153 results about "Protein labeling" patented technology

The present invention provides systems, devices, apparatuses and methods for automated bioprocessing. Examples of protocols and bioprocessing procedures suitable for the present invention include but are not limited to: immunoprecipitation, chromatin immunoprecipitation, recombinant protein isolation, nucleic acid separation and isolation, protein labeling, separation and isolation, cell separation and isolation, food safety analysis and automatic bead based separation. In some embodiments, the invention provides automated systems, automated devices, automated cartridges and automated methods of western blot processing. Other embodiments include automated systems, automated devices, automated cartridges and automated methods for separation, preparation and purification of nucleic acids, such as DNA or RNA or fragments thereof, including plasmid DNA, genomic DNA, bacterial DNA, viral DNA and any other DNA, and for automated systems, automated devices, automated cartridges and automated methods for processing, separation and purification of proteins, peptides and the like.

A method for labeling acyl carrier protein (ACP) fusion proteins with a wide variety of different labels is disclosed. The method relies on the transfer of a label from a coenzyme A type substrate to an ACP fusion protein using a holo-acyl carrier protein synthase (ACPS) or a homologue thereof. The method allows detecting and manipulating the fusion protein, both in vitro and in vivo, by attaching molecules to the fusion proteins that introduce a new physical or chemical property to the fusion protein. Examples of such labels are, among others, spectroscopic probes or reporter molecules, affinity tags, molecules generating reactive radicals, cross-linkers, ligands mediating protein-protein interactions or molecules suitable for the immobilization of the fusion protein.

The invention provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ biotin ligase and biotin analogs.

The invention provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ biotin ligase mutants and biotin analogs recognized by such mutants.

An improved method for protein labeling comprising the steps of providing a synthetic small molecule tag, providing a target protein to be tagged, providing at least two enzymes for catalyzing a conjugation reaction between the tag and the target protein, incubating the tag, the protein and the enzyme, and allowing the tag to conjugate to the target protein. The tag may embody at least one structural feature of an ubiquitin C-terminus, and the structural feature may comprise a recognition sequence that is recognizable by an ubiquitin activating enzyme.

A method for labeling acyl carrier protein (ACP) fusion proteins with a wide variety of different labels is disclosed. The method relies on the transfer of a label from a coenzyme A type substrate to an ACP fusion protein using a holo-acyl carrier protein synthase (ACPS) or a homologue thereof. The method allows detecting and manipulating the fusion protein, both in vitro and in vivo, by attaching molecules to the fusion proteins that introduce a new physical or chemical property to the fusion protein. Examples of such labels are, among others, spectroscopic probes or reporter molecules, affinity tags, molecules generating reactive radicals, cross-linkers, ligands mediating protein-protein interactions or molecules suitable for the immobilization of the fusion protein.

The invention provides a protein labeling and interaction detection system based on engineered fragments of fluorescent and chromophoric proteins that require fused interacting polypeptides to drive the association of the fragments, and further are soluble and stable, and do not change the solubility of polypeptides to which they are fused. In one embodiment, a test protein X is fused to a sixteen amino acid fragment of GFP (β-strand 10, amino acids 198-214), engineered to not perturb fusion protein solubility. A second test protein Y is fused to a sixteen amino acid fragment of GFP (β-strand 11, amino acids 215-230), engineered to not perturb fusion protein solubility. When X and Y interact, they bring the GFP strands into proximity, and are detected by complementation with a third GFP fragment consisting of GFP amino acids 1-198 (strands 1-9). When GFP strands 10 and 11 are held together by interaction of protein X and Y, they spontaneous association with GFP strands 1-9, resulting in structural complementation, folding, and concomitant GFP fluorescence.

The invention discloses a small-molecular fluorescent probe for SNAP protein labeling as well as a synthesis method and an application of the small-molecular fluorescent probe. The probe molecule synthesis route is simple, the reaction conditions are mild, aftertreatment is simple and convenient, and the probe has specific protein recognition capability and higher response speed. In a water solution, the probe has slight fluorescence signals, is specifically bound with SNAP protein and has the fluorescence enhanced by about 30 times, and the emission wavelength of the probe has blue shift about 530 nm after binding. Compared with existing SNAP fluorescent probes, the probe has high selectivity, can specifically recognize SNAP in a biosystem in the complicated environment and has very important application value in the biological and medical fields.

