97 results about "Phosphorylated proteins" patented technology

The invention belongs to the field of functional proteomics research, and particularly relates to a combined analysis method for analyzing interaction between a small molecule substance and a targeted protein, which is used for analyzing the small molecule substance and the protein by adopting LiP-MS (Limited Protein-Mass Spectrometry) technology and phosphorylated proteome combined analysis. According to the method, the optimized LiP-MS technology is adopted, the target protein can be directly recognized under the complex proteome background without enriching specific protein, and the problem that low-abundance protein is difficult to detect is solved; phosphorylated proteome abundance transformation protein is analyzed by combining high-resolution mass spectrometry, so that the recognition capability on a drug target and a shear variant thereof is enhanced, and the detection sensitivity is improved.

The invention discloses a magnetic particle chemiluminiscence detection kit for human non-phosphorylated Tau217 protein and a preparation method of the magnetic particle chemiluminiscence detection kit. The kit comprises a micro magnetic particle coupled with a human non-phosphorylated Tau217 protein capture antibody, an acridinium ester labeled human non-phosphorylated Tau217 protein detection antibody, a human non-phosphorylated Tau217 protein calibration product and a quality control product. The direct chemiluminescence method established by the invention has the advantages of high sensitivity, strong specificity, accuracy, rapidness and short detection time, the detection result has higher accuracy and repeatability, and the kit can be used for various luminescence detection instruments.

The invention relates to the technical field of related drugs for nerve injury caused by heavy metal pollution, in particular to application of a compound for inhibiting phosphorylation of ARMC10 in preparation of drugs for antagonizing tin-induced nervous system injury. Exposure of trimethyltin chloride induces nerve cell mitochondrial dysfunction, and the key mechanism is ARMC10 serine 43 site abnormal phosphorylation. Phosphorylated protein causes excessive mitochondrial fission, membrane potential collapse and energy metabolism disorder, resulting in neuron damage. Based on the target spot, a compound for antagonizing tin-induced nervous system injury is obtained through screening, protein phosphorylation can be specifically inhibited, mitochondrial dynamic unbalance and dysfunction induced by trimethyltin chloride are effectively reversed, and neurotoxicity is relieved. According to the technical scheme, the technical problem that in the prior art, no medicine for effectively antagonizing tin-induced nervous system injury exists can be solved, and the compound has the application potential for treating the nervous system injury and cognitive impairment caused by tin exposure.

The invention belongs to the field of antibodies, and particularly relates to a p-Tau217 phosphorylated protein binding antibody or an antigen binding part thereof and application thereof. The invention provides the amino acid sequences of the heavy chain CDR and the light chain CDR of the antibody specifically bound with the p-Tau217 phosphorylated protein, and the antibody can specifically recognize and bind the p-Tau217 phosphorylated protein and has the advantages of high affinity and good specificity to the protein. Based on the advantages, the antibody can be applied to diagnosis of diseases (such as nervous system degenerative diseases) related to abnormal p-Tau217 phosphorylated protein level, and especially can be applied to clinical diagnosis of Alzheimer's disease.

The invention relates to the technical field of antibodies, in particular to antigen peptides of human PP1 alpha protein and Thr320 site phosphorylated protein of the human PP1 alpha protein, and a preparation method and application of an antibody of the antigen peptides of the human PP1 alpha protein and the Thr320 site phosphorylated protein. The invention provides an antigen peptide of human PP1 alpha protein and Thr320 site phosphorylated protein thereof, the antigen peptide has obvious advantages in the aspects of immunogenicity and the like, animals can be induced to generate high-level antibodies, and the generated antibodies have high specificity, affinity and titer. Based on the antigen peptide, the invention provides an anti-human PP1 alpha protein or a polyclonal antibody with phosphorylated Thr320 site thereof and a preparation method thereof, the polyclonal antibody has the characteristics of strong specificity, high affinity and high titer, and through immunoblotting and immunohistochemical verification, the polyclonal antibody can be used for preparing the anti-human PP1 alpha protein or the anti-human PP1 alpha protein or the anti-human PP1 alpha protein or the anti-human PP1 alpha protein or the anti-human PP1 alpha protein. The probe shows good specificity and sensitivity in detection of phosphorylation modification of human PP1 alpha protein and Thr320 site thereof, and has a good application prospect.

