79 results about "Protein labeling" patented technology

The invention belongs to the technical field of gene engineering, and particularly relates to a biotin ligase mutant as well as a coding gene, an expression vector, a recombinant cell and application thereof. The amino acid sequence of the biotin ligase mutant is as shown in SEQ ID NO. 2. The biotin ligase (Bar A) mutant provided by the invention can be efficiently expressed in eukaryotes, the expression level of the biotin ligase (Bar A) mutant is obviously improved in mammalian cells compared with a wild type, and the biotin ligase (Bar A) mutant has good catalytic activity after expression, can effectively catalyze biotinylation of protein containing a biotin receptor sequence, and has good application prospects. The method has a wide application prospect in the fields of biotinylated protein preparation, and protein labeling, positioning, immunodetection, affinity purification, function research and the like based on a biotin-avidin system (BAS).

The invention discloses a probe Cvs-B-NHS based on a coumarin derivative as well as a preparation method and application of the probe Cvs-B-NHS. According to the fluorescent probe Cvs-B-NHS of the coumarin derivative, a coumarin mother nucleus and succinimide ester are organically combined, the excellent fluorescence characteristic of coumarin is reserved, efficient protein labeling capacity is achieved, high-sensitivity fluorescence signal output and specific labeling of protein are achieved, and the fluorescent probe Cvs-B-NHS has the advantages of being high in sensitivity and high in sensitivity. And the accuracy and reliability of protein detection and imaging are improved. The invention discloses a TDP-43 protein phase separation labeling method based on a Cvs-B-NHS probe, which provides a new means for researching the action mechanism of a TDP-43 (G335D) mutant in neurodegenerative diseases, and is beneficial to promoting the research progress in the field of neurodegenerative diseases. The application of the Cvs-B-NHS probe in evaluating the polarity change of the microenvironment in the protein liquid-liquid phase separation process in the cytoplasm environment caused by mitochondrial signal disturbance provides a powerful tool for the research of the protein phase separation microenvironment.

The invention belongs to the technical field of medicines, and discloses a near-infrared fluorescent probe taking an alpha, beta-unsaturated acryloyl ester unit as a response site of sulfydryl. The fluorescent probe has great advantages in the aspects of biological small molecule identification, protein marking and disease diagnosis, and is one of the most convenient and most popular tools for exploring a complex biological process. The fluorescent probe molecule capable of releasing red fluorescence after being activated by sulfydryl is obtained for the first time by connecting a fluorophore with an alpha, beta-unsaturated acryloyl ester unit. The fluorescent probe molecule disclosed by the invention can be used for specifically detecting sulfydryl in an organism, so that the aim of accurately monitoring the sulfydryl in a cell is fulfilled.

The disclosed systems and methods involve the semi-quantitative detection of disease-specific protein signatures in biological samples. The system may include a housing with a motor driven substrate holder on rails. The substrate holder is configured to be positioned at multiple locations along the guide rails. Imaging sensors capture images of portions of the substrate at these locations. The system detects markers on the substrate and determines the location of wells based on these markers. Based on the determined location of the wells, color values for each pixel in the wells can be measured. These values may be analyzed to detect disease-specific protein signatures in a variety of sample types.

The invention discloses an application of a biomarker in typing of polymorphic glioblastoma, and the biomarker is combined by selecting at least one of the following categories: at least one of a plurality of gene markers for identifying a proliferative type, a protein marker DLL3 and / or Ki67, a protein marker DL3 and / or Ki67, a protein marker DL3 and / or Ki67, a protein marker DL3 and / or Ki67, a protein marker DL3 and / or Ki67, and a protein marker DL3 and / or Ki67. At least one of a plurality of gene markers for identifying the neurosynaptic type, a protein marker GRMM3 and / or FGFR2; at least one of a plurality of gene markers for identifying the metabolic type, a protein marker P16; and at least one of a plurality of gene markers for identifying the immune regulation type, and at least one of protein markers CD44, C1R and Ki67. The application provides comprehensive features of EAS-GBM, and enhances understanding of occurrence, clinical stratification and treatment strategies of different ancestral tumors.

The present application relates to the technical field of fluorescent dyes, in particular to an asymmetric pentamethine cyanine compound. The compound comprises a structure as shown in formula I. The compound provided by the present application has good water solubility, near-infrared absorption, high fluorescence quantum yield and good molar absorption coefficient, and can be used in nucleic acid labeling, DNA sequencing, cell imaging, protein labeling or specific recognition of amino acids.