The invention discloses an albumen microsphere conjugate for detecting acrosin activity and a preparation method and application of the albumen microsphere conjugate. The albumen microsphere conjugate is characterized in that monodisperse micron microspheres obtained by combining fluorescein and the acrosome reaction substrate, namely zona pellucida protein are prepared, the acrosin activity is detected by aid of change in fluorescence intensity, and the specific steps include preparation of monodisperse microspheres modified through surface carboxylation or amination, surface modificaction, fluorescence labeling of protein polypeptide, conjugation of fluorescent protein polypeptide and functional microspheres and detection of the acrosin activity. The albumen microsphere conjugate has the advantages that protease digestion on the microspheres is realized through the polymer microsphere technology and the protein labeling and conjugating technology, and enzymatic activity is evaluated; the albumen microsphere conjugate is mainly applied to flow detection of the acrosin activity, synchronous detection of an acrosome reaction and the acrosin activity can be realized, and acrosome functions can be evaluated more truthfully and accurately.

The invention discloses a yellow fluorescent dye, a method for synthesizing the same and application. The fluorescent dye has a structure shown as the general formula (I), wherein R1 and R2 are selected from chlorine, fluorine, bromine or hydrogen; and n is a positive integer between 3 and 7. The fluorescence emission spectrum of the yellow fluorescent dye is positioned in a yellow light region of visible spectrum; and the yellow fluorescent dye has high fluorescence quantum yield and good fluorescence labeling capacity, and can be used for protein labeling.

The invention relates to a royal jelly freshness detection test paper product and a preparation and application method thereof, which belong to the medicine, foodstuff and biotechnological field. As proved by proteomic technology recently, glucose oxidases, royal jelly proteins 4 and royal jelly proteins 5 in royal jelly can be taken as freshness protein labelings. The invention realizes detection of royal jelly quality with quick speed, low cost and high sensitivity through preparation of a nano colloidal gold detection test paper for detecting the proteins. The detection test paper can be used in the aspects like royal jelly raw material quality identification, product quality control and activity value determination during manufacturing, marketing and using processes, and also can be used for detecting royal jelly content in other relative products. Over 90 percent royal jelly in the world is made in China, and annual output value of the royal jelly is over 2 billion; moreover, with improvement of living standard of people, the royal jelly gradually becomes an important natural health product, thereby development of quick and easy variety detection methods is imperative.

The invention relates to an aminoacyl-tRNA (transfer ribonucleic acid) synthetase mutant containing an amino acid sequence selected from groups consisting of amino acids shown by SEQ ID NO:4 and conservative variants thereof. The invention provides a CpK (short for N<epsilon>-(1-methylcyclopropyl-2-acrylamide)-lysine) translation system for fixed point specific insertion of N<epsilon>-(1-methylcyclopropyl-2-acrylamide)-lysine in a target protein through pairing of orthogonal tRNA and orthogonal aminoacyl-tRNA synthetase, and a method for fixed point specific insertion of CpK in the target protein by using the translation system. The CpK translation system comprises (i) N<epsilon>-(1-methylcyclopropyl-2-acrylamide)-lysine, (ii) the orthogonal aminoacyl-tRNA synthetase, (iii) the orthogonal tRNA and (iv) a nucleic acid for coding the target protein, wherein the orthogonal aminoacyl-tRNA synthetase is used for realizing aminoacylation of the orthogonal tRNA preferentially by using CpK, and the nucleic acid contains at least one selective codon specifically identified by the orthogonal tRNA. The invention also relates to an application of a mutant protein for fixed point specific insertion of CpK in protein labeling through a light click reaction with a tetrazole compound.

A pentamethine cyanine fluorescent dye with N-substituting at β-position of conjugated chain is disclosed. The dye reacts with biosubstrate including protein, liposome and DNA, and shows excellent selectivity. It has good living cell membrane permeability and shows good selectivity in living cell staining In addition, the dye has good stability and low background fluorescence, and can be used together with other fluorescent dyes having consistent wavelengths for single-channel excitation and multi-channel detection. The dye can be used in the fields of protein labeling and detecting, fluorescence immunoassay and living cell selective imaging and the like.

The invention discloses a discrimination and detection test strip for bovine viral diarrhoea virus (BVDV) and a preparation method thereof. A BVDV anti-body is quickly and sensitively detected. The test strip is prepared from a support bottom plate, a coated adsorption material layer, a protein-marked bearing material layer, a sample buffer material layer, a liquid absorption material layer, and an outer layer protection material, wherein the coated adsorption material layer is provided with a test line and a quality control line; the protein labeling of the test strip adopts colloidal gold labeling; the protein used by the protein marking is a phage monoclone which is screened from a phage peptide library and is combined with the BVDV antibody; a peptide epitope expressed by the phage monoclone is LTPHKHHKHLHA. The preparation method of the discrimination and detection test strip comprises the following steps: preparing the gold-labeled phage monoclone, preparation of a gold-labeled pad, preparation of nitrocellulose membranes, and assembly of all parts so as to obtain a final product. According to the discrimination and detection test strip disclosed by the invention, a detectionresult can be obtained in 10 minutes, and the test strip is quick and sensitive, and is easy to operate, and is easily popularized and used in basic level in production and scientific research.

