79 results about "CD63" patented technology

A method for the measurement of the activation of the basophils following stimulation with an allergen to determine hypersensitivity to some substances, in which a blood sample with an addition of interleukin-3 in a quantity of 0.05 to 50 ng/ml based on the volume of the sample, and of appropriately diluted allergen in a quantity of 0.5 to 100 units/ml is incubated at the temperature corresponding to the physiological environment for 15 to 45 minutes, whereafter a staining with anti-CD63.antibody in an amount of 3 to 30 .mu.l/100 .mu.l of blood and with the antibody against a receptor of the basophil in an amount of 3 to 30 .mu.l/100 .mu.l of blood are added at a temperature of 0 to +10.degree. C. and the sample, after vortexing, is then incubated in an ice bath for 15 to 30 minutes, and then is lysed and subjected to flow cytometry.

The invention relates to the technical field of organism detection and particularly relates to a high-specificity exosome separation, detection and enrichment method which comprises the following steps of constructing a nucleic acid amplification reaction which can be triggered only when phospholipid bilayer and corresponding membrane protein on an exosome membrane exist simultaneously, realizinghigh-specificity exosome detection, and enriching the exosome by virtue of magnetic beads. The method is advantaged in that 1, for a traditional exosome detection method based on the aptamer or the antigen-antibody, in the practical application process, an exosome detection kit is possibly triggered by free exosome membrane proteins such as CD63 protein and CD9 protein in the external environment,a problem that the finally detected signal cannot reflect the quantity of exosomes completely is solved; and 2, the specificity of exosome detection and separation is remarkably improved, interference of free exosome membrane protein in an external environment on exosome detection is reduced, and meanwhile, the higher sensitivity of exosome detection is ensured,

The invention provides a method for separating exosomes. The method at least includes steps of (1), leading CD63 with labels into target cells and culturing the target cells; (2), centrifuging the target cells and then acquiring supernatant; (3), adding magnetic substances into the supernatant and incubating the supernatant and the magnetic substances; (4), arranging the supernatant in magnetic fields; (5), removing the supernatant, adding buffer solution and separation columns into the magnetic substances, eluting the exosomes by the aid of the buffer solution and collecting the exosomes. The magnetic substances contain markers capable of being specifically bound with the labels on the CD63. The exosomes which are about to be eluted are detained on the separation columns. The method for collecting the exosomes has the advantages of high collection efficiency.

The invention relates to the technical field of medical bioengineering, and provided with a specificity surface molecule marker CD63 of a hepatic stem cell. The invention further provided with application of the specificity surface molecule marker CD63 of the hepatic stem cell. A method that the molecule marker is used for quickly separating, culturing and amplifying the hepatic stem cell from a liver specifically comprises the steps that the specificity surface molecule marker CD63 of the hepatic stem cell is used to quickly separate the hepatic stem cell from prepared liver cell suspension through a flow cytometer. The hepatic stem cell of the CD63+ has the characters of self-renewing and bilateral differentiation out of a body, and has therapeutical effects on a damaged mouse liver. According to the specificity surface molecule marker CD63 of the hepatic stem cell and the application of the specificity surface molecule marker CD63 of the hepatic stem cell, the hepatic stem cell obtained by separation can be served as a seed cell for liver drug screening, systematization projection liver and hepatic disease cell therapy, and provides an ideal cell model for liver development and the cell biological property research of the hepatic stem cell.

The invention belongs to the field of medicine detection, and relates to a method for determining an allergen through directly monitoring mast cell degranulation. The method is characterized by comprising the following steps: step 1: culturing an RBL-2H3 cell transfected with CD63-GFP (green fluorescent protein) plasmid and the allergen together; step 2: scanning and collecting the image of the culture by a microscope; and step 3: determining the existence of the allergen according to the existence and the movement of fluorescent particles displayed in the image.

This invention relates to the diagnosis and treatment of cancerous diseases, particularly to the mediation of cytotoxicity of tumor cells; and most particularly to the use of cancerous disease modifying antibodies (CDMAB), optionally in combination with one or more chemotherapeutic agents, as a means for initiating the cytotoxic response. The invention further relates to binding assays which utilize the CDMAB of the instant invention.

