60 results about "Phosphorylated proteins" patented technology

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 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 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.

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.

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 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.

Disclosed are methods for detecting and treating cancers (such as carcinoma, leukemia, lung cancer, colon cancer, central nervous system (CNS) cancer, melanoma, ovarian cancer, renal cancer, oral cancer, prostate cancer and / or breast cancer). The method may include receiving a sample from a patient (such as a sample from a biopsy, a sample from an extracellular vesicle, or a sample from a circulating tumor cell). The method may include determining whether a LY6C protein or a phosphorylated ANXA2 protein is detected in the sample. The method may include treating a patient for cancer by targeting the LY6C protein when the LY6C protein is detected (e.g., using photoimmunotherapy (PIT) or near infrared photoimmunotherapy (NIR-PIT)). or treating a patient for cancer by targeting an ANXA2 protein (e.g, using inhibitors, decoys, etc.) when the phosphorylated ANXA2 protein is detected.

This invention belongs to the field of antitumor drugs, disclosing a sulfated Polygonatum sibiricum polysaccharide, its preparation method, and its application in the preparation of drugs for treating liver cancer. The invention uses dried Polygonatum sibiricum rhizome as raw material, purifies it to obtain Polygonatum sibiricum polysaccharide, and then uses a sulfur trioxide-pyridine complex as a sulfation reagent to perform sulfation modification, thus preparing sulfated Polygonatum sibiricum polysaccharide. Sulfated Polygonatum sibiricum polysaccharide significantly improves water solubility, optimizes molecular spatial conformation and charge distribution, and induces cancer cell apoptosis by damaging mitochondrial membrane potential, increasing intracellular ROS levels, and downregulating the expression of key phosphorylated proteins in the PI3K / Akt / mTOR signaling pathway. In animal experiments, it effectively inhibited tumor growth without causing significant systemic toxicity in mice, demonstrating good in vivo biocompatibility. Therefore, it has important application value.

This invention provides a method for simultaneously introducing phosphorylated serine, phosphorylated tyrosine, and their non-natural amino acid analogs at specific sites in a protein, as well as a protein with dual phosphorylation modification prepared by this method and its applications.

The present application relates to the field of phosphorylated tau protein detection, and particularly relates to a method for improving the sensitivity of phosphorylated tau protein detection and a kit thereof.The present application provides a method for improving the sensitivity of phosphorylated tau protein detection, comprising: S101 obtaining a blood sample; S102 adding a detection reagent to the blood sample; S103 detecting the concentration of phosphorylated tau protein in the blood sample using a chemiluminescence method; wherein the detection reagent comprises an alkaline phosphatase polymer-antibody complex.The alkaline phosphatase polymer-antibody complex prepared by the present application has high yield, good stability, high coupling efficiency, significantly improved sensitivity, low background, good precision, strong anti-interference ability, smaller batch difference and lower non-specific adsorption, and exhibits significant advantages in detecting phosphorylated tau protein in blood samples.

Provided herein are compounds, methods, and compositions for determining whether a patient has a neurological disease or condition, comprising detecting the presence or cumulative mass thereof of a detectable target protein in a tissue or sample of the patient, such as the presence or cumulative mass thereof of amyloid beta protein or phosphorylated tau protein. The detection may comprise contacting the target protein with a compound described herein.

This invention relates to the pharmaceutical field, specifically to the application of tissue inhibitor metalloproteinases-2 (TIMP2) in the preparation of drugs for the prevention or treatment of traumatic brain injury. TIMP2 protein can increase the fall latency in rotarod experiments in mice with traumatic brain injury; improve the motor balance ability of mice with traumatic brain injury on a balance beam; improve neurological function impairment in mice with traumatic brain injury; and reduce Evans blue permeability in brain tissue. In brain microvascular endothelial cells (HBMECs), TIMP2 protein can participate in maintaining the integrity of the endothelial barrier, reversing the loss of expression and altered localization of the connectome complex induced in an in vitro traumatic brain injury model, and reducing luciferase leakage. This invention also provides the application of TIMP2 protein as an Integrin α3β1 ligand in the preparation of drugs for the treatment of central nervous system diseases caused by blood-brain barrier dysfunction. TIMP2 protein exerts its function by binding to the cell membrane receptor Integrin α3β1, regulating VE-Cadherin phosphorylation. TIMP2 protein shows promising application prospects in the treatment of blood-brain barrier dysfunction caused by traumatic brain injury.

