myeloperoxidase those compounds which cause the amount of formed oxidised detector molecule to be decreased. And for use in diagnostic tests for myeloperoxidase activity is disclosed.

Abstract: The invention provides chemiluminescent assays that incorporate a film including at least one chemiluminescent precursor immobilized therewith which produces a triggerable chemiluminescent compound, the film being free of compounds which generate singlet oxygen and being adapted for use with a sensitizer-labeled agent or agent probative of the analyte.

Abstract: An enzymatic assay for chlorite, chlorate and perchlorate determination based on horseradish peroxidase, chlorate reductase, or nitrate reductase and a reacting dye was developed. The assay is quick and simple requiring less than 30 min; is cheap costing less than $0.05 per sample; uses standard laboratory equipment; is sensitive to a lower limit of 0.4 micromolar and linear up to 6.0 millimolar; is highly reproducible and interference free. The application of this assay to chlorite analysis in treated water will significantly reduce one of the major costs associated with the use of chlorine dioxide as a disinfectant and improve the drinking water quality.

Abstract: One aspect of the present invention is a reagent test strip that includes: a) a top support layer including a sample aperture; b) a membrane that includes a reagent system for indicating the concentration of a target; c) a spreading layer; d) a bottom support layer including a measuring port in substantial alignment or approximate alignment with the sample aperture. Preferably, the membrane is affixed to the top support layer. Preferably, the membrane is positioned between the top support layer and the bottom support layer. Preferably, the spreading layer is positioned above the top support layer and in substantial or approximate alignment with the sample aperture. Preferably, a fluid applied to the spreading layer, passes through the sample aperture and contacts the membrane.

Abstract: Compositions, reagent test strips, analyte detection systems and kits of the same, as well as methods for their use in the detection of an analyte in a sample, are provided. The subject compositions are characterized by having a positively charged porous matrix and a urea derivative dye on at least one surface of the matrix, where in many preferred embodiments the urea derivative dye is a negatively charged urea derivative dye. In many preferred embodiments, the subject compositions further include at least one additional reagent member of a peroxide producing signal producing system, e.g., an analyte oxidase and/or a peroxidase. The subject compositions, test strips, analyte detection systems and kits find use in the detection of a wide variety of analytes in a sample, such as a physiological sample, e.g., blood or a fraction thereof.

Abstract: A method for counting leukocytes which comprises liberating elastase from granulocytes contained in a specimen, adding an anti-granulocyte elastase antibody to the thus liberated elastase, measuring the antibody bonded to the elastase to thereby determine the concentration of the elastase, and then calculating therefrom the number of leukocytes contained in the specimen with the use of the ratio of the leukocyte count to the elastase concentration. This method makes it possible to conveniently and less expensively count leukocytes at a high accuracy comparable to the one established by using a conventional automatic blood cell counter, as the figure shows.

Abstract: The invention relates to assays for measuring ubiquitin ligase activity and for identifying modulators of ubiquitin ligase enzymes.

Abstract: The invention relates to assays for measuring ubiquitin ligase activity and for identifying modulators of ubiquitin ligase enzymes.

Abstract: The invention features methods of detecting enzymatic activity (e.g., in a magnetic resonance image). In general, the methods include: (1) providing a monomeric substrate (e.g., a substrate that is polymerizable in the presence of an enzyme or as a result of an enzyme-catalyzed reaction), having the generic structure X-Y-Z, where X includes a chelator moiety having a chelated paramagnetic or superparamagnetic metal atom or ion, Y includes a linker moiety (e.g., to provide a covalent or non-covalent chemical bond or bonds between X and Z), and Z includes a polymerizing moiety; (2) contacting the substrate with a target tissue, wherein the substrate undergoes polymerization to form a paramagnetic or superparamagnetic polymer, the polymerization being catalyzed by an enzyme in an extracellular matrix or bound to the surfaces of cells of the target tissue; and (3) detecting an increase in relaxivity for the polymer relative to an equivalent amount of unpolymerized substrate.

Abstract: The present invention relates to an alcohol concentration assay test system, including compositions and methods for storing multiple alcohol concentration assay tests and compositions and methods for measuring the concentration of alcohol in an assay test sample. In particular, the present invention provides an alcohol assay test for use by individuals to monitor alcohol levels and a delivery system for alcohol assay tests.