The invention discloses a preparation method of a bifunctional hybrid film for tumor exosome self-calibration detection. A bifunctional hybrid film, namely aptamer-BPNSs/Fc/ZIF-67/ITO, is constructedon the basis of simple self-assembly of black phosphorus nanosheets BPNSs, an aptamer and a ferrocene Fc doped cobalt-based metal organic framework ZIF-67 compound on an indium tin oxide ITO electrode. The aptamer marked by methylene blue MB is specifically combined with the CD63 protein to capture protein accurately. The protein is a specific biomolecule carried by breast cancer MCF-7 cell exosome, and realizes detection for the tumor cell exosome. MB is used as a response signal, Fc is used as a reference, and a self-calibration sensor for quantitative detection of the tumor exosome is constructed. Compared with the prior art, the exosome self-calibration detection method is convenient to operate, high in sensitivity, low in cost and high in specificity, can be used as a novel exosome self-calibration detection method, and is used for quantitative detection for exosomes in biomedical samples.

The invention provides a basophilic granulocyte activation and degranulation identification method. The basophilic granulocyte activation and degranulation identification method comprises the following steps that test samples are numbered in sequence, and flow type sample feeding pipes required by testing are marked; a required anti-human CD123, anti-human CCR3, anti-human HLA-DR, anti-human CD203c and/or anti-human CD63 fluorescence labeling flow type antibody combination is added into each flow type sample feeding pipe; mixed whole blood is added into the marked flow type pipes, and shading incubation is conducted at the indoor temperature; red blood cell lysate is added into each sample pipe, and shading incubation is conducted at the indoor temperature again; supernatant is removed in a centrifugal mode after incubation; detection is conducted by means of a flow cytometry, and the number of basophilic granulocytes and the average fluorescence intensity are analyzed. According to the basophilic granulocyte activation and degranulation identification method, 80%-100% of the basophilic granulocytes in peripheral blood can be distinguished from other types of cells under the condition that the basophilic granulocytes do not need to be separated or purified, the basophilic granulocyte activation and/or degranulation state can be identified, whole blood of no more than 100 microlitres is needed by each sample to be tested, and the repeatability is good.

The invention relates to the technical field of ELISA detection kits, and particularly discloses a high-capacity asphalt/epoxy resin-based hard carbon negative electrode material and a preparation method of same. The kit includes a box body, in which two ELISA plates coated with anti-CD63 cloning antibody, an ELISA plate coated with anti-CA125 cloning antibody, 10 ml of HPR-labeled CA125 antibody,10 ml of HPR-labeled CD63 antibody, 20 ml of H2O2 being a color developing agent A solution, 20 ml of TMB being a color developing agent B solution, and two bottles of 5 ml of stop solution. Comparedwith the prior art, by means of the exosome ELISA method, an antigen of the ovarian cancer is quickly detected, thus completing early diagnosis on the ovarian cancer. The kit is high in detection accuracy and specificity and is extensive in detectable sample range, which may include serum, ascites, blood plasma, tissue liquid and the like. The kit is short in reaction period and has good effect.

The invention discloses an extracellular vesicle surface protein specific aptamer screening technology based on magnetic-activated cell sorting. The extracellular vesicle surface protein specific aptamer screening technology includes the following steps of: 1) synthesizing upstream primers, downstream primers and a random library; 2) culturing target cells and obtaining a cell culture supernatant;3) using a grain diameter selection method for extracting extracellular vesicles; 4) using immunomagnetic beads of which the surfaces are modified with CD63 antibodies for capturing the extracellularvesicles; 5) conducting positive screening to acquire library sequences capable of being combined with target extracellular vesicles; 6) conducting positive screening and negative screening alternatively to acquire specific target nucleic acid aptamers; and 7) adopting a flow cytometry technology to detect the enrichment condition of the library. According to the extracellular vesicle surface protein specific aptamer screening technology based on magnetic-activated cell sorting, the grain diameter selection method is adopted to extract the extracellular vesicles, the extracellular vesicles are extracted easily and rapidly, and the recovery rate of the extracellular vesicles is high; the immunomagnetic beads of which the surfaces are modified with the CD63 antibodies are used for capturingexosomes, and the extracellular vesicles can be separated rapidly; and furthermore, positive screening and negative screening are conducted alternatively, thus, the screening efficiency of the targetnucleic acid aptamers can be improved, and by utilizing the screened aptamers, disease liquid biopsies and treatment which take extracellular vesicle surface proteins as targets can be conducted.