The invention provides an anti-human phosphorylated tau217 rabbit monoclonal antibody as well as a preparation method and application thereof, and belongs to the technical field of biological detection and biological engineering. The rabbit monoclonal antibody for resisting the human phosphorylated tau217 comprises a light chain variable region and a heavy chain variable region, and the monoclonal antibody comprises HZK33-B0010, HZK33-B0011 and HZK33-B0029. The invention develops three rabbit monoclonal antibodies for identifying p-tau217, the rabbit monoclonal antibodies do not have cross reaction with non-phosphorylated tau protein or phosphorylated tau protein at other sites, the rabbit monoclonal antibodies can be combined with p-tau217 with high specificity, and a related body fluid marker detection kit with higher specificity is developed on the basis of the rabbit monoclonal antibodies.

The invention relates to a kit and a method for detecting phosphorylated Tau protein. In particular to a p-Tau217 protein detection kit and a p-Tau217 protein detection method. The kit and the method disclosed by the invention are based on the p-Tau217 antibody and the total-Tau protein antibody, and are combined with a TSA technology, so that the specificity, the sensitivity and the accuracy of a detection result are improved, and background signals are effectively reduced.

The application discloses an acridinium ester labeled complex, which comprises a body, and a labeling group labeled on the body, wherein the body is an antibody, and the labeling group is an acridinium ester group. A preparation method of the acridinium ester labeled complex comprises the following steps: sucking a labeling buffer into a centrifugal tube; sucking an antibody to be labeled into the centrifugal tube and mixing to generate a first mixed solution; adding an acridinium ester solution and mixing, and then performing reaction at 25 DEG C in the dark for 2 hours to generate a second reaction solution; adding a termination buffer and mixing, and then performing reaction at 25 DEG C in the dark for 30 minutes to generate a third reaction solution; performing desalting purification on the third reaction solution to generate a fourth mixed solution; and filtering the fourth mixed solution with a 0.22 mu m filter membrane to generate the acridinium ester labeled complex. According to the preparation method, the labeling efficiency of the acridinium ester and the antibody is higher, the reactivity is stronger, the non-specific adsorption is lower, the sensitivity is higher, the repeatability and stability are better, and the labeling method is simple, rapid and easy to repeat.

The invention provides a phosphorylated protein enrichment method, and relates to the fields of materials, analytical chemistry and life science. The enrichment method of the phosphorylated protein comprises the following steps: dissolving a phosphorylated protein sample in a sample loading solution, mixing with super-hydrophilic metal ion functionalized microspheres, hatching, centrifuging, and discarding supernatant to obtain a super-hydrophilic material enriched with the phosphorylated protein; mixing the material enriched with the phosphorylated protein with an elution solution, incubating, and centrifuging to obtain enriched liquid of the phosphorylated protein; the super-hydrophilic metal ion functionalized microsphere is provided with a super-hydrophilic nano convex shell which is completely coated on the solid core. The enrichment method provided by the invention can overcome the non-specific adsorption problem caused by modification defects of a traditional monomolecular layer modified functional material, and has excellent enrichment selectivity; the method can overcome the limitation of an immunoprecipitation enrichment method antibody, has wide applicability, and is hopeful to occupy an important position in phosphorylated protein separation and enrichment.

The application discloses an antibody for detecting phosphorylated Tau protein pTau217, an immunodetection method and application. The antibody comprises a T217-3D6 antibody; light chain CDR1, CDR2 and CDR3 of the T217-3D6 antibody are sequentially the sequences shown in SEQ ID NO. 1 to 3, and heavy chain CDR1, CDR2 and CDR3 are sequentially the sequences shown in SEQ ID NO. 4 to 6. The antibody is used for immunodetection of the phosphorylated Tau protein pTau217, and has the advantages of simple operation, high sensitivity and strong specificity, and can realize rapid detection of the phosphorylated Tau protein pTau217, and has important significance for evaluating the Tau protein phosphorylation level and Tau protein pTau217 phosphorylation related detection.

An optoelectrochemical sensor, a preparation method thereof, and uses thereof. The sensor uses p-type semiconductor Cu₂O modified on an ITO electrode and can generate a cathodic photocurrent signal. Further, the electrode interface is modified with a capture antibody Ab1 and BSA. When the target phosphorylated tau181 protein is present, through antigen-antibody specific recognition, the probe MZIS-Ab2 is introduced. And due to the energy level matching between MZIS and Cu₂O, the photocurrent electron transfer path is changed, so that the photocurrent polarity is reversed to generate an anodic photocurrent. The developed PEC sensor with target-induced photocurrent polarity reversal has excellent anti-interference ability and low background signal, and can avoid false positive and / or negative signals in the highly sensitive analysis of phosphorylated tau181 protein.

