32 results about "Enzyme binding" patented technology

The invention relates to a UGT enzyme subtype substrate specificity prediction method and model. On the basis of a directional message passing neural network, graph structure characterization of a small molecule compound and features of specific protein binding sites of UGT enzyme are deeply fused, a bimodal prediction normal form of'molecule + protein binding sites' is designed, a deep learning model is constructed, conversion from compound center prediction to molecule-enzyme binding site comprehensive prediction is achieved, and the prediction accuracy is improved. And accurate classification prediction can be carried out on UGT enzyme substrates and non-substrates.

The invention relates to the technical field of biological medicine, in particular to an mRNA vaccine based on a protein degradation technology. The mRNA vaccine comprises an mRNA system and a lipid nanoparticle. The mRNA system refers to two mRNAs which can respectively encode an antigen-docking module and a recognition module-E3 ubiquitin ligase binding domain, the E3 ubiquitin ligase binding domain recruits an E3 ubiquitin ligase complex, the antigen is ubiquitinated and degraded into a peptide fragment by utilizing specific binding between the recognition module and the docking module, the peptide fragment is recognized by an MHC-I complex, and the E3 ubiquitin ligase binding domain is used for identifying the MHC-I complex. Then, the body immunity is activated, and the anti-tumor effect is realized. Besides, the invention provides a universal mRNA vaccine, that is, theoretically, different diseases can be treated only by replacing antigens in an mRNA system, the research and development time of the vaccine is greatly shortened, the research and development cost is reduced, a new thought is provided for tumor treatment, and the mRNA vaccine has a huge application prospect.

The invention discloses a PROTAC chimera for targeted degradation of an ROR [gamma] t receptor and application of the PROTAC chimera, the structure of the chimera is RW-L-Re, Re is a ligand capable of being combined with E3 ubiquitin ligase, L is a linking group covalently combined with at least one Re and at least one RW, and Rw is a target protein ROR [gamma] t binding ligand and is selected from one of the following structures. A series of PROTAC chimeras capable of degrading an ROR gamma t receptor in a targeted manner are designed and synthesized for the first time, a ternary complex is formed mainly by combining a target protein ligand and an E3 ubiquitin ligase ligand with POI and E3 ligase respectively, and then the POI is labeled with a ubiquitination tag and is further degraded by proteasome. Experimental results prove that the PROTAC chimera designed by the invention has excellent ROR [gamma] t receptor degradation activity, and can be used for preparing related drugs for treating autoimmune diseases or tumors.

The invention discloses a method for detecting Escherichia coli in food by combining Fe3O4 (at) Mo-CDs nano-enzyme with an aptamer, according to the method, near-infrared light response molybdenum-doped carbon dot modified Fe3O4 (at) Mo-CDs nano-enzyme is synthesized through a hydrothermal method, the Fe3O4 (at) Mo-CDs nano-enzyme has excellent NIR enhanced peroxidase activity and magnetism, 3, 3 ', 5, 5'-tetramethyl benzidine is oxidized to generate blue oxidized TMB, and the activity and magnetism of the NIR enhanced peroxidase are improved. After the surface of the Fe3O4 (at) Mo-CDs nano-enzyme is functionalized with an escherichia coli nucleic acid aptamer, the POD-like activity of the Fe3O4 (at) Mo-CDs nano-enzyme is inhibited, when Escherichia coli exists, the Fe3O4 (at) Mo-CDs nano-enzyme can be selectively combined with E.coli, the POD-like activity of the nano-enzyme is further inhibited, when the Fe3O4 (at) Mo-CDs nano-enzyme is used for detecting the E.coli in samples such as fruit juice and cakes, the detection limit is as low as 0.767 CFU / mL, and the Fe3O4 (at) Mo-CDs nano-enzyme has relatively high reliability and accuracy and can be widely applied to detection of E.coli. According to the invention, magnetic targeting separation and visual detection of E.coli are realized.

