1114 results about "Horseradish peroxidase" patented technology

A small diameter flexible electrode designed for subcutaneous in vivo amperometric monitoring of glucose is described. The electrode is designed to allow “one-point” in vivo calibration, i.e., to have zero output current at zero glucose concentration, even in the presence of other electroreactive species of serum or blood. The electrode is preferably three or four-layered, with the layers serially deposited within a recess upon the tip of a polyamide insulated gold wire. A first glucose concentration-to-current transducing layer is overcoated with an electrically insulating and glucose flux limiting layer (second layer) on which, optionally, an immobilized interference-eliminating horseradish peroxidase based film is deposited (third layer). An outer (fourth) layer is biocompatible.

An electrochemical method for determining immunity of tumor marker includes forming microreaction cell, placing reference and counter electrode above it and fixing antigen molecular functioning film at pool bottom, connecting enzyme labelled antibody to pool bottom at time of only proper quantity of horseradish peroxidase to label tumor marker antibody, obtaining cataltyic current on working electrode by contacting free immune matter with electronic media and having positive ratio of the current to antigen. The microvolume immune determining chip is also disclosed.

A small diameter flexible electrode designed for subcutaneous in vivo amperometric monitoring of glucose is described. The electrode is designed to allow “one-point” in vivo calibration, i.e., to have zero output current at zero glucose concentration, even in the presence of other electroreactive species of serum or blood. The electrode is preferably three or four-layered, with the layers serially deposited within a recess upon the tip of a polyamide insulated gold wire. A first glucose concentration-to-current transducing layer is overcoated with an electrically insulating and glucose flux limiting layer (second layer) on which, optionally, an immobilized interference-eliminating horseradish peroxidase based film is deposited (third layer). An outer (fourth) layer is biocompatible.

A conductive polymer is formed enzymatically in the presence of a polynucleotide template. The method includes combining at least one redox monomer with a polynucleotide template and a redox enzyme, such as horseradish peroxidase, to form a reaction mixture. The monomer aligns along the template before or during the polymerization. Therefore, the polynucleotide template thereby affects the molecular weight and conformation of the conductive polymer. When the conductive polymer is complexed to a polynucleotide duplex, the conformation of the polynucleotide duplex can be modulated by changing the oxidation state of the conductive polymer.

The invention relates to a high-sensitivity trace of protein measuring method based on bead and nm metal probe, comprising: labelling the bead using monoclonal antibody of protein to measure; labelling the polyclonal antibody of protein to measure on nm metal probe and at the same time labelling a DNA probe with biotin label on the nm metal probe; labelling horseradish peroxidase on the DNA probe of nm metal probe by biotin-streptavidin reaction to form nm metal probe; mixing the labelled bead and nm metal probe and protein sample and incubating for a period of time at 37 degree; and then washing the nm metal probes which do not react; developing using TMB development system, therefore the protein can be quantificationally detected. The detection time can be reduced to 1-1.5 hours and the sensitivity can be pg / ml.

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.

The invention relates to a preparation method of an electrochemistry immunosensor for detecting alpha fetoprotein, in particular to a gold-composite nanometer mesoporous SiO2 composite-based preparation method of the electrochemistry immunosensor for determining the alpha fetoprotein. The immunosensor is employed to provide a method for determining the alpha fetoprotein. The preparation method ischaracterized by comprising the steps of: firstly synthetizing a nanometer mesoporous SiO2-nano-gold composite, fixing horseradish peroxidase and a secondary antibody on the composite to form a complex; fixing a primary antibody by using a thionine-graphene complex, thereby preparing the electrochemistry immunosensor for determining the alpha fetoprotein, wherein the immunosensor is successfully used for the determination of the alpha fetoprotein. The electrochemistry immunosensor disclosed by the invention has the advantages of high accuracy, high sensitivity, stability and repeatability, rapidness and convenience for immunoassay determination, can be used for clinical analysis and provides important basis for early diagnosis of cancers.