Abstract: An immunoassay, e.g. ELISA, method and kit for determining (preferably quantitatively) an analyte adsorbed at a surface or present in a liquid sample, comprising binding the analyte to a solid phase, attaching a marker to the analyte, and detecting marker attached to the solid-phase. The invention proposes to use a combination of marker and detection (e.g. an enzyme-substrate combination) which is capable of producing a precipitate on a solid phase which carries the marker and to detect the binding of analyte to the solid phase by in-situ determining the change in surface mass of the solid phase due to the formation of the precipitate. Ellipsometry is an example of a technique suitable for determining the change of surface mass of the solid phase, which could be made of a silicon- or chromium-sputtered glass slide The invention shortens the assay time and/or improves the assay sensitivity, and allows to measure extremely low surface concentrations of analytes of interest.

Abstract: A process for oxidizing cellulose, in which a nitroxyl compound such as TEMPO is oxidized using an oxidizing agent in the presence of a complex of a transition metal such as Mn, Fe, Cu, and a complexing agent such as a polyamine, or an oxidative enzyme, and the resulting nitrosonium ion is used to selectively oxidize cellulose 6-hydroxy-methylene groups to carbaldehyde groups and carboxylic acid groups.

Abstract: A process for the on-line determination of the hydrogen peroxide content of a mixture obtained in a chemical reaction comprises at least the following steps:

Abstract: A single reagent system and a method to detect and measure oxidizing adulterants in bodily fluid being screened for drugs of abuse are disclosed. The system comprising a strip containing 0.05 to 0.2 micromole/25 sq. mm. of a benzidine derivative and is used to detect sodium hypochlorite (bleach), chlorine, hydrogen peroxide, sodium bromide, sodium iodide, sodium nitrite, and pyridinium chlorochromate adulterants in urine, sweat, saliva, blood or other bodily fluids during screening for drugs of abuse.

Abstract: A precursor for the construction of chelated metal conjugates which demonstrate improved assay performance and utility in minimizing non-specific binding while maintaining specificity for target molecules is disclosed. The precursor has tridentate functionality towards multivalent ions such as iron and nickel and contains a diacetyl glycine group covalently linked via an amide to a molecule such as a proteinaceous molecule providing a primary amide group for amide bond formation. The precursor is preferably prepared in monomeric form by reacting nitrilotriacetic acid or a salt thereof in an aqueous medium at an alkaline pH of at least 8 with a proteinaceous molecule containing a primary amine group in the presence of a carbodiimide. The proteinaceous molecule may be bovine serum albumin or an enzyme such as alkaline phosphatase or horseradish peroxidase.

Abstract: Microfluidic devices with improved channel and reservoir configurations are provided. More specifically, efficient microfluidic devices for the performance of temperature mediated reactions are provided. These reactions can be performed in an ease of use and efficient manner so as to enable high through put of multiple samples. Methods for performing temperature mediated reactions using the microfluidic devices with improved channel and reservoir configurations have also been provided.

Abstract: A washing solution for solid-phase immunometric methods which contains stabilizers for the labeling system, and the use thereof

Abstract: A tube or pipette-type element, such as a capillary tube or other tube element carries a drug identification system for a number of hypnotic-inducing drugs or anesthetic drugs. The drug identification system provides a visual indication of at least two or more drugs present in a liquid sample. The liquid sample is drawn into one end of the tube, moves along the tube, and functional materials and/or structure in the interior of the tube provide a visual indication of the presence of targeted contaminants (e.g., drugs) in the fluid sample. The design also minimizes or prevents any reflux of materials in the tube element back into the liquid or beverage from which the sample has been taken. An additional benefit of the system is safe disposal and moderate unit cost to the public.

Abstract: A method and apparatus for determining the hydrogen peroxide content of a fluid. The method includes: (a) contacting the fluid with a catalyst so as to permit decomposition of the hydrogen peroxide present in the fluid to oxygen and water; permitting the oxygen liberated to pass to gas meter; and (c) measuring the volume of oxygen liberated utilizing the gas meter, wherein the volume of oxygen liberated provides a measure of the hydrogen peroxide content of the fluid.

