125 results about "Phosphorylated proteins" patented technology

The invention relates to a light-activated chemiluminescence detection kit for phosphorylated Tau protein p-tau 217 as well as a use method and application of the light-activated chemiluminescence detection kit. The kit comprises a reagent R1, a reagent R2 and a reagent R3, wherein the reagent R1 comprises a luminous microsphere and a first antibody which is coated on the luminous microsphere, is provided with a first tag molecule and can be specifically combined with p-tau 217; the R2 reagent comprises a second antibody with a second tag molecule; the R3 reagent comprises a first pairing molecule capable of being specifically combined with the first tag molecule; the first antibody and the second antibody can be specifically combined with different epitopes of p-tau 217, and one first pairing molecule can be combined with at least two first tag molecules. According to the technical scheme, the capacity of the light-emitting microspheres for receiving active oxygen generated by the light-sensitive microspheres can be improved, the detection sensitivity is improved, and the light-activated chemiluminescence detection requirement of p-tau 217 in plasma is met.

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 present invention provides a phosphorylated Tau protein mutant and application thereof. The provided phosphorylated Tau protein mutant comprises at least one mutation substitution or deletion at E117, A145, G204, S305, A384, and L408 relative to the amino acid sequence SEQ ID NO.1 of the phosphorylated wild-type Tau protein, has improved thermal stability, and the thermal stability is significantly higher than that of the wild-type Tau protein during long-term storage. The phosphorylated Tau protein mutant can be used as a standard product and has greater utilization value.

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 present invention belongs to the field of biomedicine, and specifically relates to a combination and application of joint detection of biomarkers for Alzheimer's disease. The present invention provides an application of a biomarker group in detecting Alzheimer's disease, characterized in that the biomarker group includes: a) at least two of APOE4, GDF-15, UCH-L1 and PF4; or b) at least one of APOE4, GDF-15, UCH-L1 and PF4, as well as phosphorylated Tau protein and total Tau protein. The present invention also provides a kit for detecting Alzheimer's disease. The present invention contributes to a more effective detection and diagnosis of Alzheimer's disease, and provides a new path for the detection and prevention of AD.

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 relates to the technical field of biomedicine. The invention provides a phosphorylated protein marker for angiogenesis of glioma and application of the phosphorylated protein marker. Through a series of analysis such as iTRAQ quantitative phosphorylated proteomics, KEGG signal channel enrichment, a kinase spectrum, glioblastoma transcriptome high-throughput sequencing data and immunohistochemistry, it is determined that significantly different phosphorylated proteins and related signal channels can significantly affect neuroglioma angiogenesis; and the phosphorylated protein changes along with tumor progression, and is a phosphorylated protein marker and a potential therapeutic target for glioma angiogenesis.

The invention relates to the field of phosphorylated tau protein detection, in particular to a method for improving phosphorylated tau protein detection sensitivity and a kit thereof. The invention provides a method for improving phosphorylated tau protein detection sensitivity. The method comprises the following steps: S101, obtaining a blood sample; s102, adding the detection reagent into the blood sample; s103, detecting the concentration of the phosphorylated tau protein in the blood sample by using a chemiluminescence method; wherein the detection reagent comprises an alkaline phosphatase polymer-antibody compound. The alkaline phosphatase polymer-antibody compound prepared by the invention is high in yield, good in stability, high in coupling efficiency, low in background, good in precision, strong in anti-interference capability, smaller in batch difference and lower in non-specific adsorption, the sensitivity is remarkably improved, and the compound shows remarkable advantages in detection of phosphorylated tau protein in a blood sample.

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 belongs to the field of biological medicine, and particularly relates to application of phosphorylated Parkin protein in preparation of a medicine for treating neurodegenerative diseases carrying pathological alpha-synuclein aggregate characteristics. According to the application, research finds that the Parkin protein and the PINK1 protein have obviously different expression modes in a monkey brain in a physiological state, but the phosphorylation level of the Parkin depends on the expression of the PINK1 protein. Subsequently, it is further found that Parkin phosphorylation expression reduction and obvious aggregation of toxic pS129-alpha-syn occur in both the normal senescent monkey brain and the brain of a sporadic PD patient, and further, it is found through overexpression of wild Parkin that the S65 site phosphorylated Parkin can reduce accumulation of pS129-alpha-syn in the monkey brain, so that the accumulation of pS129-alpha-syn in the monkey brain can be reduced, and the accumulation of pS129-alpha-syn in the monkey brain can be reduced. And the mutant Parkin, which cannot be phosphorylated by PINK1, cannot reduce the aggregation of toxic pS129-alpha-syn. Therefore, the phosphorylated Parkin protein provided by the invention can be used for preparing medicines for treating neurodegenerative diseases carrying pathological alpha-synuclein aggregate characteristics.

