16 results about "Acetylhomocholine" patented technology

The invention discloses an enzyme biosensor for rapid detection of organophosphorus pesticide dimethoate as well as preparation and application of the enzyme biosensor, and relates to the field of electrochemical sensors. The method comprises the following steps: preparing GR / GCE, preparing AuNPs / GR / GCE, and preparing AChE / AuNPs / GR / GCE. Graphene and gold nanoparticles are compounded to form AuNPs / GR / GCE, the electrochemical impedance of the glassy carbon electrode is remarkably reduced, the effective specific surface area of the glassy carbon electrode is increased, the electron transfer rate of the surface of the glassy carbon electrode is increased, rich binding sites are provided for immobilization of enzyme molecules, the stability of enzyme is improved, and degradation or inactivation of the enzyme in the using process is reduced; acetylcholine esterase is used as a recognition probe, AChE / AuNPs / GR / GCE is constructed on AuNPs / GR / GCE in a self-assembly mode, dimethoate can prevent acetyl thiocholine iodide from being hydrolyzed to generate thiocholine, electrochemical response signals are reduced, and the dimethoate has good electrochemical response signals, so that the organophosphorus pesticide dimethoate residue is rapidly, accurately and safely screened on site.

The invention provides a method for rapidly detecting pesticide residues in fruits and vegetables, and belongs to the field of pesticide analysis and detection. The method comprises the following steps: mixing a copper nanoparticle / reduced graphene oxide / chitosan composite material, acetylcholin esterase, choline oxidase and a cross-linking agent glutaraldehyde solution for reaction to obtain a double-enzyme functionalized nano composite material; dispensing a chitosan solution on the surface of a working electrode of a screen-printed electrode, drying to form a film, and covalently modifying the surface of the electrode with the bienzyme functionalized nano composite material through a cross-linking agent glutaraldehyde to obtain a sensing electrode; according to the method, acetylcholine is taken as a substrate, a sensing electrode is used, a chronoamperometry is adopted, inhibition ratios of pesticide standard substance solutions with different known concentrations to electrode response current are detected, and a standard curve between the pesticide concentration and the current inhibition ratio is established; and calculating the pesticide residue concentration in the fruit and vegetable sample to be detected according to the current inhibition ratio of the liquid to be detected and the standard curve. Therefore, the sensitivity and accuracy of pesticide residue detection in fruits and vegetables are improved.

The application provides a kind of gold nanoclusters with polypeptide as ligand and detection method of methyl parathion, the sequence of polypeptide in the gold nanoclusters with polypeptide as ligand is H (S, D) GGFFYCC;The gold nanoclusters with polypeptide as ligand can be used for double signal detection of methyl parathion and detection of methyl parathion in fruits and vegetables.Fluorescence signal detection: gold nanoclusters are added to the test solution, 37 DEG C reaction 30-60 min, detect the fluorescence change at 700 nm.Colorimetric signal: in pH=8.0 buffer solution, different concentrations of methyl parathion, acetylcholinesterase, choline oxidase and acetylcholine are added, 37 DEG C preheating 30-60 min.pH=4.0 buffer solution, gold nanoclusters and TMB are added, 37 DEG C reaction 10 min, 652 nm is measured absorbance.The detection method of the application has high detection sensitivity, strong anti-interference ability, low detection limit, high correlation coefficient of detection curve, good reproducibility in fruit and vegetable detection recovery experiment, and has good practical value in the detection of methyl parathion.

The invention includes a chimeric autoantibody receptor (CAAR) specific for anti-acetylcholine receptor (AChR) B cell receptor (BCR), compositions comprising the CAAR, polynucleotides encoding the CAAR, vectors comprising a polynucleotide encoding the CAAR, and recombinant cells, e.g., T cells comprising the CAAR.

The present application provides a biological product and a method and an application thereof. The biological product is used for the preparation of a therapeutic agent for cancer treatment, and it includes at least one of the recombinant butyrylcholinesterase-albumin (rBChE-albumin) fusion protein and the recombinant butyrylcholinesterase-Fc (rBChE-Fc) fusion protein. The biological product adjusts the balance of acetylcholine and other related metabolic pathways by supplementing at least one of the exogenous rBChE-albumin fusion protein and rBChE-Fc fusion protein to treat cancer. The present invention solves the problem that the extraction and purification of BChE from human serum may not meet the huge demand for cancer treatment. The present application further provides a biological product for neutralizing and degrading nerve agents and organophosphate compounds, hydrolyzing ghrelin, and anti-wrinkle.