The application discloses a PROTAC compound with RORgamma t receptor targeted degradation and purposes thereof, which is a compound with a structure shown in a general formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutical composition thereof. e is a ligand capable of combining with an E3 ubiquitin ligase; the L is a linking group covalently combining at least one R e and at least one R W ; the R w is a target protein RORgamma t binding ligand; the application is based on the target point of RORgamma t and the PROTAC technology, and a series of RORgamma t-PROTACs are designed and synthesized for the first time. The mechanism is that the target protein ligand and the E3 ubiquitin ligase ligand are combined with POI and E3 ligase respectively, so as to form a ternary complex of "POI-PROTAC-E3 ligase". Then, the POI is marked with a ubiquitination label, and is degraded by a proteasome. The advantages of RORgamma t-PROTACs mainly include targeting of "undruggable proteins", small dosage, low toxicity, and difficulty in drug resistance. W -L-R e (I)

A compound of formula I or a stereoisomer, isotopic isomer, tautomer or pharmaceutically acceptable salt thereof: wherein a ligand that binds to Nef protein (NB) is covalently linked to a ligand that binds to E3 ligase cerebron (CB) via a linker (L).

This invention relates to the field of biopharmaceutical technology, and more particularly to a type of beeswax ( Fraxinus pennsylvanica This invention relates to a pollen allergen composition, its preparation method, and its application. It reveals that Chinese ash pollen contains more different allergenic proteins compared to European and American ash pollen. Therefore, using Chinese ash pollen as raw material, the resulting ash pollen allergen composition contains novel allergenic protein components that bind more strongly to the serum of Chinese ash pollen allergy patients, resulting in superior efficacy. This invention utilizes the ash pollen allergen composition to prepare a kit. This kit is highly sensitive, highly specific, and easy to operate. By changing the enzyme conjugate, it can be used for two purposes in one kit, achieving dual functions: detection of ash pollen allergen-specific IgE antibodies (in vitro allergen diagnosis) and quantitative detection of IgG antibodies (evaluation of desensitization therapy efficacy) in allergy patients.

This application relates to a source-aware activity ranking-based method for screening DPP-IV repressive peptides, comprising: constructing a training dataset containing positive samples and at least three levels of negative samples; extracting multimodal features of peptides, including at least sequence deep semantic embedding features and pocket structure features; explicitly aligning the above features using a cross-attention mechanism to generate fusion features; inputting the fusion features into the classification branch and ranking branch of a decoupled prediction architecture, outputting classification probability and activity ranking score respectively, wherein the ranking branch uses an independent ranking model and is trained based on preference pairs constructed from data within the same assay source; and performing virtual screening output by combining classification probability, activity ranking score, multidimensional confidence judgment, and ADMET safety screening results. This application achieves structurally interpretable characterization of substrate-enzyme binding modes and improves the statistical significance and generalization stability of peptide activity ranking under cross-laboratory batch effects.

The invention is applicable to the technical field of hydroxytyrosol preparation, and provides a method for preparing hydroxytyrosol by combining compound enzyme with molecular imprinting adsorption, which comprises the following steps: crushing fresh olive fruits, and collecting olive fruit pulp; putting the olive pulp into an enzymolysis tank, adding cellulose-pectin compound enzyme and citric acid, and performing enzymolysis; transferring the olive pulp subjected to enzymolysis into a microwave extraction tank, adding purified water, carrying out intermittent irradiation extraction, centrifuging after the extraction is completed, collecting supernate, and carrying out vacuum concentration; and adsorbing the concentrated supernate by using a chromatographic column filled with a hydroxytyrosol imprinted polymer, eluting with water and an ethanol solution in sequence, collecting ethanol eluent, and concentrating to obtain extract, namely the high-purity hydroxytyrosol. The problems that an existing fermentation method is low in yield and high in cost are solved. Generation of byproducts is reduced, and the separation and purification process is simplified; green and sustainable hydroxytyrosol industrial production is realized; and the purification period is shortened to be within 5 hours.

Provided herein are plasma kallikrein binding proteins such as antibodies binding to active plasma kallikrein and methods of using such proteins in treating hereditary angioedema.

Plasma kallikrein binding proteins and methods of using such proteins are described.

Provided herein are systems and methods for transposase-mediated insertion of an adapter construct into a target nucleic acid. A composition can include a nucleic acid molecule, comprising a plurality of barcode sequences and a plurality of capture sequences. The composition can further include a plurality of transpososomes coupled to the nucleic acid molecule, wherein each transpososome comprises an active transposase and a plurality of polynucleotides. The plurality of polynucleotides can include: (A) a pair of transposon end recognition sequences that are bound to the active transposase and (B) a polynucleotide comprising a binding sequence that is hybridized to a capture sequence of the plurality of capture sequences of the nucleic acid molecule.