The invention relates to the technical field of biosensors, and particularly relates to a biosensor for detecting exosome based on a nucleic acid aptamer. The biosensor comprises an aptamer PTK-7 Apt,CD63 Apt, a connector, a hairpin probe H1, a hairpin probe H2 and a CCRF-CEM. The invention also relates to a production method and an application thereof. Based on specific recognition of the nucleic acid aptamer and a target object, through connection of the connector, two chains of PTK-7 Apt and CD63 Apt are adjacent to each other to form a trigger, and the trigger further opens H1 and the H1opens H2 to trigger an HCR reaction so that amplification of chemiluminescence intensity signals is realized, and the aptamer biosensor is constructed. The sensor has advantages of a high detection speed, simplicity in operation, a low price, a low detection limit, high specificity and the like.

The invention discloses a cynoglossus semilaevis derived exosome sandwich ELISA (Enzyme-linked Immuno Sorbent Assay) detection method and a kit and belongs to the technical field of fish biology, particularly the field of application of marine fish exosome markers. According to the method disclosed by the invention, an anti-CD63 antibody serves as an capture antibody, an anti-HSP90 antibody servesas a detection antibody (an enzyme-labelled antibody), and a double-antibody sandwich ELISA method for detecting the cynoglossus semilaevis derived exosome is developed. The invention further provides a kit capable of identifying and quantifying, used for identifying marker proteins of exosome in cynoglossus semilaevis seminal fluid. The kit has important reference significances for identifying exosome of samples derived from other body fluid of fishes as well as other marker proteins. The method disclosed by the invention is high in specificity, high in efficiency and simple and convenient to operate, does not need large instruments or equipment and expensive chemical reagents, and has popularization value.

This invention relates to the diagnosis and treatment of cancerous diseases, particularly to the mediation of cytotoxicity of tumor cells; and most particularly to the use of cancerous disease modifying antibodies (CDMAB), optionally in combination with one or more chemotherapeutic agents, as a means for initiating the cytotoxic response. The invention further relates to binding assays which utilize the CDMABs of the instant invention.

The invention discloses an exosome surface protein and internal miRNA simultaneous detection chip and a detection technology. The preparation method of the chip comprises the following steps: preparing a Y-shaped array chip by adopting a soft photoetching method according to a drawn mask, sequentially introducing a biotinylated gelatin solution and an avidin solution into the chip, and assemblingthe chip layer by layer; and adding avidin-modified nanoparticles to the surface of the assembled chip, and capturing an antibody CD63 to obtain the micro-fluidic chip. The invention also prepares detection antibodies modified by different quantum dots for exosome surface protein detection, and vesicles containing different molecular beacons for exosome internal miRNA detection. The method has theadvantages of low sample consumption, rapidness, simplicity and convenience in detection, good selectivity, high sensitivity, high flux and the like; in addition, the chip can realize simultaneous detection of exosome-related proteins and miRNAs, and can effectively improve the diagnosis accuracy of corresponding diseases.

The invention discloses a peptide - cisplatin conjugate which is prepared by taking a peptide containing 12 amino acids as a carrier and coupling the pipetide with anticancer drug cisplatin, and the invention makes initial research on the pharmacological properties of the peptide - cisplatin conjugate. Cisplatin is widely applied to traditional cancer treatment, but the advantages of great toxic and side effect and the like thereof restrict the application thereof in the treatment. Therefore, to reduce the toxic and side effect of cisplatin is the key to improve the cisplatin medicine. The peptide - cisplatin conjugate obtained by the coupling of peptide and the cisplatin through chemical reaction has small molecular weight when being used for preparing anti-tumor disease medicine, has noantigenicity, does not cause allergic reaction, helps the medicine to display tumor targeting property, takes envoplakin, CD63 and other antigen as the targets, kills tumors with high efficiency, promotes the apoptosis of tumor cells and reduces the toxic and side effect.

The invention discloses a micro-fluidic Raman chip and a method for detecting exosome in blood based on the micro-fluidic Raman chip, and belongs to the technical field of microfluidics. The micro-fluidic Raman chip is composed of four inlets, an outlet, a valve, a mixing chamber and a Raman detection area. Serum samples and CD63 antibody magnetic beads are pumped into the chip through an inlet 1and an inlet 3 respectively, and when two solutions flow through the staggered triangular micro-column mixing chambers, the two solutions are fully mixed and subjected to immunoreaction. And the CD63magnetic bead exosome compound is fixed in the Raman detection area through magnetic force. And the EpCAM functionalized Raman spheres are added from the inlet 4 to be mixed and reacted with the CD63magnetic bead exosome compound, and finall Raman detection is performed. The chip can be used for rapidly, simply, sensitively and quantitatively detecting the exosome in a serum sample, has strong detection specificity, is simple to operate, and is beneficial to more accurately evaluating the diagnosis of various diseases and guiding the treatment.