The present application relates to a controllable multi-enzyme cascade AChE-HRP-CHO@HNF composite material preparation and plant wearable sensor application, belong to biosensing, enzyme immobilization material and plant in-situ rapid detection field. The composite material is prepared by biomimetic mineralization and layer-by-layer protein crown assembly, takes enzyme-inorganic hybrid nanoflower as carrier, realizes spatial ordered fixation of choline oxidase (CHO), horseradish peroxidase (HRP) and acetylcholinesterase (AChE) in nanostructure, and forms hierarchical distribution multi-enzyme cascade system. The structure can shorten the transmission distance of intermediate product in cascade reaction, improve the cascade catalytic efficiency and pesticide inhibition response sensitivity. Further embedding it in double network hydrogel, a hydrogel sensing disc suitable for plant surface attachment can be prepared, the hydrogel sensing disc has good flexibility, surface adaptability and storage stability, can be used for on-site visual detection of organophosphorus pesticides such as chlorpyrifos, has the characteristics of simple operation and non-invasive detection.

The application discloses a FRET fluorescent probe with dual response to acetylcholinesterase and viscosity, a preparation method and application thereof, wherein the structure of the FRET fluorescent probe is shown in the following formula: The fluorescent probe is based on a fluorescence resonance energy transfer (FRET) mechanism, responds to acetylcholinesterase (AChE) at a short wave of 520 nm, and the detection limit is as low as 0.104 U / L, and responds to viscosity at a long wave of 680 nm, and the wavelength difference is as high as 160 nm, and the dual-channel response has no crosstalk. Cell toxicity test shows that the probe has good biocompatibility, and confocal fluorescence microscopic imaging experiment shows that the probe has good light stability for cells, has the ability of simultaneously monitoring the concentration change of AChE in cells and the viscosity fluctuation of cells, and is suitable for confocal fluorescence imaging for tracing the concentration change of AChE in cells and the viscosity fluctuation.

The application is suitable for the technical field of pesticide detection, and provides a method for double-signal detection of trace residues of organophosphorus pesticides based on gold nanoscale enzyme, polypeptide is used as a protective agent to synthesize gold nanoscale enzyme, the double properties of fluorescence and peroxidase-like activity of the gold nanoscale enzyme are used, and the enzyme inhibitor method is combined to establish a detection method, and the specific steps are as follows: step one, synthesizing gold nanoscale enzyme and storing for standby; step two, preparing sodium alginate hydrogel blocks, and physically adsorbing gold nanoscale enzyme into the sodium alginate hydrogel through soaking and storing for standby; step three, establishing an acetylcholinesterase / choline oxidase cascade reaction to generate H2O2; step four, based on the H2O2 generated in step three, and combining the prepared sodium alginate hydrogel in step two, realizing rapid visual quantitative detection of OPs, the double-signal output detection result of the application has self-correction, can resist interference, improves detection accuracy, and expands the practical application range of detection.

The application provides an organic phosphorus poisoning relief composite enzyme and application thereof, and belongs to the technical field of biological medicine. The organic phosphorus poisoning relief composite enzyme provided by the application comprises polyethylene glycol modified organic phosphorus hydrolase and butyrylcholine esterase, and the mass ratio of the polyethylene glycol modified organic phosphorus hydrolase to the butyrylcholine esterase is (4-8):(2500-10000). The rat acute organic phosphorus poisoning treatment result shows that the organic phosphorus poisoning relief composite enzyme provided by the application fully plays the efficient catalytic hydrolysis capacity of the enzyme as a catalyst, efficiently removes organic phosphorus and acetylcholine by using the organic phosphorus hydrolase and the butyrylcholine esterase respectively, provides a drug combination for treating the cause for the organic phosphorus poisoning injury first aid, and the treatment effect is significantly better than that of the single drug.

The use of deep eutectic solvents in enzyme-based biosensors improves the robustness of the sensors with regard to storage conditions. For example, phosphotriesterase or acetylcholinesterase enzymes can be used to detect organophosphates using electrochemistry.

The invention discloses an acetylcholin esterase and butyrylcholin esterase double-target inhibitor, which has the following chemical structural formula: wherein R is selected from any one of the following structural formulae;

The application provides a preparation method of metal-doped carbon dots, comprising the following steps: drying an aqueous solution of reactants containing ethylenediaminetetraacetate, Cu 2+ and Fe 3+ , high-temperature pyrolysis, acid washing, water washing, methanol purification and filtering, and drying at 60 DEG C to obtain the metal-doped carbon dots. The phenol measurement steps provided by the application are as follows: mixing a sample solution with a buffer solution, a 4-aminoantipyrine solution and a carbon dot solution, incubating to obtain a solution A, and measuring the absorbance of the solution A at 509 nm for quantitative analysis. The measurement steps of dimethoate provided by the application are as follows: mixing a sample solution with a acetylcholinesterase solution, incubating for half an hour, adding a acetylthiocholine chloride solution and incubating for another half an hour, adding a solution A, and incubating for half an hour before fluorescence measurement. The synthesis method provided by the application is simple, the detection method has high sensitivity and small interference, and is suitable for simultaneous detection of phenol and dimethoate in actual samples.

