29 results about "Chemical labeling" patented technology

The determination of chemical mixture components is vital to a multitude of scientific fields. Oftentimes various spectroscopic methods are employed to decipher the molecular composition of these complex mixtures. The sheer density of spectral features of different molecules present in such observations may make unambiguous assignment to individual species using these methods challenging. Yet, components of a mixture are commonly chemically related due to environmental processes or shared precursor molecules. Therefore, along with investigating the spectroscopic signals, analysis of the structural and chemical relevance of a molecule is an important consideration when determining which species are present in a mixture. Machine-learning molecular embedding methods are used with a relevance module to determine the likelihood of a molecule being present in a mixture based on the other known species, chemical priors, and spectroscopic information. By incorporating this metric, the mixture components can be identified with extremely high accuracy (∼ 97%).

The invention discloses an engineered humanized fluorescent direct-labeled antibody for identifying an SND1 phase separation signal and preparation, the fluorescent direct-labeled antibody is SND1-Cy5, and the SND1-Cy5 is obtained from an SND1 antibody and Cy5 by using a light-controlled biological orthogonal chemical labeling technology. According to the invention, through humanized transformation, the binding specificity of the antibody is optimized. The preparation process of the antibody ensures the consistency and stability between batches, and solves the problems of large difference between batches and complex liquid storage components of the existing antibody. The SND1-Cy5 direct-labeled antibody disclosed by the invention can be used for effectively identifying SND1 phase separation signals formed in cells under different stress conditions. According to the method, the defects in the prior art are overcome, the specificity, the sensitivity and the stability of the SND1 antibody are remarkably improved, the challenge of phase separation signal detection is successfully dealt with, and a powerful tool is provided for researching cell stress response and a tumor drug resistance mechanism.

The invention relates to an anti-mouse IgG nano antibody and alkaline phosphatase fusion protein as well as a preparation method and application thereof. The anti-mouse IgG nano antibody and alkaline phosphatase fusion protein sequentially comprises an anti-mouse IgG nano antibody, a connecting peptide and shewanella alkaline phosphatase from an amino terminal to a carboxyl terminal. The fusion expression of the anti-mouse IgG nano antibody and shewanella alkaline phosphatase in escherichia coli is designed, so that various defects of a chemical labeling method can be avoided, a high-purity and high-specific activity enzyme-labeled antibody can be prepared, the preparation period of a second antibody can be shortened, the preparation process of the second antibody can be simplified, the stability of batches can be improved, and the dependence on experimental animals can be reduced. The prepared anti-mouse IgG nano antibody and alkaline phosphatase fusion protein can be applied to the field of immunodetection, such as a chemiluminescence immunoassay method, an enzyme-linked immunosorbent assay method, an enzymatic fluorescence immunoassay method, an immunoblotting method and the like.

The present invention belongs to the fields of molecular biology and clinical medicine, and relates to a nucleic acid aptamer targeting IGF-1R and its application. The nucleic acid aptamer targeting IGF-1R includes any one of the nucleotides shown in SEQ ID NO.1 to SEQ ID NO.3; or any derivative of a nucleic acid aptamer with the same function obtained by chemical modification, chemical labeling or base change on the basis of SEQ ID NO.1 to SEQ ID NO.3. The nucleic acid aptamer of the present invention can be synthesized in large quantities in vitro, is cheap, has good stability, high affinity, is convenient for storage, and is easy to edit; at the same time, due to its small molecular weight, it is not easy to cause immune reactions, and the chemical stability of single-stranded DNA is good, and denaturation and renaturation are reversible, making it easy to store for a long time and transport at room temperature.