The invention relates to the field of protein detection, in particular to a method for detecting phosphorylated Tau-217 protein in urine of an Alzheimer's disease patient, which comprises the following steps of: 1, collecting fresh urine in the morning by using a urine collecting container, and 2, taking out 100 microliters of urine by using a transfer pipette and adding the urine into a 5.0 ml centrifugal tube; step 3, adding 4.9 ml of a urine diluent to dilute the urine; and step 4, prefabricating tracing fluorescent microspheres and enrichment microspheres in the centrifugal tube. Step 5, repeatedly reversing and uniformly mixing, and reacting at room temperature for 8-12 hours; step 5, placing the reaction tube on a separation test tube rack with a magnet for more than 20 minutes; step 6, irradiating the test tube by using a highlighter with the wavelength of about 340nm in a dark environment to judge a result; and step 7, if a red fluorescent dot appears near the magnet in the test tube, the result is positive, otherwise, the result is negative, and the result can also be judged by measuring a fluorescence value through a luminoscope with the wavelength of 340nm. Finally, the phosphorylated Tau-217 protein is detected, and the operation threshold and the economic cost are reduced.

Provided herein are compositions and methods relating to improved assays for establishing Alzheimer's disease. Further provided herein are compositions and methods comprising improved antibodies for assays including immunoassays.

The application discloses a phosphoric acid modified protein or polypeptide, a preparation method thereof and application thereof in vaccines, and belongs to the field of protein phosphoric acid modification. The preparation method of the phosphoric acid modified protein or polypeptide is as follows: a compound containing an aldehyde group and a phosphate group is subjected to nucleophilic reaction with a protein or polypeptide to form a Schiff base; a reducing agent is used to reduce the Schiff base, and after post-treatment, the phosphoric acid modified protein or polypeptide is obtained. After the phosphoric acid modified protein product prepared by the method is prepared into a vaccine with an aluminum hydroxide adjuvant and mice are immunized, the antibody titer is significantly increased. This is of great significance for increasing the immune effect, reducing the inoculation dose, saving the antigen amount and improving the immune response of some people or in specific conditions.

The invention relates to the field of material analytical chemistry and biochemistry, and provides a continuous batch pretreatment device and method for proteomics, glycoproteomics and phosphorylated proteomics samples. According to the continuous batch pretreatment device for the proteomics, glycoproteomics and phosphorylated proteomics samples, under the assistance of the filler, the whole process of protein denaturation, aggregation capture, enzymolysis and complete glycopeptide and phosphorylated peptide enrichment in biological samples can be achieved. Compared with a free solution method conventionally used in proteomics, the sample pretreatment method based on the device has the advantages that the sample treatment time can be effectively shortened, and continuous analysis of mass proteome, glycoprotein group and phosphorylated proteome of trace protein samples can be realized; the method is expected to be widely applied to the aspects of disease occurrence mechanisms, drug action mechanisms, clinical detection, biomarker discovery and the like.

The invention relates to methods and kits for assessing brain injury, e.g., traumatic brain injury resulting from blast exposure. The invention provides methods of quantifying the amount of phosphorylated tau or total tau in a biological sample. The invention further provides a method of determining the number of blast exposures experienced by a subject. Also provided herein are kits for detecting phosphorylated tau or total tau in a biological sample.

The application discloses an amino-functionalized magnetic mesoporous nanomaterial and a preparation method and application thereof, and the nanomaterial is Fe3O4@nSiO2@TiO2@mSiO2-NH2; from inside to outside, the nanomaterial comprises a magnetite magnetic core, a dense silica layer, a titanium dioxide layer and an amino-functionalized mesoporous silica layer. The magnetic core is synthesized by a solvothermal method, the dense silica layer and the titanium dioxide layer are coated by adopting a sol-gel method in sequence, and finally the amino-functionalized mesoporous silica layer is modified by a co-condensation method. The material can protonate the surface amino group to form a local alkaline microenvironment under an acidic condition, can maintain trypsin activity, realizes synchronous performance of rapid enzymolysis of proteins and in-situ enrichment of phosphorylated peptides, and significantly shortens the enzymolysis time. The application has excellent enrichment selectivity for the phosphorylated peptides, the sensitivity reaches 10 fmol / μL, the enrichment capacity is about 100 mg / g, and is suitable for high-throughput quantitative analysis of phosphorylated proteins in complex biological samples.