Provided herein are novel Class 2 Type II and Type V CRISPR-Cas RNA-guided endonucleases such as Cas9 and Cas12 endonucleases and systems comprising the same. Methods of making and methods of using the same are also provided. Exemplary methods of use include modifying target DNA and detecting targeted DNA, which can be useful for therapeutic and diagnostic applications. Some diagnostic applications can take advantage of the collateral nuclease activity of the enzyme bound to the target sequence.

We describe compositions of DNA donor molecules compatible with various gene insertion methods, offering key advantages in delivery, nuclear localization, and immune evasion. The modified DNA donor molecules are predominantly single-stranded but include short regions of double-stranded DNA to reconstruct enzyme binding sites, and can be combined with other methods to enhance donor recruitment to the cell nucleus.

This invention relates to the field of monoclonal antibody technology, and in particular to OXA-48 enzyme monoclonal antibodies and their applications. The OXA-48 enzyme binding molecules provided by this invention exhibit high specificity, high biological activity, strong stability, small batch-to-batch variation, are unaffected by cell line degeneration, have high affinity for OXA-48 enzyme, and a titer exceeding 1:1,280,000, making them suitable for the detection or purification of OXA-48 enzyme products.

The application discloses a method for detecting escherichia coli in food by Fe3O4@Mo-CDs nano-enzyme combined with aptamer, wherein the near-infrared light responsive molybdenum doped carbon dot modified Fe3O4@Mo-CDs nano-enzyme is synthesized by a hydrothermal method, the nano-enzyme has excellent NIR enhanced peroxidase-like activity and magnetism, oxidizes 3,3',5,5'-tetramethylbenzidine to generate blue oxidized TMB, after the surface of the nano-enzyme is functionalized by escherichia coli nucleic acid aptamer, the peroxidase-like activity of the Fe3O4@Mo-CDs nano-enzyme is inhibited, when escherichia coli exists, the nano-enzyme can be selectively combined with the escherichia coli E. coli , and further inhibits the peroxidase-like activity of the nano-enzyme, when the nano-enzyme is used for detecting escherichia coli in juice, cake and other samples E. coli , the detection limit is as low as 0.767 CFU / mL, the nano-enzyme has higher reliability and accuracy, and the application realizes magnetic target separation and visual detection of escherichia coli E. coli .

The present application relates to the technical field of biology, and particularly relates to a method for stably improving horseradish peroxidase-labeled streptavidin titer. The method provided by the present application is to design streptavidin with different histidine tag lengths, and then couple with sodium periodate-oxidized horseradish peroxidase, so that high-titer horseradish peroxidase-labeled streptavidin can be obtained. The present application also discloses application of the enzyme-labeled conjugate in a magnetic microsphere platform streptavidin-biotin system. Experimental results prove that the method provided by the present application can stably prepare high-titer and low-background enzyme-labeled conjugate, and the enzyme-labeled conjugate can be widely applied in the streptavidin-biotin system.

The application belongs to the technical field of nano-enzyme sensing, and particularly relates to a three-mode phytic acid detection method based on a double-enzyme cascade reaction and application thereof. The detection method uses CAU-10-NH2 metal organic framework as a light-responsive nano-enzyme, combines the light catalytic oxidase activity of ALP and CAU-10-NH2, acquires signals through three modes of fluorescence spectrum, ultraviolet-visible absorption spectrum and photothermal imaging, and quantitatively analyzes the phytic acid content according to the signal change. Phytic acid inhibits the ALP activity, controls the generation amount of ascorbic acid, further influences the oxTMB content generated by CAU-10-NH2 catalyzing TMB oxidation, and realizes the synchronous change of fluorescence, colorimetric and photothermal signals. The application first discovers that CAU-10-NH2 has light catalytic oxidase activity, and only the -NH2 substituted derivative in the CAU-10 series (including CAU-10-H, -OH, -Br, -NO2, etc.) exhibits the activity, which expands the application of aluminum-based MOFs in the field of nano-enzyme sensing.