The invention discloses a method for quantitative detection of biological molecules capable of being metabolized to generate H2O2 in serum by means of a ratiometric fluorescent probe. The method includes firstly preparing copper-nitrogen codoped carbon dots Cu-CDs; then reacting corresponding oxidase and the biological molecules capable of being metabolized to generate H2O2 to generate H2O2; catalyzing the H2O2 and a substrate that is o-phenylenediamine with horseradish peroxidase to generate an oxidation product DAP having yellow fluorescence; then adding the Cu-CDs into the DAP; allowing the DAP and the Cu-CDs to form ratiometric fluorescence, with the ratio I<572> / I<460> of fluorescence intensities of the DAP and the Cu-CDs being in a linear relationship with the concentration of a substance to be detected; and performing quantitative assay of the biological molecules in the serum according to the ratio of fluorescence intensities. The method is high in sensitivity and simple and convenient in detection, and has high selectivity and high affinity of immunoreactions.

The present invention discloses a chloramphenicol chemiluminescence enzyme-linked immunodetection kit, which comprises a kit body, an enzyme label plate placed inside the kit body, and reagents placed inside the kit body, and is characterized in that every hole of the enzyme label plate is coated with coating antigen, the coating antigen is a chloramphenicol and carrier protein conjugate, and the reagents comprise horseradish peroxidase-labeled chloramphenicol monoclonal antibody, a series of chloramphenicol standard solutions, a concentrated phosphate buffer, a concentrated washing solution and a chemiluminescence solution. The chloramphenicol chemiluminescence enzyme-linked immunodetection kit has characteristics of high sensitivity, simple and rapid detection, and high accuracy, provides a substantially reduced operation time compared to the conventional colorimetric ELISA method, and can be used for detection of chloramphenicol residues in animal tissues (pork, chicken, pork liver and chicken liver), aquatic products (fish and shrimp) and milk.

The invention discloses a kit for detecting pig pseudorabies virus antibodies and a block enzyme-linked immuno sorbent assay (ELISA) method. The kit for detecting pig pseudorabies virus antibodies comprises pig pseudorabies virus monoclonal antibodies which are labelled by horseradish peroxidase, wherein the pig pseudorabies virus monoclonal antibodies are monoclonal antibodies obtained by pig pseudorabies viruses as immunogens and the pig pseudorabies viruses are pseudorabies virus strain Ea. The kit for detecting pig pseudorabies virus antibodies also comprises an enzyme label plate, a sample diluent, negative and positive contrasts, a coloured solution, a washing solution, and a stopping solution. The block ELISA method comprises the following steps of 1, taking out a detection plate pre-coated with virus antigens from the kit for detecting pig pseudorabies virus antibodies, adding diluted blood serum needing to be detected into the detection plate pre-coated with the virus antigens, and simultaneously, setting negative and positive contrast apertures, 2, shaking up the diluted blood serum in the negative and the positive contrast apertures, shaking off a solution in the negative and the positive contrast apertures, and washing the detection plate by the washing solution, and 3, adding the pig pseudorabies virus monoclonal antibodies labelled by horseradish peroxidase into the negative and the positive contrast apertures, washing, adding the colored solution into the negative and the positive contrast apertures to carry out room-temperature coloration in the dark, adding the stopping solution into the negative and the positive contrast apertures, and determining OD630nm values of the negative and the positive contrast apertures by an ELISA apparatus. The block ELISA method has the advantages of good singularity, high sensitivity, short detection time, and high accuracy because of utilization of an S / N ratio method in result determination.

A method of crosslinking an oxidative polymer having oxidatively crosslinkable functional groups and a two-pack composition used therein. The oxidatively crosslinkable functional groups on the oxidative polymer are crosslinked by contacting the oxidative polymer with a catalytic amount of an oxidizing enzyme, such as horseradish peroxidase. The present invention is further directed to a two-pack coating composition which includes a polymeric component and a catalytic component, which are mixed together prior to use.