The invention relates to ribozyme for labeling biotin on target RNA (Ribonucleic Acid) as well as a screening method and application thereof. The RNA labeling technology is one of core tools for modern molecular biology research and clinical diagnosis. A specific marker is covalently linked to a target RNA molecule, so that accurate tracking, efficient detection and functional analysis of RNA can be realized. A biotin-streptavidin system becomes one of the technical routes which are most widely applied due to extremely high affinity and signal amplification capability of the biotin-streptavidin system. At present, common biotin-labeled RNA technical methods mainly comprise a chemical solid-phase synthesis method, an enzymatic method and a chemical labeling method. However, the methods have the limitations of poor site specificity, low efficiency, complex manipulation and the like in the use process. The engineering ribozyme for catalyzing RNA biotinylation is obtained by taking 12: 0 biotin coenzyme A as a biotin donor through an in-vitro screening technology. The ribozyme provided by the invention can efficiently and specifically carry out biotin labeling on target RNA, the reaction process is simple, and the labeling activity is up to 90% or above.

This invention belongs to the field of compound technology and provides a DNA-intercalating fluorescent quenching compound, its preparation method, and its application. This invention provides a DNA-intercalating fluorescent quenching compound that exhibits absorption in the visible light region and certain DNA sequence selectivity. The DNA-intercalating fluorescent quenching compound provided by this invention can be further used to design fluorescence analysis systems for restriction endonuclease EcoRV and DNases. The DNA-intercalating quenching compound can achieve relevant fluorescence analysis by combining with fluorescent molecules in a covalent-free manner, reducing the need for chemical labeling steps, thereby improving efficiency and reducing detection costs, and showing good application prospects in related analyses.

The invention discloses a resolution positioning analysis method for an 8-oxo-2 '-deoxyguanine modified single base in DNA (Deoxyribonucleic Acid), and belongs to the technical field of biology. The method comprises the following steps: firstly, extracting genome DNA, ultrasonically breaking, then denaturing into single-stranded DNA, and enriching the single-stranded DNA containing 8-oxo-dG by using an antibody; adding a 3 ' / 5' universal sequencing joint to the enriched single-stranded DNA; fixing the amino-modified extension primer to the carboxyl-modified magnetic bead to obtain an extension primer fixed by the carboxyl-modified magnetic bead; extension is carried out through an extension primer fixed by a carboxyl modified magnetic bead, and 8-oxo-2'deoxyguanine-thymine in DNA polymerase mediation enrichment single-stranded DNA is converted, so that a primer extension product fixed on the magnetic bead is obtained; purifying a primer extension product fixed on the magnetic beads; and carrying out PCR amplification on the primer extension product, and carrying out enzyme digestion or sequencing analysis. According to the method, the reaction system is mild, the defect that DNA oxidation modification is possibly artificially introduced in an existing chemical labeling method is avoided, and Ggt is passed; t conversion is carried out, so that the 8-oxo-dG can be modified, positioned and analyzed more directly and accurately.

The present application belongs to the technical field of analysis and identification of fermented food, and particularly relates to a method for distinguishing and identifying fermented food of different processes based on chemical isotope labeling liquid chromatography-mass spectrometry (CIL-LC-MS) technology analysis of carboxylic acid. In order to accurately and simply identify the type and content of carboxylic acid, and thus based on carboxylic acid panorama analysis, efficiently and conveniently distinguish and identify fermented food of different processes, the present application introduces CIL-LC-MS technology to analyze carboxylic acid and thus distinguish and identify fermented food of different processes. The technology can significantly enhance the response value and detection sensitivity of carboxylic acid through chemical labeling strategy, and effectively overcome the ion suppression interference of the matrix by using isotope internal standard, so as to realize accurate quantification. Combined with its inherent three-level identification process, the present application can efficiently and conveniently distinguish and identify fermented food of different processes.

The invention relates to the technical field of biological detection, in particular to a dry-state hydrogel-based plasma-Fabry-Perot cavity biological detector as well as a preparation method and application thereof. Compared with ELISA and other immunological detection methods depending on chemical tags and signal amplification, the method does not need a signal amplification process and shortens the detection time. The integrated plasma interlayer thickness sensing structure provided by the invention realizes direct signal reading after sample incubation is finished. According to the invention, the hydrogel is used as a sensing interlayer, and the hydrogel is used as a three-dimensional medium, so that a larger thickness change can be generated in a spatial scale. The biological detector can still keep relatively high specificity in a complex protein liquid phase environment (serum). The device has the structural characteristics of chip formation and miniaturization, and can be used for detecting various types of high-throughput proteins at the same time.