The invention relates to a large Stokes shift cholinesterase near-infrared fluorescent probe and a construction method thereof.Quinoline is used as a core skeleton, and a conjugated system is constructed through a D-pi-A electronic regulation strategy: aryl, alkenyl or amido is introduced to the 4th site / 6th site to regulate the emission wavelength; the construction method comprises the following steps: synthesizing a fluorophore main skeleton through a Wittig reaction / Heck reaction, connecting a recognition unit through a substitution reaction, and introducing the electron-withdrawing group through a Najing condensation reaction. Stokes shift breaks through 100 nm, and excitation light interference can be inhibited. According to the probe, a recognition unit simulating acetylcholine / butyrylcholine'dumbbell-shaped 'conformation is covalently connected to a quinoline nitrogen atom through a benzyl bromide group, the recognition unit comprises an aromatic cavity adaptive to AChE / BChE substrate pocket hydrophobicity and an active site for catalyzing ester bond hydrolysis, an enzymatic reaction triggers intramolecular electron rearrangement and C-N bond breakage, a fluorescence quenching state is relieved, and the quinoline nitrogen atom can be detected. And "off-on" type signal conversion is realized.

The invention relates to a paper-based sensor, a preparation method of the paper-based sensor and application of the paper-based sensor in detection of alternariol, and belongs to the technical field of food detection. The paper-based sensor comprises a first protective layer, an immobilization layer, a substrate layer, a color development layer and a second protective layer, the surfaces of the first protective layer and the second protective layer are coated with hydrophobic materials; central areas of the immobilization layer, the substrate layer and the color development layer are coated with acetylcholin esterase, acetylthiocholine and nano-enzyme respectively to form hydrophilic reaction areas, and non-hydrophilic reaction areas are coated with hydrophobic materials to form hydrophobic isolation areas; the first protection layer, the immobilization layer, the substrate layer, the color development layer and the second protection layer are folded according to a preset sequence, and the hydrophilic reaction areas of the immobilization layer, the substrate layer and the color development layer are mutually overlapped after folding. The paper-based sensor is remarkably improved in the aspects of detection sensitivity, stability and convenience.

The invention provides a ternary complex dual-mode probe and an acetylcholin esterase activity and enzyme inhibitor detection method thereof. The ternary complex dual-mode probe is composed of a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) system, a manganese dioxide nanoflower-CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) activated ssDNA (ssDNA) (MnO2 NFs-ssDNA) and internal reference fluorescence. When a target to be detected is acetylcholin esterase (AChE), hydrolysis of an acetylthiocholine substrate can be catalyzed, a thiocholine product can be generated, MnO2 NFs can be decomposed to release ssDNA, the trans-cleavage activity of Cas12a can be activated, fluorescence report nucleic acid can be cleaved, a fluorescence signal can be generated, and a more accurate ratio fluorescence signal can be output by taking internal reference fluorescence added into a system as an internal standard; the decomposition of the MnO2 NFs reduces the oxidase-like activity of the MnO2 NFs, and the colorimetric signal for catalyzing the color development of 3, 3 ', 5, 5'-tetramethyl benzidine is reduced; therefore, the higher the AChE activity is, the larger the yield of the thiocholine is, the more thorough the decomposition of the MnO2 NFs is, the stronger the fluorescence signal is, and the weaker the colorimetric signal is. The enzyme inhibitor organophosphorus pesticide can inhibit generation of thiocholine, and then an opposite experiment result appears.

This invention discloses a method for detecting acetylcholinesterase based on a cobalt-doped graphitic carbon nitride photoresponsive nanozyme, comprising the following steps: first, preparing a Co / g-C3N4 nanozyme; then, mixing acetylcholinesterase with buffer and acetylthiocholine, incubating, and subsequently adding buffer, Co / g-C3N4, and TMB; irradiating with a xenon lamp, centrifuging, and recording the ultraviolet-visible absorption spectrum of the supernatant. This method uses a photoresponsive nanozyme, eliminating the need for unstable hydrogen peroxide, and is extremely simple to operate. The catalytic colorimetric reaction of the photoresponsive nanozyme can be precisely controlled by switching the light source on and off, resulting in more accurate detection results. It also provides guidance for the preparation of highly catalytically active g-C3N4-based photoresponsive nanozymes.