222 results about "Enzyme binding" patented technology

A reagent is suitable for measuring the concentration of an analyte in a hemoglobin-containing biological fluid, such as whole blood. The reagent comprises a flavin-dependent enzyme that has specificity for the analyte, a flavin cofactor if, and only if, a flavin is not bound to the enzyme, a tetrazolium dye precursor, an electron transfer agent, and a nitrite salt. The reagent causes dye formation that is a measure of the analyte concentration. The nitrite salt suppresses interfering dye formation caused non-enzymatically by the hemoglobin. Preferably, the reagent is used in a dry strip for measuring glucose in whole blood.

The description relates to cereblon E3 ligase binding compounds, including bifunctional compounds comprising the same, which find utility as modulators of targeted ubiquitination, especially inhibitors of a variety of polypeptides and other proteins which are degraded and / or otherwise inhibited by bifunctional compounds according to the present disclosure. In particular, the description provides compounds, which contain on one end a ligand which binds to the cereblon E3 ubiquitin ligase and on the other end a moiety which binds a target protein such that the target protein is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of that protein. Compounds can be synthesized that exhibit a broad range of pharmacological activities consistent with the degradation / inhibition of targeted polypeptides of nearly any type.

The description relates to cereblon E3 ligase binding compounds, including bifunctional compounds comprising the same, which find utility as modulators of targeted ubiquitination, especially inhibitors of a variety of polypeptides and other proteins which are degraded and / or otherwise inhibited by bifunctional compounds according to the present disclosure. In particular, the description provides compounds, which contain on one end a ligand which binds to the cereblon E3 ubiquitin ligase and on the other end a moiety which binds a target protein such that the target protein is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of that protein. Compounds can be synthesized that exhibit a broad range of pharmacological activities consistent with the degradation / inhibition of targeted polypeptides of nearly any type.

A process for the enzymatic hydrolysis of cellulose to produce a hydrolysis product comprising glucose from a pretreated lignocellulosic feedstock and enzymes for use in the process are provided. The process comprises hydrolyzing an aqueous slurry of a pretreated lignocellulosic feedstock with cellulase enzymes, one or more than one β-glucosidase enzyme and a binding agent for binding the β-glucosidase enzyme to fiber solids present in the aqueous slurry. During the hydrolysis, both the cellulase enzyme and β-glucosidase enzyme bind to the fiber solids. The hydrolysis is performed in a solids-retaining hydrolysis reactor so that unhydrolyzed fiber solids and bound enzyme are retained in the reactor longer than the aqueous phase of the slurry.

Methods are provided for detecting methionine aminopeptidase (MAP) activity and for detecting inhibitors of MAP. The methods utilize a peptide comprising an N-terminal methionine which can be cleaved from the peptide by MAP, and a C-terminal detection moiety which is released by a second peptidase only if the N-terminal methionine has been cleaved from the peptide. When the peptide is combined with MAP and the second peptidase, the detection moiety is released, while the addition of a MAP inhibitor will inhibit the release of the detection moiety. Reaction mixes, peptides, and kits which are useful for practicing the methods are also provided.

Embodiments of the invention include sensors comprising high electron mobility transistors (HEMTs) with capture reagents on a gate region of the HEMTs. Example sensors include HEMTs with a thin gold layer on the gate region and bound antibodies; a thin gold layer on the gate region and chelating agents; a non-native gate dielectric on the gate region; and nanorods of a non-native dielectric with an immobilized enzyme on the gate region. Embodiments including antibodies or enzymes can have the antibodies or enzymes bound to the Au-gate via a binding group. Other embodiments of the invention are methods of using the sensors for detecting breast cancer, prostate cancer, kidney injury, glucose, metals or pH where a signal is generated by the HEMT when a solution is contacted with the sensor. The solution can be blood, saliva, urine, breath condensate, or any solution suspected of containing any specific analyte for the sensor.