Abstract: The present invention involves a method for assaying a substance. The method of the present invention comprises contacting the substance with an assay agent comprising a catalytic agent to associate the substance with the catalytic agent, contacting the resulting associated substance with a label precursor capable of reacting catalytically with the catalytic agent to release the label, and detecting a mass label. The present invention also involves a kit for assaying a substance. The kit of the present invention comprises an assay agent comprising a catalytic agent, and a label precursor capable of reacting catalytically with the catalytic agent to release a mass label.

Abstract: The present invention provides internally calibrated competitive assays for use on a solid support. Additionally, the invention provides a method of using such assays.

Abstract: A process for measuring enzymatic activity in an identified, isolated, intact, single, viable cell. Each of the viable cells is placed within individual identified locations on a carrier of a cytometer having means to measure enzymatic activity of a single viable cell placed in an identified location. The identified isolated cell is exposed to a substrate of an enzyme to be measured, and the rate of product formed or released following every exposure of the cell to same or different concentrations of the substrate is measured. The isolated cell may be exposed to a sequence of at least two different concentrations of the substrate, and for each exposure the rate of product formed or released, is measured.

Abstract: A method for determining exposure of a test sample to hydrogen peroxide in an atmosphere includes placing a substrate into the atmosphere, contacting the substrate with the hydrogen peroxide, reacting the hydrogen peroxide with a dye intermediate to produce a chromophore indicative of the integrated exposure of hydrogen peroxide in the atmosphere; and correlating the chromophore to the integrated exposure of hydrogen peroxide in the atmosphere. The dye intermediate may be a dye-couple of 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH) and 3-dimethylaminobenzoic acid (DMAB). The intensity of the chromophore is indicative of an integrated hydrogen peroxide exposure.

Abstract: Provided is a method of screening gene libraries derived from a mixed population of organisms for a bioactivity or biomolecule of interest. The mixed population of organisms can be a cultured population or an uncultured population from, for example, the environment. Also provided are methods of screening isolates or enriched populations of organisms, which isolates include a population that is spatially, temporally, or hierarchical, for example, of a particular species, genus, family, or class of organisms. Identified clones containing a biomolecule or bioactivity of interest can be further variegated or the DNA contained in the clone can be variegated to create novel biomolecules or bioactivities of interest.

Abstract: This invention relates to the detection of patients at risk for developing integrin antagonist/agonist mediated disease states. This invention relates to assays useful for the detection in a patient bodily fluid sample of drug-dependent antibodies which bind to integrins, or intergrin-associated proteins or complexes thereof in the presence of an integrin antagonist/agonist. This invention also relates to assays useful for the detection in a patient bodily fluid sample of drug-dependent antibodies (DDABS) that bind to integrins, including the platelet glycoprotein IIb/IIIa (GPIIb/IIIa), in the presence of a integrin agonist and/or antagonist. This invention also relates to procedures for identifying integrin antagonists/agonists that are less prone to elicit integrin antagonist/agonist mediated disease states. This invention also relates to procedures which increase the recovery of integrin-directed antibodies in body fluids, resulting in an increased sensitivity and specificity of DDAB detection assays.

Abstract: A method for enhancing the conversion of a phenol substrate to a product by a peroxidase enzyme comprises the steps of reacting a conjugate comprising a detectably labeled phenol-containing molecule with a peroxidase enzyme in the presence of an enhancing reagent, the enhancing reagent comprising an organic compound having the structure wherein each R is independently selected from the group consisting of: hydrogen and a C1-12 substituent; where V, W, Y and Z are each independently selected from the group consisting of: H, halogen, the C1-12 substituent, NR2, OR and SR; and where X is selected from the group consisting of: H, Br, Cl, F, the C1-12 substituent, NR2, OR and SR; or synergistic mixtures of an inorganic salt and the organic compound. The C1-12 substituent is linear, branched or cyclic. The C1-12 substituent is alkyl, alkenyl, alkynyl, heteroatom substituted alkyl, heteroatom substituted alkenyl, heteroatom substituted alkynyl, aryl, arylalkyl, arylalkenyl or arylalkynyl.

Abstract: A method of detecting via a solid-phase assay the amount of biological activity, identity and/or the quantity of a biologically active substance is disclosed. The method utilizes the biological activity of the substance itself to provide the method of detection. The method provides competitive and noncompetitive assays.

Abstract: Oxidative stress in living organisms is determined in a photochemiluminescence measuring system after separation of low-molecular weight antioxidants from protein-containing test samples that were withdrawn from these organisms using an assay kit that contains a gel chromatographic column, a photosensitizer solution, a carbonate buffer solution, and a phosphate buffer solution.