The application discloses a membrane strip incubation method for protein blotting and a detection method containing the same, and belongs to the technical field of immunoblotting. The membrane strip incubation method comprises the following steps: incubating a membrane strip by using a membrane strip incubation device, and placing the membrane strip into an incubation groove; adding buffer washing liquid and blocking liquid to the incubation groove, and blowing the liquid to flow to perform membrane blocking; adding a sample to be detected to the incubation groove, and blowing the liquid to flow to perform sample incubation; adding buffer washing liquid to the incubation groove, and blowing the liquid to flow to perform membrane washing; adding buffer washing liquid and enzyme combination to the incubation groove, and blowing the liquid to flow to perform enzyme combination; adding buffer washing liquid to the incubation groove, and blowing the liquid to flow to perform membrane washing; adding distilled water and color developing liquid to the incubation groove, shielding the membrane strip from light, and blowing the liquid to flow to perform light-proof color developing; adding distilled water to the incubation groove, and blowing the liquid to flow to perform color developing termination; and after each operation process is completed, liquid in the incubation groove is absorbed. The membrane strip incubation method ensures that the membrane strip is fully contacted with detection liquid and reacts.

The invention belongs to the technical field of vinasse functional development, and particularly relates to yellow wine vinasse polypeptide with Kokumi flavor enhancement and health promotion functions as well as a preparation method and application. The yellow wine vinasse polypeptide provided by the invention is prepared by the following steps: firstly, identifying three active peptides RFY, QMGR and AGRR which have strong binding capacity with a calcium sensitive receptor (CaSR) and glucolipid metabolism related enzymes from yellow wine vinasse by using high-throughput screening and molecular docking technologies, and synthesizing the three polypeptides through chemical synthesis methods such as a solid-phase synthesis method or a bioengineering method after obtaining amino acid sequences RFY, QMGR and AGRR. Sensory evaluation, an electronic tongue test and an in-vitro / in-vivo glycolipid metabolism related enzyme inhibition and oxidation resistance test are carried out, so that the three polypeptides are proved to be the multi-effect yellow wine vinasse polypeptides with Kokumi flavor enhancement and health promotion, and the technical problem of lack of the multi-effect yellow wine vinasse polypeptides in the prior art is solved.

The invention discloses a polybrominated diphenyl ether enzyme-linked immunosorbent assay rapid detection kit and a preparation method thereof.The kit comprises a BDE-47 standard substance, an enzyme conjugate, an antibody working solution, a substrate, a stop solution, a sample diluent I, a sample diluent II, a washing solution and a 96-hole elisa plate; by combining with a specific pretreatment method for water quality and aquatic products, high-sensitivity and rapid detection of BDE-47 is realized. The method is simple and convenient to operate, does not need a large instrument, has the detection time of about 90 minutes, has the detection limit of 1.5 ng / L (water quality) and the addition recovery rate of 80-120%, and is suitable for on-site rapid screening and grassroots popularization and application.

This invention provides an antibody or antigen-binding fragment thereof that binds to KPC enzyme and its application, relating to the technical field of monoclonal antibodies. The antibody or antigen-binding fragment that binds to KPC enzyme has a light chain CDR as shown in Seq_1-3, or as shown in Seq_11-13; and / or a heavy chain CDR as shown in Seq_4-6, or as shown in Seq_14-16. This antibody or antigen-binding fragment that binds to KPC enzyme exhibits high specificity and high affinity, alleviating the technical problem of poor efficacy of existing monoclonal antibodies that bind to KPC enzyme.

The present invention provides a linear multimeric biomolecule polymer wherein a biomolecule is bonded to a polyubiquitin scaffold formed of two or more covalently bonded ubiquitins, by obtaining, from a host cell, a biomolecule bonded with a ubiquitin C-terminal tag through recombinant expression, and polyubiquitinating the biomolecule in vitro in the presence of proteins involved in ubiquitination, E1 (activation enzyme), E2 (conjugation enzyme), and E3 (ligase), and a substrate. The polymer according to the present invention may be used in the separation and purification of a biomolecule, the separation of a target material that binds to the biomolecule, etc.

Plasma kallikrein binding proteins and methods of using such proteins are described.