This invention relates to novel quinoline-based divalent metal ion chelating ligands of Formula I, wherein A or B are independently —CR7R8, or —CH(R9)CH(R10). X is hydrogen, C1-C10 alkyl; —OH, or —NR11R12. R1 to R12 are various substituents selected to optimize the physicochemical and biological properties such as enzyme binding, tissue penetration, lipophilicity, toxicity, bioavailability, and pharmacokinetics of compounds of Formula 13. R1 to R12 may include, but are not limited to hydrogen, alkyl, acyl, hydroxyl, hydroxyalkyl, substituted or unsubstituted aryl, amino, aminoalkyl, alkoxyl, aryloxyl, carboxyl, halogen, alkoxycarbonyl, cyano, and other suitable electron donating or electron withdrawing groups. The compounds of the present invention are useful for inhibiting the activity of viral enzymes responsible for the proliferation of human immunodeficiency virus (HIV).

The invention refers to a kind of detecting reagent box and the manufacturing method, concretely refers to the reagent which is indirect enzyme immune sorption experiment for detecting rabies virus IgG and the manufacturing method. The reagent box compositions are: beforehand enclosed rabies virus antigen enzyme label board, sample diluting solution, HRP-rabies resisting IgG enzyme compound, condensed washer solvent, substrate and stopping liquid. The specificity of the reagent can reach 100%; the sensitivity is 1:640; the accuracy (the variation coefficient) is 6.98%. The reagent uses indirect ELISA to detect the rabies virus IgG antibody.

The invention relates to a riemerella anatipestifer antibody indirect ELISA method detection kit and an application thereof; the kit comprises: a) an antibody detection plate, b) enzyme conjugate working fluid, c) a positive control, d) a negative control, e) a sample diluent, f) 10X concentrated washing liquid, g) a substrate coloured solution A, h) a substrate coloured solution B, and i) a stopping solution; the beneficial and positive effects of the invention are that: the kit is simple in operation; the requirements for apparatuses needed are not high; the kit can be operated by everyone, can meet the requirements with different levels such as epidemic disease monitoring, hygienic epidemic prevention, intensive culture, and individual culture, is suitable for large-scope popularization and application, and has wide market prospects.

The invention provides a method for modifying the surface of a solid material (e.g. a polymeric matrix). The method is versatile and can be used to prepare polymeric matrices having altered, improved, or specifically engineered properties. Additionally, the method can be used to prepare polymeric matrices that have reactive groups that can be used to immobilize upon the matrices a variety of other “ligand” groups, e.g. a bio-selective affinity group, a chromophore, a dye, an amphiphile, a chiral group, a peptide, a protein, an antibody, an amino acid, an ion exchange group, a detectable group, a carbohydrate, a nucleic acid, a catalyst, a substrate for enzyme binding thereto, an enzyme inhibitor or enzyme co-factor for enzyme binding, or the like.

The invention discloses a luteinizing hormone nano-magnetic particle chemiluminescence quantitative immunoassay kit. The kit comprises a luteinizing hormone calibration product, a nano-magnetic particle suspension coupled with streptavidin, a biotin-marked luteinizing hormone antibody, a luteinizing hormone antibody enzyme binding substance, a luteinizing hormone quality control product, a chemiluminescence solution A and a chemiluminescence solution B, a 20-times concentrated washing solution and a reaction tube, wherein the used enzyme is horseradish peroxidase, the purity RZ of the horseradish peroxidase is not less than 3.0, and the activity is not less than 250U / mL. In addition, the invention further discloses a preparation method of the kit. Compared with the existing kit, the kit disclosed by the invention has the advantages of high sensitivity, a wide range of measurable concentrations, long effective period of a reagent, high degree of automation in detection and the like, and is simple to operate.

The present invention pertains to a method for determining the sequence of a polynucleotide, the method relying on the detection of a conformational change in an enzyme that interacts with and processes along the polynucleotide. The detection of a conformational change may be carried out by measuring changes in a fluorophore bound to the enzyme.

A composition comprising mesoporous aggregates of magnetic nanoparticles and free-radical producing enzyme (i.e., enzyme-bound mesoporous aggregates), wherein the mesoporous aggregates of magnetic nanoparticles have mesopores in which the free-radical-producing enzyme is embedded. Methods for synthesizing the enzyme-bound mesoporous aggregates are also described. Processes that use said enzyme-bound mesoporous aggregates for depolymerizing lignin, removing aromatic contaminants from water, and polymerizing monomers polymerizable by a free-radical reaction are also described.