The invention provides a cross-linking chemical labeling method for exosome membrane proteomics analysis, and belongs to the field of exosome membrane proteines.The method comprises the following steps that a cross-linking labeling efficiency evaluation system is established, the cross-linking labeling density value and the spatial structure selectivity value of a cross-linking labeling reagent within the time range of 0-3600 seconds are measured, and the cross-linking labeling efficiency evaluation system is established; obtaining a standard crosslinking label density value and a standard spatial structure selectivity value in a standard crosslinking state; calculating a first judgment parameter and a second judgment parameter of the cross-linking labeling reagent to be detected; when the first judgment parameter is greater than or equal to 0.8 and the second judgment parameter is greater than or equal to 0.7, adding the to-be-detected cross-linking labeling reagent with the concentration of 1-5 mmol / L into exosome suspension with the concentration of 1 * 10 < 12 > pieces / L; and determining the distribution uniformity value of the cross-linking mark and the degree of completion of the cross-linking reaction in the exosome suspension. The method can solve the technical problem of unstable marking efficiency in the prior art.

The utility model belongs to the technical field of optical biosensors, and particularly relates to an enhanced Fabry-Perot microcavity sensor for ultralow-concentration biological detection. The sensing microcavity comprises a square capillary quartz tube, wherein high-reflectivity films are plated on the upper surface and the lower surface of the square capillary quartz tube; a liquid core area of the square tube is a micro-flow channel, and a polyethylene glycol polymerization promoting biomolecule solution is introduced into the micro-flow channel, so that the Fabry-Perot microcavity biosensor is obtained. According to the utility model, the square capillary quartz tube provides high-precision parallelism between the two reflectors, so that the diffraction and walk-off loss can be reduced, the mode volume in the cavity can be reduced, the light energy density can be increased, the strong interaction between light and substances can be realized, and the quality factor can be improved; polyethylene glycol can promote the polymerization of biomolecular protein, enhance the reaction of an antigen-antibody compound, and further promote the aggregation of an immune compound. The biosensor is simple in structure, convenient to prepare, good in repeatability and stability, high in response speed and free of surface modification and chemical labeling.

Methods and kits for detecting analytes (such as a plurality of analytes) in a sample utilizing signal amplification (such as DMP-based chemical tag signal amplification) are provided.

The invention belongs to the technical field of biology, and particularly relates to a method for detecting and enriching glycosylated RNA (Ribonucleic Acid) based on in-vitro enzyme chemical labeling of alpha-2, 3, alpha-2, 6 type sialyltransferase. According to the application, sialidase is firstly used for digesting sialic acid at the tail end of a naturally generated glycoRNA sugar chain, and sialidase digestion is not needed for an RNA sample which does not have sialic acid at the tail end of the glycoRNA sugar chain but has potential sialic acid modification sites; sialic acid with a labeled group is transferred to a potential sialic acid modification site at the tail end of a glycoRNA carbohydrate chain by using alpha-2, 3, alpha-2, 6 type sialyltransferase, such as PmST1, PmST3, Ph2, 3siat and Pd26ST, so that glycoRNA with a detectable group can be directly obtained, or the detectable group is further introduced through subsequent reaction, so that subsequent signal detection or enrichment can be carried out; the RNA part can be further sequenced, and the glycosyl part can be further subjected to mass spectrum identification. The application provides a very efficient, simple and convenient detection means for detection and enrichment of sialylated glycoRNA, has the characteristics of high specificity and high sensitivity, and has a wide application prospect.

The invention discloses an unmarked quantum dot electrochemical luminescence sensing electrode for detecting mycotoxin as well as a preparation method and application thereof. The preparation method comprises the following steps: preparing an interdigital electrode pair with two groups of independent detection channels on a glass substrate; self-assembling cysteamine on the surface of the gold working electrode to form an amination interface; sequentially carrying out covalent immobilization on the ZnCdS (at) ZnS quantum dots and different fungaltoxin aptamers through amidation reaction; and finally, sealing by using bovine serum albumin. The prepared sensing electrode has a unique complementary segmented interdigital structure, comb tooth parts of the gold working electrode and the platinum counter electrode are arranged in a staggered and non-contact manner, and annular parts are arranged in a concentric circle manner, so that electric field interference between channels is effectively avoided. The invention further provides application of the electrode, chemical labeling is not needed in the whole process of the method, high-sensitivity, high-specificity and parallel in-situ detection of high-harm mycotoxins such as ochratoxin A and zearalenone is achieved, the integration level is high, and the electrode is suitable for on-site rapid screening of food safety.