The invention discloses a quantum dot microsphere immunochromatography test strip for detecting the total amount of a novel coronavirus IgM-IgG antibody. The test strip comprises a bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad, wherein the sample pad, the combination pad, the nitrocellulose membrane and the water absorption pad are sequentiallyconnected end to end and adhered to the bottom plate, the combination pad is coated with an N protein-marker and an S protein-marker, and the nitrocellulose membrane is provided with a detection linecoated with an N protein and S protein mixture and a quality control line coated with an N protein antibody and S protein antibody mixture. The total amount of the novel coronavirus antibody is quantitatively detected by utilizing the test strip, the detection sensitivity is high, the accuracy is good, and the repeatability and the stability are good; the test strip is simple to operate and low in cost, can quickly and quantitatively detect the total amount level of the SARS-CoV-2 IgM-IgG antibody in a human body, and provides powerful support for epidemic situations.

The invention discloses a reagent for auxiliarily diagnosing lung cancer lymph node metastasis, comprising 8 antibodies which are used for detecting 8 protein markers which are MMP1 (matrix metalloproteinase-1), TIMP1 (tissue inhibitor of metalloproteinases metallopeptidase inhibitor 1), IQGAP1 (IQ motif containing GTPase activating protein 1), TPX2 (targeting protein for Xklp2), uPA (Urokinase-type plasminogen activator), Cathepsin-D, Fascin and pIgR / SC (polymeric immunoglobulin receptor / secretory component). By adopting the 8 antibodies and an immunohistochemical staining result, lung cancer lymph node metastasis can be auxiliarily diagnosed, and the reagent is expected to be used for the risk estimation of lung squamous cell cancer lymph node metastasis and the prognostic prediction. The reagent has high creditability, strong practicability and clinical use value based on the clinical routine immunohistochemical staining technology when the reagent is used for auxiliary diagnosis.

The invention discloses a beta-caryophyllene derivative as well as a preparation method and application thereof. The structural general formula of the beta-caryophyllene derivative provided by the invention is shown by a formula I, wherein R is selected from any one of the following groups: a hydroxy group, an amino group, an ester group and an amide group. The inventor of the invention has verified through experiments that a reaction factor has good reactivity, stability, easily available raw materials and a certain reaction rate at the same time. The application of this reaction factor is proved in corresponding protein labeling and cell labeling experiments, which provides a new idea for the development of related biorthogonal reaction factors.

The invention provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ lipoic acid ligase or mutants thereof, and lipoic acid analogs recognized by lipoic acid ligase and lipoic acid ligase mutants.

The invention discloses a preparation method of goat mycoplasma pneumonia immune colloidal gold labeled rapid diagnosis test paper, which comprises the following steps: preparing a goat mycoplasma pneumonia specific monoclonal antibody by using goat mycoplasma pneumonia BNCC126187 strain RPS11 gene plasmid recombinant purified antigen as an immune antigen; through colloidal gold particle monoclonal antibody protein labeling, purification, quality identification and condition optimization, establishing a mycoplasma pneumonia colloidal gold immunochromatographic assay method; and combining the obtained mouse RPS11 monoclonal antibody and a colloidal gold probe on a glass fiber membrane, and assembling with a solid-phase NC membrane to obtain the gold-labeled detection test strip. The gold-labeled diagnosis and detection test strip developed by the invention has good characteristics of rapidness, sensitivity, specificity and simplicity and convenience in operation on diagnosis and detection of mycoplasma pneumonia of sheep, and lays a good technical foundation for early diagnosis and detection of mycoplasma pneumonia of sheep and development of epidemiological investigation work.

The invention provides a characteristic protein fragment composition for detecting thalassemia and a mass spectrometry model for evaluating therapeutic effect of thalassemia medicine or evaluating effect of a treatment method, wherein the sequence of the characteristic protein fragment composition is shown in SEQ ID No.1-3, respectively. The characteristic protein fragment composition or mass spectrometry model provided by the invention can be used for the diagnosis and screening of thalassemia, as well as the evaluation of the treatment method and drug efficacy of thalassemia. The method is simple and easy to operate, and high in accuracy, and provides new methods and ideas for the evaluation of diagnosis and screening, treatment methods and drug efficacy of thalassemia.