The invention discloses a sheep echinococcosis ELISA (enzyme-linked immunosorbent assay) antibody detection kit. The detection kit comprises a sheep echinococcosis recombinant antigen Eg95 pre-coated ELISA plate, a sample diluent, a positive reference solution, a negative reference solution, an enzyme bonder, a scrubbing solution, a substrate color-developing solution and a stop solution. A preparation method of a sheep echinococcosis recombinant antigen Eg95 comprises the steps as follows: firstly, PCR (polymerase chain reaction) amplification is performed on an Eg95 gene segment, then EcoRI and XhoI are used for performing enzyme digestion on a PCR product and a prokaryotic expression vector pET-32a simultaneously, a target segment and the vector after gel recovery are connected to construct a recombinant plasmid pET-EG95, and the recombinant plasmid is transformed into Escherichia coli, induced by IPTG (isopropyl-beta-d-thiogalactoside) for expression and purified by Ni-NTA. According to the kit, recombinant protein Eg95 is used as the antigen to detect a sheep echinococcosis antibody, and the antibody detection kit is high in specificity, high in sensitivity and good in stability.

Methods of repressing expression of a recombinant gene in a cell are provided. The methods include the steps of introducing a transposase DNA binding motif into or adjacent to the gene, and introducing into the cell a transposase that is capable of binding to the transposase DNA binding motif. Methods of producing a population of cells of an organism that vary in their expression of a gene are also provided. The methods involve transfecting the cells with a first polynucleotide sequence encoding the target gene operably linked to a promoter such that the target gene is expressed in the cells, wherein the vector has at least one transposase DNA binding motif within or adjacent to the target gene; and transfecting some of the cells with a second polynucleotide encoding the transposable element operably linked to a second promoter such that the transposable element is expressed in the cells. In these methods, the target gene in the cells transfected with the second polynucleotide exhibits reduced expression when compared to the target gene in the cells that are not transfected with the second polynucleotide. Kits having a first polynucleotide with a gene and a DNA transposase binding motif in or adjacent to the gene, and a second polynucleotide comprising a gene encoding a transposase that is capable of binding to the transposase DNA binding motif are also provided.

The invention relates to a kit for quickly detecting a swine fever antibody. In the method, a pair of specific primers is designed, a relatively conservative gene sequence is cloned, an antigen aiming at a swine fever E2 protein is expressed through a pronucleus expression technology, and, on the basis, the kit containing an enzyme linked plate coated with a high-purity and high activity swine fever virus specificity antigen, an enzyme conjugate of rabbit-anti-swine monoclonal antibody containing an HRP (Horseradish Peroxidase) marker, a TMB (Tetramethylbenzidine) color developing liquid and the like is prepared. The kit can quickly detect the swine fever antibody in blood serum or blood plasma and has strong specificity and high sensitivity.

The invention discloses a Senecavirus ELISA antibody detection kit and a preparation method and application. The Senecavirus ELISA antibody detection kit comprises a Senecavirus VP1 protein-precoatedELISA plate, a blocking solution, a diluted sample solution, an enzyme conjugate, a concentrated washing solution, an enzyme substrate solution and a stopping solution. Senecavirus VP1 protein is usedas a coating antigen for the first time, a kit capable of detecting Senecavirus is established, and the antibody level of the Senecavirus in serum can be rapidly detected; the detection kit is high in sensitivity, good in specificity, good in repeatability and stable in result; the detection kit can be applied to monitoring of the antibody level of the Senecavirus so as to understand the status of the Senecavirus antibody in a whole pig herd; in addition, the detection kit uses an Escherichia coli expression system, and has the advantages of economy and cheapness.

The present invention provides compositions and methods for assessing profiles of catalytically active enzymes in compositions containing a plurality of proteins. In preferred embodiments, the enzyme is a hydrolase, most preferably a cysteine protease. The methods described herein use activity based probes (“ABPs”) that have an affinity moiety for directing the binding of the ABP to one or more catalytically active target enzymes, a reactive group for forming a covalent bond at an active site of the target enzyme(s), and a TAG (e.g., a detectable label, preferably a fluorophore). One or more ABPs may be combined with a protein-containing sample under conditions for binding and reaction of the ABP(s) with target enzyme(s) that are present in the sample. The resulting products may then be used to assess the active enzyme profile of the sample, and can be correlated to the presence, amount, or activity of one or more target enzyme(s) present in the original complex protein mixture.