This invention relates to the field of biodetection technology, specifically disclosing a multiplex fluorescent staining kit for tissue samples. The kit includes an antibody dissociation buffer, a signal amplification deposition solution, a fluorescent labeling reagent, a tissue-protective imaging medium, and an autofluorescence background inhibitor. This invention utilizes a non-destructive elution technique achieved through the synergistic effect of a low-pH buffer and a directional electric field. It leverages the positively charged nature of antibodies under acidic conditions to drive them away from the tissue and towards the negative electrode, achieving gentle and efficient antibody removal. Simultaneously, it employs a two-step cascade amplification strategy of small molecule deposition and copper-free click chemical labeling, balancing deep penetration with ultra-high sensitivity. This invention can stably support 6-12 cycles of staining, exhibits high elution efficiency, preserves tissue morphology, and can detect low-abundance targets. It is suitable for spatial analysis of the tumor microenvironment, monitoring of transplant immune rejection, and pathological evaluation of neurodegenerative diseases, demonstrating promising clinical application prospects.

Methods and kits for detecting analytes (such as a plurality of analytes) in a sample utilizing signal amplification (such as DMP-based chemical tag signal amplification) are provided.

The invention belongs to the technical field of biomedicine, and relates to a one-step enzyme chemical detection method for tumor-associated Tn antigens and application of the one-step enzyme chemical detection method, C1GALT1 glycosyl transferase is utilized to realize efficient labeling of Tn antigens in lysate, living cells and biological samples through a one-step enzyme chemical method, fluorescence imaging and Tn enrichment can be carried out, and the Tn antigens in the lysate, the living cells and the biological samples can be efficiently labeled through the C1GALT1 glycosyl transferase. And identification of site-specific Tn protein sites can be carried out through mass spectrometry. Compared with the detection effect of the traditional lectin and the previous two-step enzyme chemical method, the marking efficiency of the method is improved by at least one order of magnitude. The one-step enzyme chemical marker for the Tn antigen can realize multi-dimensional and omnibearing high-specificity, high-sensitivity, stable and rapid detection of the Tn antigen, and has great significance in the aspects of functional research of the Tn antigen in an organism, disease diagnosis and treatment, prognosis, targeted intervention and the like.

The invention belongs to the field of chemical analysis, and particularly relates to a method for quantitatively analyzing complete protein based on a mass spectrum tag. The specific technical scheme comprises the following steps: marking a to-be-detected protein; separating the marked protein by using a size exclusion chromatography-mass spectrometry technology; carrying out quantitative analysis on the protein to be detected by utilizing a mass spectrum mode of combining in-source cracking with multi-reaction monitoring. According to the invention, high-sensitivity absolute quantification of complete protein from top to bottom is realized on low-cost QqQ-MS through a chemical labeling-in-source cleavage-MRM combination strategy. The method for quantitatively analyzing the protein, provided by the invention, is high in universality and good in instrument compatibility.

The invention discloses sampling equipment for lenalidomide quality detection, and relates to the technical field of powder sampling, the sampling equipment comprises an isolation cavity, an outward opening window is arranged in the isolation cavity, a self-adaptive killing mechanism is arranged in the isolation cavity, the outward opening window is arranged on the front surface of the isolation cavity, and the self-adaptive killing mechanism is arranged in the isolation cavity. The self-adaptive killing mechanism comprises a cylindrical cover, a group of external connection seats are fixedly mounted on the outer surface wall of the cylindrical cover, an ultraviolet lamp panel is fixedly connected to the interior of each external connection seat, an annular frame is fixedly mounted at the bottom of the cylindrical cover, the mechanism adopts an operation mode of chemical labeling and physical sterilization, an independent detention space is constructed, and the self-adaptive killing mechanism is arranged in the annular frame. According to the device, the cavity is sealed, isolation from outside air is completed, microorganisms and bacteria contained in the cavity are fully and cleanly treated in a treatment mode of combining fluorescence labeling and ultraviolet killing, the time of a sterile state in the cavity is further prolonged by adopting a physical blocking mode, and it is guaranteed that medicine is prevented from being polluted by the outside air when transferred.