This invention provides a method and apparatus for splitting single-cell mixed sample sequencing data, relating to the field of biotechnology. The splitting method includes: capturing and sequencing single-cell suspensions using a single-cell platform; then performing reference genome alignment, cell identification, and gene expression level quantification on the sequencing data using Cellranger; splitting the cell data identified in step a into two groups of cell data for different sexes based on SNP locus information from the 1000 Genomes Project; and distinguishing the two groups of cell data from male or female samples based on the proportion of sex-specific genes expressed in the two groups of cell data. This splitting method eliminates the need for additional experimental operations such as protein labeling and genome sequencing, and can provide accurate and reliable data splitting even when individual SNP information is unavailable.

The utility model discloses a protein chromatography device. The protein chromatography device comprises a chromatography tank and a detection liquid injection pipe. The chromatography tank comprises a tank body, a first cavity and a second cavity, wherein the first cavity and the second cavity are arranged in the tank body. The bottom ends of the first cavity and the second cavity in the height direction of the tank body are flush. The first cavity and the second cavity are both used for filling protein column chromatography filler and a protein solution. And the detection liquid injection pipe penetrates through the wall thickness direction of the tank body and is communicated with the second cavity. And the detection liquid injection pipe is used for injecting detection liquid into the second cavity. The detection liquid is used for marking the target protein in the second cavity to display a separation and purification effect, and the separation effect of the target protein in the first cavity is reflected by the separation effect of the target protein in the second cavity. On the premise that the target protein in the first cavity is not affected, the separation condition of the target protein in the first cavity is reflected, and the convenience of knowing the separation and purification effect of the target protein in the chromatography process is improved.

The present invention belongs to the field of probe technology, and provides a trifunctional probe based on a photocrosslinking group, a preparation method thereof, and an application thereof. The method comprises the following steps: under a protective atmosphere, compound 1, compound 2, 1-hydroxybenzotriazole, dimethylformamide, carbodiimide, and N,N-diisopropylethylamine are mixed, and a condensation reaction is performed to obtain a trifunctional probe based on a photocrosslinking group. The probe provided by the present invention can be bonded with baicalein, TFAE, and scutellaria baicalensis medicated serum to form a corresponding probe, and compete with its original compound. Under excitation of different ultraviolet wavelengths, the probe can be cross-linked with small molecules and target proteins in traditional Chinese medicine, and accurately reflect the level of target protein labeling of the reactive group in the proteome through a "reporter group", and can also selectively enrich the labeled target protein, and achieve the identification of single compounds, compound combinations, and natural drug extract targets through subsequent experiments.

The invention relates to a protein crown labeling method based on a nano-enzyme ortho-position labeling technology, which comprises the following steps: in the presence of biotin phenol and H2O2, labeling a nano-protein crown based on the ortho-position labeling technology, and the nano-protein crown is formed by mixing nano-enzyme with peroxidase-like activity with protein. According to the invention, the inherent peroxidase-like activity of the nano-enzyme is utilized to develop a proximity labeling technology based on the enzyme activity, and protein crown protein and interacting protein thereof can be labeled in situ under the condition that the nano-particles form a protein crown; compared with a traditional protein crown centrifugal analysis method, the method can reflect the actual condition of the protein crown to a greater extent. According to the protein labeling method, labeling of the soft crown protein can be achieved, protein enrichment can be carried out through the label, and a thought method for collecting and analyzing the soft crown protein is provided.

The invention relates to a nitroimidazole group meso-substituted heptamethine cyanine dye as well as a preparation method and application thereof. According to the dye, a nitroimidazole group is introduced into the middle position of a heptamethine cyanine parent nucleus, so that the dye has excellent absorption characteristics in a near-infrared region, the excitation wavelength is 730-780 nm, the emission wavelength is 800-860 nm, and the dye has good tissue penetrating power. The nitroimidazole group can effectively quench fluorescence in an excited state through a light-induced electron transfer effect, energy is promoted to be released in a non-radiative transition mode, the photothermal conversion efficiency is remarkably improved, and meanwhile high light stability and low fluorescence quantum yield are kept. The dye can be further prepared into a nano preparation and shows excellent biocompatibility and tumor targeting ability. The compound has wide application prospects in the fields of non-diagnostic or therapeutic protein labeling, bioimaging, photothermal therapy and the like, is especially suitable for preparing efficient photosensitive reagent drugs for tumor photothermal therapy, and provides a new strategy for design of near-infrared photothermal diagnosis and treatment materials.

Provided are diagnostic and treatment methods,based on plasma protein markers for neurodegenerative diseases such as mild cognitive impairment (MCI) and Alzheimer's Disease (AD),as well as kits for diagnosing and / or treating these condition.Machine learning systems and methods for assessing the risk for a subject having a neurodegenerative disease based on measured protein marker levels are also provided.