The present disclosure relates to bifunctional compounds, which find utility as modulators of estrogen receptor (target protein). In particular, the present disclosure is directed to bifunctional compounds, which contain on one end a cereblon, Von Hippel-Lindau ligase-binding moiety, Inhibitors of Apotosis Proteins, or mouse double-minute homolog 2 ligand, which binds to the respective E3 ubiquitin ligase, and on the other end a moiety which binds the target protein, such that the target protein is placed in proximity to the ubiquitin ligase to effect degradation (and inhibition) of target protein. The present disclosure exhibits a broad range of pharmacological activities associated with degradation / inhibition of target protein. Diseases or disorders that result from aggregation or accumulation of the target protein are treated or prevented with compounds and compositions of the present disclosure.

The disclosed dual-sandwich enzyme-linked immunologic diagnosis agent box for hepatitis-B core antibody comprises: a micro-porous reaction plate composed by the hepatitis-B core antigen recombinant with purity more than 90% and 5*104~5*105u / mg titer, and the enzyme compound composed by the hepatitis-B core antigen recombinant marked by HRP. This invention has high specificity and sensitivity and convenient to qualitative or semi-qualitative detection.

The invention discloses an identification method and an application of an insulin mass spectrum peptide graph. According to the method, a V8 enzyme is adopted, and the V8 enzyme is combined with lysylendopeptidase, specific hydrolysis of insulin is carried out, and a peptide graph containing insulin characteristic fragments is established through a liquid chromatograph-mass spectrometer. The method is characterized in that a buffer solution is used for providing an enzymolysis environment with the pH value of 7.5-9.5, wherein a substrate and the enzyme are reacted overnight according to the mass ratio of insulin to V8 enzyme to lysyl endopeptidase of (100-1000) to (100-500) to 1, the reaction liquid is diluted for 10-30 times, a liquid phase separation mass spectrometry detector is adopted for detection, and a characteristic peptide graphs of a variety of insulin are established. By adopting the mass spectrum peptide graph, various different kinds of insulin can be accurately determined, and the method is accurate and reliable.

The mid-pregnancy complex screening agent box comprises the AFP standard, AFP single crone anti body wrapped plate bar, AFP quality control, AFP enzyme conjugates, beta -HCG standard, beta -HCG single crone antibody wrapped plate bar, beta -HCG quality control, beta -HCG enzyme conjugates, sample dilute solution, dense cleansing solution, light emitting bottom solution, plate seal film, plate frame, inspecting the AFP or beta -HCG of the pregnant woman, using special data analysis treatment software, with accuracy, reliability, free from trauma, with wide promotion value.

The invention discloses a method for fast screening a topoisomerase I inhibitor from a natural product, and relates to the analysis field of natural products. The method comprises the following steps of: (1) preparation of an enzyme binding reaction buffer solution; (2) preparation of a to be detected sample; (3) preparation of a standard solution; (4) binding reaction of the to-be-detected sample and normal and inactive topoisomerase I; (5) chromatography-mass spectrometry detection analysis of reaction sample; and (6) enrichment ratio of the topoisomerase I to the inhibitor. The ultrafiltration membrane technology and the chromatography-mass spectrometry united technology united method are applied to the screening of the topoisomerase I inhibitor, the sample analysis speed is fast and the specificity is strong so as to conveniently recognize a medicine ligand combined with a biological target molecule; meanwhile, the sample analysis dosage is less, and the spectrogram information quantity is large so as to realize the fast screening of the lead compound, the method has great advantage on the aspect of researching mutual effect of medicine small molecule ligand and biological macromolecule receptor; the method is suitable for analyzing in-vitro enrichment ratio of the natural product extract or monomer compound to the topoisomerase I inhibitor.

The invention provides a carbohydrate antigen-125 quantitative determination kit and a detection method thereof. According to the scheme, a principle of a double-antibody sandwich enzyme immunoassay chemiluminescence method is adopted to prepare a carbohydrate antigen-125 quantitative determination kit. The kit comprises a CA125 (carbohydrate antigen-125) antibody-coated micro-plate / strip, six calibration material bottles, two quality control bottle, an enzyme conjugate, a washing concentrate bottle, a luminous substrate A1 bottle and a luminous substrate B1 bottle. The kit is applied to large-scale detection and manual detection one by one or group by group and has the beneficial effects of high sensitivity, high accuracy and high reproducibility.