The present invention provides phenylglyoxal-based alkyne (PGA) chemical tags exhibiting high specificity towards protein citrullination sites and other biomolecules containing similarly reactive functional groups. The PGA tags are able to bind to or derivatize biomolecules, such as polypeptides having one or more post-translational modifications (PTMs), such as citrullination. In particular, the PGA tags of the present invention have superior reactivity and selectivity towards ureido groups, and allow for the analysis of biomolecules containing ureido groups facilitated by click chemistry and mass spectrometry (MS) techniques and methods for qualitative and quantitative analysis of biological and clinical samples.

The invention discloses a space-time marking method CAT-Golgi for a living cell Golgi proteome and application of the space-time marking method CAT-Golgi. The space-time labeling method comprises the following steps: sequentially adding a photosensitizer and an aniline probe into living cells to be labeled for incubation, and then irradiating the cells with green light with the wavelength of 520 + / -10nm to realize chemical labeling of a Golgi apparatus proteome, wherein the photosensitizer is a golgi apparatus positioning photosensitizer as shown in a formula I. The method disclosed by the invention can be used for efficiently, simply and conveniently carrying out golgi apparatus proteome capture on various cell lines (HeLa and K562), and compared with a traditional method, the method disclosed by the invention can be used for remarkably improving the subsecretion group specificity (greater than 40%). According to the method, CAT-Golgi proteome analysis can be carried out on cell lines and immune cells which are difficult to transfect.

The application belongs to the field of chemical analysis, and particularly relates to a method for quantitatively analyzing intact proteins based on mass spectrometry tags. The specific technical scheme comprises the following steps: labeling a protein to be measured; separating the labeled protein by using a size exclusion chromatography-mass spectrometry combined technology; and quantitatively analyzing the protein to be measured by using a mass spectrometry mode of in-source fragmentation combined with multiple reaction monitoring. The application realizes the top-down high-sensitivity absolute quantification of intact proteins on a low-cost QqQ-MS by using a chemical labeling-in-source fragmentation-MRM combined strategy. The method for quantitatively analyzing proteins provided by the application has strong universality and good instrument compatibility.

The invention discloses an engineering probiotic-T cell chimera as well as a preparation method and application thereof. Food-grade probiotic lactococcus lactis expressing Flt3L-OX40L double-cytokine fusion protein at the corresponding position of a thallus is successfully constructed in the early stage, then a dibenzocyclooctyne group (DBCO) can be expressed on the surface after chemical labeling, and meanwhile, an azide group (Azido) can be expressed on the surface after a T cell is labeled by glycometabolism. The two can be used for preparing the engineering probiotic-T cell chimera based on bio-orthogonal click chemistry. After successful construction, repeated experiments prove that the engineering probiotic-T cell chimera can promote T cell activation and differentiate into immune memory effector cells (TEM), and the mouse tumor tissue targeting capability of the engineering probiotic-T cell chimera is improved. Meanwhile, animal experiments indicate that the intravenous injection engineering probiotic-T cell chimera has a remarkable effect of inhibiting colorectal cancer tumors.

This invention discloses a chemiluminescent label, a chemiluminescent labeling complex, and their applications, belonging to the field of chemiluminescent immunoassay technology. The chemiluminescent label of this invention is an acridine-pentafluorophenyl ester, prepared by NSP-DMAE-NHS through alkaline hydrolysis, Soxhlet reaction, and substitution reaction. This invention uses this chemiluminescent label for immunoprotein labeling and applies the chemiluminescent labeling complex to a chemiluminescent assay kit. The chemiluminescent label of this invention is not prone to spontaneous hydrolysis and exhibits good stability. In application, it shows low pH requirements for the labeling reaction, achieving good chemical labeling within a fairly wide pH range of 6.0–10.0. The kit prepared by this invention also exhibits higher stability, showing a slow decrease in luminescent activity in accelerated stability studies under high temperature and light irradiation, thus solving the technical problem of insufficient stability encountered by existing acridine esters as chemiluminescent labels.