The invention discloses an ultra-stable red fluorescent protein. The fluorescent protein disclosed by the invention is a protein with an amino acid sequence SEQ ID No.4. Experiments prove that the fluorescent protein has excellent electron microscope sample preparation resistance, can retain more fluorescent signals after osmic acid treatment, and can tolerate embedding of Epon resin; meanwhile, the fluorescent protein has excellent thermal stability, and can retain more fluorescent signals at 90 DEG C; in addition, the fluorescent protein has high chemical stability and light stability. Based on the related properties of the fluorescent protein, the fluorescent protein can be used in the fields of protein labeling, various types of fluorescence imaging and super-resolution photoelectric correlated imaging, and is used for tracking the structural and morphological changes of samples such as proteins, subcellular organelles, local cell regions and cells; the kit can be used for packaging related viruses or transgenic animals, marking specific proteins, specific organelles or specific cells, and realizing rapid transparent tissue imaging and expansion super-resolution imaging.

The invention belongs to the technical field of biological medicines, and particularly relates to sulfonium salt polypeptide or protein as well as a preparation method and application thereof. Polypeptide or protein containing cysteine and a sulfonium salt compound are used as raw materials, and the sulfonium salt compound is used for alkylation of cysteine at a specific site of the polypeptide or protein to prepare the sulfonium salt polypeptide or protein. The sulfonium salt polypeptide or protein is used as a probe for covalently labeling tryptophan in a lysine methylation reader protein recognition pocket. According to the method disclosed by the invention, different types of sulfonium salt polypeptides and protein probes can be simply, conveniently and efficiently prepared, and the designed annular sulfonium salt with a novel skeleton has an obvious improvement on the polypeptide or protein labeling yield of reader protein. The prepared sulfonium salt probe not only can be used for identifying lysine methylation modified reader protein and finding a therapeutic target for disease-related pathways, but also can be used for modifying tryptophan at a specific site of an antibody-coupled drug and provides a powerful tool for targeted therapy research of tumors.

The present invention relates to a method, system and device for predicting immune microenvironment scores based on tissue mass spectrometry imaging. The method comprises: obtaining an image of mass spectrometry imaging of tumor tissue of a patient to be tested; obtaining single cell morphology information and protein marker information in the image according to the image of mass spectrometry imaging of the tumor tissue, classifying the cell population according to the single cell morphology information and protein marker information in the image, and obtaining cell subpopulation data; inputting the cell subpopulation data into an immune microenvironment scoring model to obtain the immune microenvironment score of the patient to be tested. The present invention develops an auxiliary diagnosis and treatment system that can predict immune microenvironment scores to accurately identify patients who benefit from immunotherapy, which is very important for practical clinical applications.

The invention discloses a space-time marking method for a living cell endoplasmic reticulum proteome, and belongs to the field of proteomics. The technical problem to be solved by the invention is how to mark endoplasmic reticulum proteins. The space-time labeling method for the living cell endoplasmic reticulum proteome comprises the following steps: treating a cell to be detected by using a metal iridium complex Ir3 and a thiomethylene quinone precursor probe to obtain the cell treated by the Ir3 and the probe; the thiomethylene quinone precursor probe is a compound or a derivative thereof obtained by connecting biotin and a thiomethylene quinone precursor with a linking group; and irradiating the Ir3 and the cells treated by the probe with blue light to realize space-time labeling of the endoplasmic reticulum proteome of the living cells. The method disclosed by the invention can be used for performing space-time labeling on the endoplasmic reticulum proteome of the living cell, has the characteristics of high specificity and in-situ protein labeling, and has nanoscale space-time resolution.

The present invention discloses an anti-pigeon adenovirus Hexon protein monoclonal antibody, the anti-pigeon adenovirus Hexon protein monoclonal antibody is monoclonal antibody 1H5-8-14-5, wherein the amino acid sequences of the heavy chain variable region and the light chain variable region of monoclonal antibody 1H5-8-14-5 are shown in SEQ ID NO.3 and SEQ ID NO.4, respectively. The present invention also discloses a double antibody sandwich ELISA diagnostic kit for detecting pigeon adenovirus antigens, the kit comprising an enzyme labeling plate coated with an anti-Hexon protein polyclonal antibody, Hexon protein, HRP-labeled anti-pigeon adenovirus Hexon protein monoclonal antibody, a negative control, a color developing solution, a washing solution, and a stop solution. The kit of the present invention can effectively detect pigeon adenovirus, provides a new technical means for pigeon adenovirus prevention and control, and the detection result is accurate and specific, with good application value.