The invention belongs to the technical field of medicines, and particularly relates to application of arginine in repairing skeletal muscle injury caused by monensin poisoning. It is found for the first time that arginine can remarkably recover the normal function of skeletal muscle mitochondria and the normal polymerization capacity of microtubules by up-regulating the expression level of mitochondrial oxidative phosphorylation (OXPHOS) protein complex and regulating microtubule polymerization kinetics and reversing monensin-induced skeletal muscle mitochondrial dysfunction and microtubule network disintegration; the method provides a theoretical basis for clinical transformation, and has good practical application value.

This invention belongs to the field of active ingredient extraction and purification technology, specifically a neutral polysaccharide ORP-0 from *Agaricus bisporus* and its applications. The preparation method of this invention includes the following steps: (1) pretreatment; (2) polysaccharide extraction; (3) impurity removal; and (4) separation and purification. This invention obtains a single neutral polysaccharide ORP-0 through defatting pretreatment, ultrasonic-assisted extraction, multiple impurity removal processes, and separation and purification using a DEAE-52 cellulose chromatography column. The method of this invention is simple to operate, has mild reaction conditions, and a complete impurity removal process. The obtained neutral polysaccharide has high purity, with a total sugar content of 92.09%. Furthermore, the obtained neutral polysaccharide ORP-0 enhances the activity of RAW 264.7 macrophages, increases the release of immune factors, enhances the expression of phosphorylated proteins in the MAPK signaling pathway, and activates the body's immune response.

The invention relates to the field of biomedicine, in particular to an oral cancer screening and prognosis kit based on detection of phosphorylated Cofilin protein expression level and application of the oral cancer screening and prognosis kit. The kit contains xylene, alcohol, a citrate buffer solution, hydrogen peroxide, a confining liquid, a primary antibody, a secondary antibody, a DAB developing solution, hematoxylin and neutral gum. According to the kit, the malignant process of the oral squamous cell carcinoma is judged by detecting the expression level of the phosphorylated Cofilin protein in the oral mucosa tissue of a human body, the kit is used for auxiliary diagnosis of clinical oral squamous cell carcinoma, an effective theoretical basis is provided for taking targeted blocking measures or decisions for patients, and the clinical application and transformation prospects are good.

The invention relates to the technical field of protein detection, in particular to a phosphorylated Tau 181 protein detection card, application of the phosphorylated Tau 181 protein detection card and a detection kit comprising the phosphorylated Tau 181 protein detection card. The phosphorylated Tau 181 protein detection card comprises a card shell and a detection test strip arranged in the card shell, the detection test strip comprises a bottom lining, and a sample pad, a coating film and water absorption paper which are sequentially overlapped and adhered to the bottom lining along the length direction of the bottom lining; a microsphere line is sprayed on the sample pad; and the microsphere line is a phosphorylated Tau 181 monoclonal antibody and a goat anti-rabbit IgG (Immunoglobulin G) antibody which are marked by bimodal nano probe microspheres. The detection kit comprises a phosphorylated Tau 181 protein detection card and an ID card containing a calibration curve. The phosphorylated Tau 181 protein detection kit provided by the invention has the characteristics of wide detection range and high sensitivity, and can quickly and accurately detect the content of phosphorylated Tau 181 in human blood.

Provided herein are compounds, methods and compositions for determining whether a patient has a neurological disease or disorder is provided, comprising detecting the presence of a target protein, or an accumulated mass thereof, for example, amyloid beta protein or phosphorylated tau protein, or an accumulated mass thereof, in a tissue or a sample of the patient. The detecting may comprise contacting the target protein with a compound described herein.

The application provides an application of a modified form of GMFB, GMFBS53 phosphorylated protein, as a biomarker in preparation of a MASH early diagnosis reagent. The application also provides a kit for quantitatively or semi-quantitatively detecting a protein, which contains an antibody capable of specifically combining with the GMFBS53 phosphorylated protein. The application can more efficiently and accurately perform early diagnosis on a subject suffering from MASH by detecting the expression level of the GMFBS53 phosphorylated protein, and provides an important basis for early prevention and treatment of potential liver cancer patients. The kit can be applied to early diagnosis of MASH.

According to the phosphorylated protein-polypeptide conjugate and the preparation method thereof, polypeptide in the phosphorylated protein-polypeptide conjugate has threonine (Thr) phosphorylation modification, phosphorylation coupling sites can be accurately positioned, the phosphorylated protein-polypeptide conjugate has high thermal stability, and the thermal stability of the phosphorylated protein-polypeptide conjugate is remarkably higher than that of wild type Tau protein in long-term storage. The method can be used as a quality control product and has a greater utilization value.