The invention relates to a method for detecting thrombin with an electrochemical aptamer sensor based on aptamer-gold nanoparticle-horseradish peroxidase signal amplification, belonging to the technical field of analytical chemistry, characterized in that: a thrombin aptamer I is fixed on the gold coated magnetic nanoparticles as a capture probe, an aptamer II is double-labeling by gold nanoparticle and horseradish peroxidase to form an aptamer-gold nanoparticle-horseradish peroxidase composite as a detection probe; the capture probe and the detection probe are combined with thrombin to form a [gold coated magnetic nanoparticle-aptamer I] / thrombin / [ aptamer II-gold nanoparticle-horseradish peroxidase] sandwich structure; and the sandwich structure is fixed on the surface of the electrode, and thrombin is detected through enzyme catalytic reaction. The method has the advantages of simple operation, rapid response, high sensitivity, and strong specificity. The detection limit of thrombin is 30 fmol.L<-1>, and the linear range is 0.1-60 pmol.L<-1>.

The invention relates to a nanotechnology-based trace protein detection method, which combines enzyme-linked immunosorbent assay technology, tyramine signal amplification technology and the aggregation phenomenon of gold nanoparticles modified by different biological molecules, so an experimental method used for detecting trace proteins such as prostate specific antigen (PSA) and the like is established. The method comprises the following steps of: fixing an antibody aiming at the protein to be detected (such as the PSA) on the surface of a substrate; incubating another antibody with horseradish peroxidase (HRP) activity of the protein to be detected (such as the PSA) after capturing the protein to be detected in a sample, wherein the HRP catalyzes biotin-tyramide to generate biotin deposition under certain conditions; further amplifying a signal by using the aggregation phenomenon of the gold nanoparticles modified by biotin-labeled DNA and the gold nanoparticles modified by streptavidin; performing silver staining; and performing data analysis on an experimental result by using software. The method has the advantages of extremely low detection limit, wider detection range, capacity of detecting the antigen in a rabbit serum with complex compositions, and important application prospect.

The invention relates to the high-stable increased chemical illuminant base material, which is composed of the solutions A and B. the solution A contains luminal, reinforcing agent phenol and coordinate reinforcing agent; the solution B contains urea peroxide and H2O2. The illuminant signal of the invention is stable and is suitable for the automatic or hand analysis, and the testing results are precise. The 2-8 Deg. C. storing stability can last for two years.

The invention relates to technical field of immunodetection, particularly relates to a kit for joint detection of ovarian cancer tumor markers HE4 and CA125 as well as a preparation method and application thereof. According to the kit provided by the invention, silanization treatment is performed on horse radish peroxidase, so that the possibility that the horse radish peroxidase becomes a phosphoric acid receptor in a process of catalyzing a chemiluminescent substrate to emit light by a alkaline phosphatase is blocked, the problem that the existence of horse radish peroxidase disturbs the cataluminescence of the alkaline phosphatase is solved, so that the size of the light intensity caused by the alkaline phosphatase on magnetic particles is detected by chemiluminescent substrate liquid first, the size of chromogenic absorbancy caused by the horse radish peroxidase is detected by chromogenic substrate liquid and the simultaneous detection on HE4 and CA125 of a sample is realized. Thekit provided by the invention has the advantages that the detection process is simple, the reaction time is short, the reagent dose is less, the cost is reduced, the sensitivity is high, the repeatability and stability of the reagent are good.

The invention discloses a triiodothyronine quantitative detection kit which comprises magnetic particle suspension coated with diiodothyronine-gelatin, a triiodothyronine series calibrator, a triiodothyronine antibody labelled by horseradish peroxidase, a dissociation agent, a chemiluminescent substrate A, a chemiluminescent substrate B and wash concentrate. The invention further discloses a preparation method of the kit. The invention has the advantage that a chemical structural analogue of hormone is coated by a method for coating an antigen analogue with a labelled antibody, and the analogue and the detected hormone have identical immune correlation, thus realizing specific binding of the analogue and the antibody of the hormone; but the binding capability of the analogue with thyroid-binding protein (THBP) is significantly lowered, thus reducing influence of the binding protein on a measuring system to a large extent. The kit has greatly improved sensitivity and precision, greatlyenlarged detection range, greatly shortened reaction time, enhanced product properties and lowered product cost.