A protein marker panel for screening clinical futile recanalization (FR) in patients with large vessel occlusion-induced acute ischemic stroke (LVO-AIS) and use thereof are provided. The protein marker panel includes GUCA2A, CCL14, STAB1, and VSIG4. A combined diagnostic value of the protein marker panel is high, and receiver operating characteristic (ROC) curve analysis demonstrates an efficacy in distinguishing and screening FR patients, with an area under the curve (AUC) of 0.959, indicating desirable sensitivity and specificity. Such a protein marker panel shows promise as a significant indicator for clinical decision-making in FR evaluation.

The present invention relates to a protein marker fluorescence detection method based on cyclodextrin-adamantanized iridium complex host-guest assembly labeling, which belongs to the application technology of organic iridium complex hydrogenation catalysis technology in the field of biological fluorescence sensing. In order to solve the numerous biocompatibility problems such as easy thiol deactivation faced by organic iridium complex hydrogenation catalysts in biological applications, the present invention adopts the following technical scheme: (1) preparation of a nano material M co-labeled with an organic iridium complex with high catalytic hydrogenation activity and a labeled antibody, and successfully solves the irreversible problem of easy thiol deactivation of the metal iridium complex in protein labeling detection by regulating the modification sequence of the labeled antibody, β-cyclodextrin, a blocking agent and adamantane functionalized metal iridium complex during the material assembly process; (2) coating an antibody A on a carrier; (3) adding a detected antigen biological protein marker; (4) adding the organic iridium complex and the nano material M co-labeled with the labeled antibody; and (5) adding a fluorescent detection liquid containing a hydrogenation regulation probe, wherein the iridium complex can catalyze the probe molecule to produce a fluorescence signal change, and quantitative detection of the biological protein marker to be detected can be achieved by testing the fluorescence intensity change of the detection liquid. This technology realizes the rapid and sensitive analysis of biological protein markers and has the advantages of simple operation and good thermal stability.

The invention discloses a rapid protein labeling kit which comprises a freeze-drying agent, an initiator and a terminator, and the freeze-drying agent is obtained by dissolving a micromolecular to-be-labeled substance in a buffer solution and freeze-drying; the starter comprises the following components: an alkaline buffer solution with the pH value of 8.5-9.5 and the concentration of 0.2-0.5 M; 10%-20% of an organic solvent; 0.05% to 0.2% of a preservative; the terminator comprises the following components: 0.1 to 0.5 M of amino acid with free amino groups; 1%-5% of bovine serum albumin; and 0.05%-0.2% of a preservative. The kit is used for protein marking, the operation is simple, the marking efficiency is high, the stability of a marker is good, and the kit can be stored for a long time.

The application discloses a fluorescent probe for adjacent thiol protein labeling and bio-imaging and application thereof, which is used for in-vitro labeling of adjacent thiol proteins and bio-imaging of endogenous adjacent thiol in living cells. Based on the active dimethyl sulfide site on cyclopentanone, a series of adjacent thiol protein recognition probes (IDAs) with high specificity, high stability and biocompatibility are constructed and screened, so that in-vitro fluorescent detection of adjacent thiol proteins and in-situ fluorescent labeling of adjacent thiol proteins in cells are realized.

The present invention relates to an integrated centrifugal device and method for analyte processing, particularly for the separation, enrichment, labeling, and analysis of nanovesicles such as extracellular vesicles (EVs). The device leverages a compact, rotatable substrate to execute multi-step assay protocols that may include chromatographic separation, centripetal fluid transport, immuno-capture, and protein labeling, all under low-speed centrifugal conditions. The device architecture enables automated, hands-free operation without requiring ultracentrifugation or specialized personnel. Fluidic modules including sedimentation, chromatography, displacement, and multiple assay chambers may be functionally integrated through a series of valves, channels, and immiscible liquid-based displacement mechanisms, facilitating precise sample handling, minimal loss, and multiplexed analysis.

The invention provides a method for carrying out histological staining by using a protein marker and application of the method in distinguishing sieve molecules and parasporal cells, and belongs to the technical field of biological histology. The invention discloses a method for distinguishing early tubular cells of plant phloem sieve molecules and paragrams, which can distinguish two symbiotic tubular molecules, namely sieve molecules and paragrams, of the phloem by utilizing an apparent protein marker and adopting a histological staining method before cell nucleus of a specific structure such as the sieve molecules disappears by utilizing whether the cells are colored or not; meanwhile, the specific position of interaction between the methylated histone and the paragraph can be distinguished according to the coloring position and the coloring depth, the development degree of the paragraph can be evaluated, and the sensitivity and the precision are high.