To easily analyze a three-dimensional structure of a phosphorylated protein.SOLUTION: An analysis method according to an embodiment includes a three-dimensional structure prediction step and a classification step. In the three-dimensional structure prediction step, a three-dimensional structure of a phosphorylated protein is predicted by substituting phosphorylated serine and phosphorylated threonine in an amino acid sequence of the phosphorylated protein in which a target protein is phosphorylated with aspartic acid and glutamic acid, respectively, and inputting the amino acid sequence after substitution to a calculation model for three-dimensional structure prediction. In the classification step, the phosphorylated protein is classified on the basis of the three-dimensional structure obtained in the three-dimensional structure prediction step.SELECTED DRAWING: Figure 1

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 invention relates to the technical field of medicines, and discloses an experimental method of plumbagin in anti-non-small cell lung cancer drugs, the drugs comprise effective plumbagin and pharmaceutically acceptable carriers, and can induce expression of GPX4, so that the anti-non-small cell lung cancer drugs are obtained. The generation of ROS in non-small cell lung cancer subjects can be improved, the expression of STING and TBK1 phosphorylated proteins in the non-small cell lung cancer subjects can be increased, and meanwhile, the anti-tumor effect can be effectively improved when the compound is used together with a PD-1 inhibitor. A new drug treatment strategy is provided for clinical treatment of the non-small cell lung cancer, and theoretical and experimental foundations are laid for development of innovative drugs with proprietary intellectual property rights.

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.

To provide an immunogenic composition that induces an antibody to phosphorylated tau protein.SOLUTION: An immunogenic composition contains an antigenic peptide that induces antibody production to phosphorylated tau protein and carrier protein, where the antigenic peptide is a fragment of human tau protein including (1) YSpSPGpSPG (sequence number 2), (2) AKpSpTPpTAE (sequence number 5), (3) AKpSpTPTAE (sequence number 31), (4) AKpSTPpTAE (sequence number 32), (5) AKSpTPpTAE (sequence number 33) or (6) IVpYKpSPV (sequence number 24).SELECTED DRAWING: None

The invention provides an application of a GMFBS53 phosphorylated protein in a modified form of GMFBB as a biomarker in preparation of an MASH early diagnosis reagent. The invention further provides a kit capable of quantitatively or semi-quantitatively detecting the protein, and the kit contains an antibody capable of being specifically combined with the GMFBS53 phosphorylated protein. By detecting the expression level of the GMFBS53 phosphorylated protein, early diagnosis can be more efficiently and accurately carried out on the MASH condition of a subject, and an important basis is provided for early prevention and treatment of potential liver cancer patients. The kit provided by the invention can be suitable for early diagnosis of MASH.

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.

This application provides a monoclonal antibody against phosphorylated pTau217 protein at position 217. The heavy chain CDR amino acid sequence is H-CDR1:GYTFRNYW, H-CDR2:IYPGGGYT, H-CDR3:VVSDY; the light chain CDR amino acid sequence is L-CDR1:ESVEYSGISL, L-CDR2:AAS, L-CDR3:QQSRKVPST. This antibody can be used to prepare kits for diagnosing Alzheimer's disease, providing good sensitivity and specificity.

The present invention relates to a calculation-based method for analyzing phosphorylated proteins, comprising: a phosphorylated protein sequence prediction step (20) for predicting the probability of presence of each phosphorylation combination of a target protein using a calculation model, and selecting a phosphorylated protein in which the probability of presence is greater than or equal to a prescribed probability; a phosphorylated protein three-dimensional structure prediction step (30): respectively replacing phosphorylated serine and phosphorylated threonine in the phosphorylated protein sequence obtained by the phosphorylated protein sequence prediction step with aspartic acid and glutamic acid, and predicting the three-dimensional structure of the replaced protein by using a calculation model; and a classification and clustering step (40): classifying and clustering the phosphorylated proteins according to the three-dimensional structure obtained in the phosphorylated protein three-dimensional structure prediction step. According to the method, the three-dimensional structure of the phosphorylated protein can be predicted comprehensively, quickly and at low cost only by means of calculation of the AI model without an experiment, and then the phosphorylated protein capable of being used as a biomarker or a drug action target is screened based on the obtained three-dimensional structure.

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.