The invention provides a method for preparing a temperature-responsive adhesive injectable hydrogel. The method comprises the following steps: synthesizing a temperature-responsive polymer with different dopamine contents from dopamine methacrylamide used as an adhesive monomer and ethyl 2-(2-methoxyethoxy)methacrylate and oligo(ethylene glycol)methyl ether methacrylate as temperature-sensitive monomers, and carrying out enzyme catalytic crosslinking on the polymer and a dopamine group under the catalysis of horseradish peroxidase and hydrogen peroxide to prepare the temperature-responsive adhesive hydrogel. The hydrogel prepared in the invention has the characteristics of fastness in gel formation, realization of in-situ injection molding, realization of regulating the gelation time and the gel strength by regulating the concentrations of the polymer, horseradish peroxidase and hydrogen peroxide, and convenience in use; and the adhesion strength of the hydrogel is enhanced with the rising of the temperature, so the temperature-responsive characteristic makes the obtained hydrogel applied to fields of tissue adhesives, wound dressings, cell culture media and drug controlled releasecarriers.

The invention is directed to transfection reagents for the delivery of nucleic acids into neural cells, compositions including the reagents, methods of preparation of such reagents, methods of transfecting cells with such reagents, and uses thereof. In preferred embodiments the reagents comprise horseradish peroxidase and / or a polycarboxylic acid such as poly(acrylic acid) or poly(methacrylic acid).

The invention relates to a preparation method of a chitosan modified graphite oxide functional leather tanning agent. Currently, the common leather tanning agents are chroming tanning agent, aluminum tanning agent, vegetable tanning agent, aldehyde tanning agent and synthetic tanning agent and the like, but the leather tanning agents can generate pollution problems hard to treat. The preparation method provided by the invention has the following steps of: mixing crystalline flake graphite powder, sodium nitrate and concentrated sulfuric acid and stirring uniformly, slowly adding potassium permanganate for oxygenation, and preparing a suspending liquid through washing, pH adjustment and ultrasonic treatment; and degrading chitosan with high-temperature alpha-amylase so as to obtain a degraded chitosan solution, modifying graphite oxide with the degraded chitosan under a horseradish peroxidase-hydrogen peroxide catalysis system, thus obtaining the chitosan modified graphite oxide functional leather tanning agent. The preparation method provided by the invention has the advantages of unique synthetizing preparation process and good tanning property, is suitable for main tanning and retanning; and the leather has excellent intensity and flexibility after main tanning.

The invention relates to a method for detecting 1,5-dehydration glucitol in blood and a kit, and provides a new method for detecting the content of the 1,5-dehydration glucitol in the blood, which comprises the following steps of: catalyzing the 1,5-dehydration glucitol by using pyranose oxidase to generate 1,5-dehydration fructose and H2O2; generating quinine compounds by using 4-aminoantipyrine(4-AAP), 3-hydroxy-2,4,6-trihydroxybenzoic acid (HTIB) and H2O2 under the catalytic action of horse radish peroxidase; and determining the level of the 1,5-dehydration glucitol in the blood by colorimetric analysis. The invention also provides the kit for the method. The method and the kit are easy and convenient to operate safely, long in stable time and high in interference resistance, specificity and sensitivity, the interference of multiple glucoses in blood samples can be eliminated quickly, and the reliable foundation can be provided for the diagnosis and treatment of diabetes.

The invention provides enzyme-catalyzed disulfide bond-crosslinked natural polymer hydrogel and a preparation method thereof. The hydrogel comprises a three-dimensional net structure composed of crosslink bonds of cystamine or N,N-diacetyl-L-cystine; under the conditions that horseradish peroxidase is used as a catalyst and a phenol compound is used as an enzyme substrate, sulfydryl-grafted natural polymers are rapidly oxidized to form disulfide bonds, so that the hydrogel is obtained. According to the invention, raw materials are safe in source and environment-friendly, the product has excellent biocompatibility; reaction conditions are mild, and the operation is simple; the system is not required to be additionally added with hydrogen peroxide, and physiological activity of a loaded drug or cells is completely guaranteed.