This invention provides a nanozyme, its preparation method, and its application, belonging to the field of analytical chemistry detection technology. The nanozyme is vanadium oxide doped with a metal, in the form of nanoribbons. The metal is titanium, which is atomically loaded onto the vanadium oxide, with an atomic percentage of 1%-2%. The nanozyme of this invention possesses dual-enzyme activity similar to NADH oxidase and peroxidase. Utilizing its dual-enzyme activity in combination with natural enzymes, the amount of hydrogen peroxide is converted into a final measurement standard. Through the color change of a 3,3',5,5'-tetramethylbenzidine solution system, rapid qualitative and quantitative analysis of biomarkers for rare genetic metabolic diseases is achieved, providing a rapid, sensitive, and reliable colorimetric detection method.

The application belongs to the technical field of kits, and discloses an antinuclear antibody spectrum detection kit, a preparation method and a detection method. The detection kit comprises a detection film strip coated with a plurality of antigens in parallel, an enzyme combination liquid, a sample buffer and a substrate liquid. The antigens coated on the detection film strip include at least one of nRNP / Sm, Sm, SS-A, SS-B, Scl-70, PM-Scl, Jo-1, CENP-B, PCNA, dsDNA, nucleosome, histone, ribosome P protein, M2 and Ro52. The kit provided in the application can detect 15 kinds of antinuclear antibodies at the same time, has high accuracy, and is of great significance for assisting clinical diagnosis, clinical typing, disease observation, prognosis and treatment evaluation of autoimmune diseases.

The application provides an antibody or antigen-binding fragment thereof combined with IMP enzyme and application thereof, and relates to the technical field of monoclonal antibodies. The antibody or antigen-binding fragment thereof combined with IMP enzyme has light chain CDRs as shown in Seq_1-3 respectively, or as shown in Seq_11-13 respectively; and / or heavy chain CDRs as shown in Seq_4-6 respectively, or as shown in Seq_14-16 respectively. The antibody or antigen-binding fragment thereof combined with IMP enzyme has the characteristics of good specificity and high affinity, and alleviates the technical problem that the effect of the monoclonal antibody combined with IMP enzyme in the prior art is poor.

Disclosed are bacterial topoisomerase inhibitors with antibacterial activity, including against fluoroquinolone-resistant Staphylococcus aureus, comprising structural domains: a) a left-hand side (LHS) usually comprising a fused bicyclic or tricyclic ring system, b) a linker domain, and c) a right-hand side (RHS) comprising an aromatic or heteroaromatic ring, wherein the novel topoisomerase inhibitors do not contain a secondary amine at the start of the RHS, which is the enzyme-binding moiety.

The invention belongs to the technical field of in-vitro diagnosis, and particularly relates to a multi-joint detection anti-ganglioside antibody detection kit as well as a preparation method and application thereof. The kit comprises an enzyme-labeled microwell plate which is pre-coated with a glycolipid antigen and is subjected to sealing treatment, an anti-human IgG-HRP and anti-human IgM-HRP enzyme conjugate, a sample diluent, a washing solution, a TMB chromogenic substrate solution, a stop solution and a positive / negative reference substance. The solid-phase antigen comprises 12 kinds of single glycolipid antigens and 12 kinds of complex glycolipid antigens; the single antigen and the composite antigen are both subjected to solid phase presentation by adopting a membrane sample lipid mixed system formed by cholesterol-phospholipid-glycolipid, and the composite glycolipid antigen can be formed by co-assembly and co-coating of two glycolipids or by a membrane sample co-membrane co-assembly stabilization method. And each antigen is provided with IgG and IgM detection holes. The kit is standardized in process and quantifiable in result, and can be used for antibody repertoire auxiliary detection and analysis of related diseases such as immune-mediated peripheral neuropathy and the like.

The invention discloses an immobilized enzyme and a preparation method and application thereof.The immobilized enzyme comprises a carrier, protease and an immobilization layer, the immobilization layer comprises amino molecules and aldehyde molecules, the amino molecules comprise dendritic low polyetherimide, the amino molecules are formed through secondary hydroformylation of the aldehyde molecules, and the immobilized enzyme is rich in protease binding sites, high in enzyme activity and high in enzyme digestion efficiency; the method is suitable for a sample pretreatment process of chromatographic detection.