The present invention relates to a phosphorylated tau-gold nanoparticle complex, a method for manufacturing the same, and a drug screening method using same. The phosphorylated tau-gold nanoparticle complex manufactured according to the method for manufacturing a phosphorylated tau-gold nanoparticle complex of the present invention enables evaluation of how effectively a specific drug degrades phosphorylated tau aggregates and thus can be advantageously used for high-throughput screening of drugs for preventing or treating primary tauopathies.

The present disclosure provides a method of treating a neurodegenerative disease such as Alzheimer's disease in a subject. The method includes administering to the subject about 30 mg of AR1001 orally once daily, or a pharmaceutically acceptable salt thereof, and monitoring plasma phosphorylated tau 181 (pTau181) or phosphorylated tau 217 (pTau217) levels in the subject during treatment. The method may further include continuing AR1001 treatment if the subject's plasma pTau181 or pTau217 levels decrease during treatment. The subject may have mild cognitive impairment due to Alzheimer's disease or mild Alzheimer's disease dementia. The method may also include assessing the subject's cognitive function using one or more standardized tests during treatment. AR1001 may be administered in a pharmaceutical composition in the form of a tablet or capsule.

The invention discloses a phosphoric acid modified protein or polypeptide as well as a preparation method and application thereof in vaccines, and belongs to the field of protein phosphoric acid modification. The preparation method of the phosphoric acid modified protein or polypeptide comprises the following steps: carrying out nucleophilic reaction on a compound containing an aldehyde group and a phosphate group and protein or polypeptide to form Schiff base; and reducing the Schiff base by using a reducing agent, and carrying out post-treatment to obtain the phosphoric acid modified protein or polypeptide. After the phosphoric acid modified protein product prepared by the method and an aluminum hydroxide adjuvant are prepared into a vaccine and used for immunizing mice, the antibody titer is remarkably increased. The method is of great significance in improving immune effect, reducing inoculant times, saving antigen dosage and improving some people or improving the situation that immune response is weak under specific conditions.

The application discloses a method, system and device for detecting Tau protein based on machine learning of surface enhanced Raman spectroscopy, and the method comprises the following steps: S1, constructing a SERS signal probe with minimum initial Raman signal; S2, mixing a sample to be detected with the SERS signal probe, and collecting SERS spectra of the mixed system by using a Raman spectrometer; S3, pre-processing the collected SERS spectra, wherein the pre-processing comprises baseline correction, feature peak identification and minimum-maximum normalization processing in sequence, so as to obtain standardized spectral data; and S4, inputting the standardized spectral data into a machine learning regression model which is trained in advance, and automatically outputting a concentration value of Tau protein in the sample to be detected based on the input spectral characteristics by the machine learning regression model. The application can quantitatively detect phosphorylated Tau protein quickly, accurately and with low detection limit under the premise of keeping high specificity, and has high clinical conversion potential.

The invention discloses a synthesis method of a protein ligase mediated specific modified polypeptide, and belongs to the technical field of bioengineering. The problem that in the prior art, phosphorylated Tau protein is difficult to produce on a large scale is solved, Tau polypeptide is used for replacing full-length site specific phosphorylated Tau protein to serve as a positive standard substance in a clinical diagnosis kit, protein ligase is utilized for catalysis, segmented synthesis of polypeptide containing phosphorylated specific modification is achieved, and the specific phosphorylated Tau protein is obtained. Polypeptides from different sources can be efficiently and directionally connected under mild conditions, so that target proteins or polypeptide molecules with complete structures and various functions are obtained, and the peptide bond synthesis production efficiency is effectively improved.

The present application relates to a kit for detecting phosphorylated Tau protein p-tau 217 by luminescence immunoassay, and its use method and application. The kit includes R1 reagent, R2 reagent and R3 reagent. The R1 reagent contains luminescent microspheres and a first antibody coated on the luminescent microspheres, which has a first tag molecule and can specifically bind to p-tau 217. The R2 reagent contains a second antibody with a second tag molecule. The R3 reagent contains a first pairing molecule that can specifically bind to the first tag molecule. The first antibody and the second antibody can specifically bind to different epitopes of p-tau 217, and one first pairing molecule can bind at least two first tag molecules. The technical solution of the present application can improve the ability of the luminescent microspheres to receive reactive oxygen species generated by the photosensitive microspheres, enhance the detection sensitivity, and meet the needs of luminescence immunoassay for p-tau 217 in plasma.