Abstract: A substrate having an extended shelf life for chemiluminescent detection and assaying is useful with enzyme-based probes as well as for immunoassays, DNA or RNA or protein detection and blotting. The substrate is a mixture of (a) a chemiluminescent compound, (b) an oxidant, (c) a stabilizer and (d) a buffer. The substrate may further include a chemiluminescent enhancer and a solubilizer. The substrate may be in an organic solvent or may be aqueous based.

Abstract: Disclosed is a method for determining a white blood cell count of a whole blood sample, which comprises: mixing a whole blood sample with a surfactant to thereby obtain a mixture; allowing the mixture to stand for a time sufficient to lyze the white blood cells contained in the whole blood sample and release intrinsic myeloperoxidase from the white blood cells; measuring the concentration of the released myeloperoxidase in the mixture; and determining the white blood cell count in the whole blood sample, based on the concentration of the released myeloperoxidase. Also disclosed is a method of the present invention, in which, in addition to a white blood cell count of a whole blood sample, the concentration of C-reactive protein contained in the same whole blood sample is measured.

Abstract: The invention relates to the detection of apoptotic cells using a novel assay that employs ligation of DNA fragments in situ to selectively label apoptotic cells. The assay may be used in combination with known in situ methodologies to simultaneously detect apoptotic cells and specific biomolecules present in the apoptotic cell.

Abstract: A method for enhancing the conversion of a phenol substrate to a product by a peroxidase enzyme comprises the steps of reacting a conjugate comprising a detectably labeled phenol with a peroxidase enzyme in the presence of an enhancing reagent, the enhancing reagent comprising an inorganic salt, an organic compound having the formula wherein when X is B(OH)2, Y is I, or wherein when X is OH, Y is a halogen, or Q-R wherein Q is a linear or branched 1-12 heteroatom alkyl wherein tie heteroatoms are selected from C, N, O, and S, wherein the bonds connecting the heteroatom alkyl chain are single or double, wherein any carbon atom in the heteroatom alkyl chain optionally includes a substituent selected from —OH, —COOH, —NH2, and —SH, and wherein R is selected from —OH, —COOH, —NH2, and —CH3; or mixtures of the inorganic salt and the organic compound.

Abstract: An electrochemical affinity assay system for detection of ligand—ligand receptor binding.

Abstract: The present invention relates to a method for detecting the amount of a target polynucleotide in a sample. A combination is provided in a medium. The combination comprises (i) a sample suspected of containing the target polynucleotide, the target polynucleotide being in single stranded form, (ii) a reference polynucleotide comprising a sequence that is common with a sequence of the target polynucleotide, and (iii) a predetermined amount of an oligonucleotide probe that has a sequence that hybridizes with the sequence that is common. The combination is subjected to conditions for amplifying the target polynucleotide and the reference polynucleotide. The conditions permit formation of substantially non-dissociative complexes of the target polynucleotide and the reference polynucleotide, respectively, with the oligonucleotide probe. Furthermore, the predetermined amount of the oligonucleotide probe is less than the expected amount of the amplified target polynucleotide.

Abstract: A method for treating hair, combining permanent dyeing and straightening of hair, without significantly damaging the hair. According to the method of the present invention the hair is treated by chemically reducing covalent disulfide linkages in the hair, and contacting said hair with at least one oxidoreductase, at least one mediator, and at least one chemical oxidizing agent in an amount equivalent to 0.001-1% hydrogen peroxide calculated by weight of the dyeing formulation.

Abstract: A method for determining the presence of an analyte in a fluid is described along with various components of an apparatus specifically designed to carry out the method. The method involves taking a reflectance reading from one surface of an inert porous matrix impregnated with a reagent that will interact with the analyte to produce a light-absorbing reaction product when the fluid being analyzed is applied to another surface and migrates through the matrix to the surface being read. Reflectance measurements are made at two separate wavelengths in order to eliminate interferences, and a timing circuit is triggered by an initial decrease in reflectance by the wetting of the surface whose reflectance is being measured by the fluid which passes through the inert matrix. The method and apparatus are particularly suitable for the measurement of glucose levels in blood without requiring separation of red blood cells from serum or plasma.