The application relates to the field of chemiluminescence immunoassay, in particular to a water-soluble acridine sulfonamide carboxylic acid label, a preparation method, application and kit, wherein in some embodiments, the water-soluble acridine sulfonamide carboxylic acid label is coupled by one-step reaction of acridine sulfonamide and a water-soluble glycine tripeptide, and the synthesis is simple and the yield is high. The water-soluble acridine sulfonamide carboxylic acid label not only increases water solubility, but also prolongs a connecting arm, and solves the water solubility problem of a protein label.

The application discloses a method for preparing aromatic benzyl propargyl ketone by light induction and biological application, belongs to the technical field of organic synthesis and biological medicine, and converts aromatic benzyl propargyl alcohol into aromatic benzyl propargyl ketone under the condition of light induction; the change of the functional group before and after light induction is that the hydroxyl alkyne without reaction activity is converted into the carbonyl alkyne with click reaction activity, fluorescence enhancement can be realized, and the method can be used for imaging and tracing; and the aromatic benzyl propargyl ketone can react with a compound containing an amino group or a biological macromolecule to realize the killing of tumor cells. The method uses light as a starting switch, the reaction condition is simple, controllability is good, no additional catalyst is needed, and the method is easy to mass-produce, so the method has wide application prospects in polypeptide or protein labeling, anti-tumor and the like.

The invention provides a protein marker group for detecting myeloid cells, application, a kit and a detection method. The protein marker group comprises a mononuclear-phagocyte line antibody marker, a neutrophil antibody marker, an antigen-presenting cell antibody marker, analogues of the antigen-presenting cell antibody marker and the like. The method can accurately describe the states of tumors and immune cells in a tumor microenvironment, provides important reference information for accurate diagnosis of the tumors, and also has important significance for immunotherapy of the tumors, including the proportion and distribution of immune cell subpopulations, especially the spatial relationship between immune cells and tumor cells, and the like. All the results are important contents for tumor prognosis and prediction research.

The present invention relates to the field of protein labeling or tracing, and particularly to a composition and method for labeling or tracing a target protein. The method first analyzes the structure of the target protein and identifies a loop sequence on the protein surface that faces outward. Then, through molecular cloning methods, a small tag (such as an HA tag) with flexible amino acid linker peptides at both ends is inserted into the middle of the loop sequence. Finally, a nanobody fused to the small tag and fused to a fluorescent protein is expressed. This method utilizes the nanobody's ability to specifically recognize the target protein surface tag to achieve intracellular tracing of the target protein.

The present invention discloses a Mcl-1 protein labeling probe based on click chemistry, a preparation method and an application thereof, and belongs to the field of synthesis and detection technology. The present invention utilizes the highly selective, efficient and rapid reaction characteristics of click chemistry to design a Mcl-1 protein labeling probe comprising a click reaction module with a Mcl-1 protein-specific targeting group and a click reaction module with a fluorescent marker. The two modules can be rapidly combined through a click chemistry reaction; the structural formulas of BCN-ID and Tz-BODIPY are as follows: #imgabs0# The structural formulas of BCN-ID and Tz-BODIPY after combination through a click chemistry reaction are as follows: #imgabs1# The two click reaction modules can be rapidly combined through a click reaction. This "turn-on" type fluorescence enhancement probe based on click chemistry has a novel structure and good selectivity and can be applied to imaging studies of intracellular Mcl-1 proteins.

The invention belongs to the technical field of medical analysis and detection articles, and particularly relates to a microporous polypeptide chip, a preparation method and application thereof and a label-free affinity screening method of targeted polypeptide. According to the microporous carrier chip provided by the invention, the same or different polypeptides can be subjected to a bonding reaction with chemical groups on the inner surfaces of the micropores in the hollow micropores, and the obtained microporous polypeptide chip is a trace, efficient and controllable polypeptide chip; the polypeptide of the microporous polypeptide chip provided by the invention does not need to be subjected to signal labeling by using a fluorophore or other labeling technologies, and when the microporous polypeptide chip provided by the invention is used for carrying out a targeted affinity screening test, protein labeled by fluorescence or other labels is not needed, so that a polypeptide compound labeling or protein labeling process is omitted, and the cost is reduced. Meanwhile, the dosage of the target protein is small, the screening operation process is greatly simplified, the screening efficiency is improved, the screening cost is saved, and negative effects generated by signal labeling groups such as fluorescence are eliminated.

Provided are a fluorescent probe, a preparation method therefor and a use thereof. The fluorescent probe sensitively and specifically responds to viscosity, and can be used for the specific fluorescence labeling of proteins as well as in the quantification, detection or kinetic study of proteins and the imaging of cells, tissues and living bodies.