Provided are a measurement method and a measurement kit which make it possible to suppress an albumin-dependent increase in background. A method for measuring phosphorylated tau protein according to the present disclosure includes a measurement step which includes: bringing a target sample into contact with a measurement reagent for phosphorylated tau protein in the presence of albumin and an amino group-containing compound, wherein the phosphorylated tau protein has a structure such that a threonine residue at position-217 is phosphorylated; and measuring the phosphorylated tau protein, which has a structure such that the threonine residue at position-217 is phosphorylated, in the sample.

The invention discloses a conjugate of phosphorylated Tau protein and magnetic beads as well as a preparation method and a kit of the conjugate, and relates to the technical field of immunoassay. The modified magnetic beads modified with azide groups and the modified anti-phosphorylated Tau protein antibody modified with DBCO groups are efficiently and directionally coupled in a click chemistry mode, and compared with a traditional physical adsorption or non-specific chemical coupling method, the method has the advantages of being high in coupling efficiency, high in binding stability, high in antibody activity retention rate and the like; compared with the prior art, the magnetic bead provided by the invention has the advantages that the capture efficiency and specificity of the magnetic bead on low-abundance p-Tau in blood can be obviously improved, the lower detection limit is as low as 0.3 pg / mL or below, excellent accuracy is shown in early AD diagnosis, subtle changes of p-Tau in the blood of an early AD patient can be accurately identified, and a reliable tool is provided for early screening, disease course monitoring and intervention effect evaluation of AD.

This invention discloses a single-sample multi-omics component extraction method for multi-omics joint analysis. The method includes: adding nucleic acid extraction solution to the sample, transferring it to a DNA adsorption column, centrifuging, and collecting the filtrate; washing the DNA adsorption column; eluting DNA; adding ethanol to the filtrate, transferring it to an RNA adsorption column, centrifuging, and collecting the filtrate; washing the RNA adsorption column; eluting RNA; and extracting proteins. This method integrates centrifugation column-based DNA / RNA co-extraction technology, a zinc ion-mediated protein precipitation and capture strategy, and an efficient Fe-IMAC phosphorylated peptide enrichment process. It achieves efficient protein precipitation and in-situ washing, thoroughly removing chemical components that interfere with mass spectrometry detection. It simultaneously achieves high-quality nucleic acid extraction and deep proteomics and phosphorylated proteomics coverage in a single sample, effectively solving the bottlenecks of traditional methods such as limited sample size, complex procedures, and poor reproducibility.

The invention discloses a new application of tripterine, namely application of the tripterine in preparation of bladder cancer resisting drugs, in-vitro cell experiments prove that the tripterine shows remarkable proliferation inhibition activity on various bladder cancer cell strains including cell strains UM-UC-3, TCCSUP and T24 and a cis-platinum drug-resistant strain T24CDDP, the median inhibitory concentration (IC50) of the tripterine is 0.2688-0.5870 mu M, and the tripterine shows remarkable proliferation inhibition activity on various bladder cancer cell strains including cell strains UM-UC-3, TCCSUP and T24 and a cis-platinum drug-resistant strain T24CDDP. The further mechanism research shows that the tripterine can regulate and control the cell cycle by down-regulating the expression levels of CDK1 and CDC5L proteins in bladder cancer cells and up-regulating the expression of phosphorylated p53 (p-p53) proteins at the same time, thereby playing a role in resisting bladder cancer; the invention provides a new effective candidate drug for clinical treatment of bladder cancer, especially cis-platinum resistant bladder cancer, and has important clinical application value.