The invention discloses a pig mycoplasma pneumonia recombination antigen ELISA detection Kit. The Kit is provided with an antibody detection plate, enzyme conjugate treatment fluid, a positive control, a negative control, sample diluent, 10x condensed cleaning solution, developing solution A, developing solution B and termination solution. The detection plate of the Kit is a detachable 96-pore enzyme label plate enveloped by the mutational pig mycoplasma pneumonia membrane protein P46 gene protein antigen, the enzyme conjugate treatment fluid is a rabbit anti-pig antibody labeled by horse radish peroxidase, the positive control serum is taken from a pig which is detected positive through indirect hemagglutination and the ELISA Kit of IDEXX and has obvious pig mycoplasma pneumonia lesions in the lungs after anatomy, and the negative control serum is taken from a pig which is detected negative through indirect hemagglutination and the ELISA Kit of IDEXX and has no pig mycoplasma pneumonia lesions in the lungs after anatomy. The pig mycoplasma pneumonia recombination antigen ELISA detection Kit has the advantages of strong specificity, high sensitivity, simple operation, easy large-scale generation and application, and broad market prospect.

The invention discloses a C-peptide micropore plate type magnetic granule chemoluminescence immunoassay measuring kit and a preparation method thereof. The kit comprises: (1) a C-peptide calibration sample, (2) magnetic granules coated by a C-peptide monoclonal antibody, (3) another monoclonal antibody or a polyclonal antibody marked by horse radish peroxidase (HRP), (4) a micropore plate, (5) washing solution and (6) chemoluminescence substrate solution on which the enzyme acts. The preparation method for preparing the kit further comprises the steps of preparing the calibration sample, preparing the magnetic granules coated by the C-peptide antibody, marking the C-peptide antibody with the horse radish peroxidase (HRP), preparing the washing solution and preparing the chemoluminescence substrate solution. The kit is simple, convenient, fast, sensitive, stable and the like.

The invention discloses enzyme-based magnetic nanoparticles, a preparation method, an application and a detection method for glucose. The preparation method comprises the following steps of 1) mixing Fe3O4 magnetic nanoparticles and dopamine with water according to a mass ratio of Fe3O4 magnetic nanoparticles to dopamine being (1:1)-(1:4) uniformly, adding the above mixture in an alkali buffering system, and stirring for 8-48 hours to obtain Fe3O4 coated by polydopamine; 2) mixing a horseradish peroxidase solution, a glucose oxidase solution and Fe3O4 coated by polydopamine with dopamine uniformly to obtain a solution B, and reacting for 5-25 hours. In the solution B, the concentrations of horseradish peroxidase, glucose oxidase and Fe3O4 coated by polydopamine and dopamine are 0.025-0.2 mg / mL, 0.025-0.2 mg / mL, 0.5-25 mg / mL and 1-5 mg / mL respectively. The enzyme-based magnetic nanoparticles can detect glucose and can be used repeatedly, and waste can be treated by combustion or high temperature, so that zero-pollution can be achieved.

The invention relates to an enzyme-linked immunosorbent assay (ELISA) kit for duck hepatitis virus type-I serum antibody and relates to a test method and application of the kit. The kit comprises an enzyme label plate coated by the recombinant VP1 (virus protein) protein, a rabbit anti-duck IgY antibody marked by horseradish peroxidase, a TMB substrate colour reagent, a positive serum, a negative serum and a kit specification. In the invention, by adopting the polymerase chain reaction, the VP1 genes are amplified from the DHV-1genome and the VP1 gene-containing recombinant expression plasmid pET32a-VP1 is constructed; the plasmid is transferred to host cells BL21 (DE3), and the in-vitro expression VP1 protein is purified by a nickel column and then used as the antigen; the enzyme-linked immunosorbent assay kit is established; the positive serum is the standard positive serum of duck hepatitis virus type-I and the negative control is the standard negative serum of duck. The test kit has the advantages of strong specificity, high sensitivity, simple operation, easy large-scale popularization and application, very important application value in diagnosis of duck hepatitis virus type-I, survey of epidemiology and immunization survey and the like.

This invention relates to a kind of enzyme test paper capable of testing hydrogen peroxides in a sample quickly prepared by an enzymatic chromogenic reaction, which utilizes the adsorption action with anion carriers to fix the horseradish peroxide enzymes with positive charges and a developer on a carrier at the same time, which can test the existence of hydrogen peroxides with the concentration to 0.5mumol / L in the sample and the response time of the test paper is 1 minute.