The present application relates to a kind of high-throughput proteome quantification methods of multi-channel chemical label combined data non-dependence acquisition mode.The method includes:1) prepare 14-plex new chemical label;2) extract the protein of target sample, and enzymatically hydrolyze into polypeptide, and 14-plex tandem mass spectrometry label reagent is used to mark the polypeptide of 14 samples respectively and carries out mixed sample;3) establish the DDA mass spectrum spectrum library of target sample;4) collect the DIA data of polypeptide sample marked with stable 14-plex tandem mass spectrometry label;And 5) using AI algorithm, based on the DIA data collected to identify and quantify target peptide.This application prepares new 14-plex tandem mass spectrometry label, combined with DIA high-throughput proteome quantification method can realize the same peptide segment in different label channel in mass spectrum same window and same scanning being collected, realizes accurate difference quantification of each channel.

Methods of reducing allograft rejection or the risk of allograft rejection in a subject are described. The methods include metabolically labelling an allograft tissue or organ with a chemical tag (such as dinitrophenyl), transplanting the labelled tissue or organ, and administering to the transplant recipient T regulatory (Treg) cells expressing a chimeric antigen receptor (CAR) specific for the chemical tag. The CAR-expressing Treg cells are stimulated upon binding to the chemical tag displayed on the transplanted tissue or organ, leading to activation of the Treg cells and suppression of effector CD8+ T cells, thereby generating of an immunosuppressive tissue environment that reduces the risk of allograft rejection. Kits that include a chemical tag, engineered CAR Treg cells specific for the chemical tag, and / or a viral vector encoding a CAR specific for the chemical tag are also described.

The present invention discloses a non-destructive imaging labeling method and product of αOlig2 and its use, and belongs to the technical field of protein imaging labeling. The present invention labels the fluorescent molecule Cy5 into the structure of αOlig2 by chemical coupling to obtain an antibody protein αOlig2-Cy5 with fluorescence imaging function; utilizes hydrophobic interaction to compound dimerized Gd-DTPA and αOlig2-Cy5 into an antibody probe with fluorescence / magnetic resonance dual imaging function, wherein the introduction of alkyl chains in the molecular structure of dimerized Gd-DTPA increases the binding effect with αOlig2-Cy5. The antibody probe labeling method of the present invention balances the limitations of the weakened target binding ability caused by the change of protein structure in the chemical labeling process and the low image labeling efficiency in the physical compounding process, and retains the target binding ability and image labeling efficiency of the antibody protein to the greatest extent. The antibody labeling method of the present invention is simple and easy to implement, and can be used as a new method for non-destructive labeling of antibody proteins.

To provide a method for identifying polyester resin compositions using chemical tags, identified polyester raw materials, and a method for manufacturing polyester resin products.SOLUTION: There is provided a method for identifying recycled polyester resin that requires the following condition (a). (a) The chemical tag possesses seven or more conjugated double bonds.SELECTED DRAWING: None

The application discloses a chemical labeling system HIM-IP for detecting cell-cell interaction. The system is constructed by modifying tyrosinase and phenol on the surfaces of two cells to be detected respectively, constructing a trapper cell and a prey cell, and then using biotin or single-stranded DNA with a nucleophilic group as a molecular tag for labeling. The system can not only be detected by traditional flow cytometry, but also be compatible with commercial single-cell transcriptome sequencing platforms, realizing the correlation of cell-cell interaction information and transcriptome information at the single-cell level. The system does not need complex genetic engineering modification of cells, the whole modification and labeling process is simple and efficient, and the detection sensitivity of cell-cell interaction is high. By integrating single-cell sequencing technology, the biological mechanism of cell-cell interaction regulation can be further excavated.