The invention belongs to the field of biomedicines, and concretely relates to amyloid protein-labeled fluorescence probes, and a making method and an application thereof. The fluorescence probes are compounds represented by formula I, and R1 and R2 in the formula I are independently selected from -CF3 and -CN. The invention also relates to the application of the compounds of formula I in amyloid protein labeling as the fluorescence probes. The compounds of formula I can be used for preparing products for diagnosing the Alzheimer's disease. BODIPY fluorescence probe molecules containing a bioactive 1,8-naphthyridyl group as a parent compound are synthesized for the first time, can be applied to beta-amyloid protein labeling, and can be used for the early stage diagnosis of the Alzheimer's disease.

The invention relates to a new fluorescence probe used for protein labeling and a synthetic method thereof. The compound is prepared by the condensation of two water soluble indole compounds, by reaction of an iodoacetamide group of the compounds and mercapto groups of proteins, fixed-point fluorescent labeling of the proteins can be realized. The compound has the advantages of good water solubility, high molar absorptivity, longer excitation wavelengths, easy synthesis, mild protein labeling conditions, and the like. The probe can be used for fixed-point fluorescent labeling of the proteins, and has a great help for the study on protein structure, function and dynamics characteristics. Excitation emission wavelength of the probe is in the range of 500-600nm, the probe has a short linking group, can be used for more accurate description on the protein structure, dynamics characteristics and biological functions.

The invention provides a self-flickering fluorescent dye for super-resolution fluorescence imaging as well as synthesis and application thereof. According to the dye, azetidine is introduced into the4,5-sites of silicon rhodamine to inhibit intramolecular rotation, so that the characteristics of high quantum yield and high stability are achieved; meanwhile, under normal physiological conditions,fluorescence and non-fluorescence forms of the probe can be mutually converted through intramolecular interaction, so that the probe can be applied to living cell super-resolution imaging, the structural formula of the probe is shown as (1), and the dye can be combined with an antibody to perform specific labeling imaging on tubulin in cells. The dye can be widely applied to the fields of proteinlabeling, super-resolution fluorescence imaging and the like.

The invention relates to near-infrared fluorescent dye and a preparation method thereof. The novel phenoxazone near-infrared fluorescent dye is quickly and efficiently synthesized by a nitration reaction, a hydrogenous reduction method and an air oxidation method. The preparation method is environmentally friendly, industrialized production difficulty is low, and the profit is high; the near-infrared fluorescent dye involved in the invention has the characteristics of near infrared emission, high quantum yield with the yield reaching up to 27.55%, excellent dyeing property, good solubility and simple synthesis and preparation, can be widely applied to the aspects of bioimaging techniques, fluorescent probe kits, antibody and protein labelings, tumor targeting markers and drug modification and development, has very good application prospects, can also be widely applied to the industries of anti-counterfeiting printing, pesticides, dyes, building and food testing, and the like at the same time, and has relatively high application value.

The invention discloses an organic mass spectrum flow cytometry technology which comprises the following steps: preparation of a protein labeled flow cytometry probe and a cell labeling method, construction of an organic mass spectrum flow cytometry device, mass spectrum flow cytometry detection of a cell suspension sample, and extraction and analysis of protein labels and metabolite information in a single cell mass spectrum. The technology is high in universality, the device is simple, and rapid, efficient, high-throughput and high-sensitivity sample analysis can be achieved. The flow mass spectrometry probe marks a large number of organic tags on target proteins, so that the detection sensitivity reaches the single cell level, qualitative and quantitative analysis of multiple target proteins and a large number of metabolites at the single cell level can be realized, and a large number of single cell level information is obtained. The technology has wide application prospects in the fields of cell identification, cell typing, research on characteristic difference substances in cells, tumor diagnosis, systematic biology research and the like.

The present disclosure provides compositions and methods of use thereof for labeling peptide and proteins in vitro or in vivo. The methods described herein employ lipoic acid ligase or mutants thereof, and lipoic acid analogs (e.g., lipoic acid analogs comprising a resorufin moiety) recognized by lipoic acid ligase and lipoic acid ligase mutants. Also provided herein is a method of imaging protein-protein interaction via a reaction mediated by lipoic acid ligase.

The invention provides a method for determining a sample source. The method comprises the following steps: (1) carrying out protein content determination treatment on a sample so as to obtain the tumor protein marker content of the sample; (2) standardizing the sample data, wherein the data comprises the tumor protein marker content obtained in the step (1) and the dummy variable conversion data of clinical data; and (3) based on the standardized sample data obtained in the step (2) and a prediction model, determining the probability that the to-be-detected sample comes from a cancer patient.According to the method provided by an embodiment of the invention, whether the to-be-detected sample comes from a cancer patient or not can be accurately and conveniently determined through the prediction model based on the protein content and clinical data of the to-be-detected sample, so a foundation is laid for further research of cancer-related pathogenic mechanisms or early screening of cancer.