The invention belongs to the technical field of poultry disease detection, and particularly relates to a patent application of an ELISA (enzyme-linked immuno sorbent assay) detection method for poultry-derived exosomes. The ELISA detection method of the poultry-derived exosome comprises the steps of sample preparation, coating, sealing, sample adding, detection, result judgment and the like. In the prior art, human-derived exosome surface marker proteins CD9 and CD81 are taken as targets in an exosome-based ELISA detection method, but the marker proteins are not suitable for poultry, namely, the marker proteins are not suitable for detecting poultry-derived exosomes. Therefore, the poultry-derived exosome surface marker protein CD63 is used as a marker, poultry-derived CD63 protein is prepared, corresponding multi-antibody serum is prepared, and on the basis, an ELISA detection method of the poultry-derived exosome is further designed and developed, so that a certain technical foundation can be laid for poultry-related physiological work research.

The invention belongs to the technical field of cell secretion extraction, and discloses a method for extracting exosome derived from human hair follicle dermal papilla cells (DPC-Exos). The method isas below: identifying DPC-Exos by controlled resistance pulse sensing (TRPS) analysis, electron microscopy and Western Blot; and adding the DPC-Exos to a culture medium of human hair follicle outer root sheath cells (ORSCs) cultured in vitro. The DPC-Exos are identified to be about 105 nm in diameter and expressing tumor susceptibility gene (TSG) 101, differentiation cluster (CD) 9 and CD63; DPC-Exos promote proliferation and migration of human ORSCs, and mRNA and protein levels of beta-catenin and Sonichedgehog (Shh) in ORSCs are detected to be up-regulated after treatment; transdermal injection of DPC-Exos accelerates an HF growth phase of mice and delays the occurrence of regression. Immunohistochemical analysis shows that the expression levels of beta-catenin and Shh are up-regulatedin the hair follicles of an experimental group. DPC-Exos help regulate the growth and growth cycle of the HF and provide a new potential for the treatment of clinical hair loss.

The invention provides a DNA molecular machine for detecting exosome, which is formed by combining a characteristic Substate chain and an Aptamer locked Motor chain on the surface of gold nanoparticles (GNP), and is used for fluorescence detection of a specific biomarker in the exosome so as to determine the concentration of the exosome. According to the invention, CD63 protein on an exosome membrane and CD63 aptamer for locking a Motor chain on a DNA molecular machine are used for specific recognition, thus unlocking a Motor chain, combining the Motor chain with a Substate chain, and cuttingoff the Substate chain by the restriction endonuclease, so that the DNA molecular machine begins to operate and releases fluorescent molecules. The DNA molecular machine provided by the invention hashigh specificity and sensitivity and is low in cost; and the CD63 aptamer can also be replaced with aptamers of other specific biomarkers, so that the detection range of a DNA molecular machine is expanded.

The invention provides a perch immunity-related SNP site and application thereof in breeding. By comparing CD63 genes of different families, the SNP site related to disease resistance is obtained, anda screening marker is provided for screening disease-resistant perches. The invention firstly provides the perch immunity-related SNP site, the SNP site is located at the 223 site of the perch CD63 gene with the nucleotide sequence of SEQ ID NO: 1, and the basic group of the SNP site is C or A. The invention further provides a PCR amplification or sequencing primer for detecting the SNP site. The perch immunity-related SNP site is obtained through screening, perch individuals with higher resistance to pathogenic bacteria can be screened out by detecting the SNP site, and therefore the genetic breeding process of perches is accelerated.

The invention relates to the technical field of biology, and discloses a reagent for labeling an exosome and a preparation method and an application method thereof. The reagent is composed of a mercapto modified CD63 aptamer and gold nano particles. The mole ratio of the aptamer to gold nano particles is 50-100:1. According to the preparation method, the mercapto modified CD63 aptamer is used to modify gold nano particles to obtain the reagent, and the mole ratio of the aptamer to gold nano particles is 50-100:1. According to the application method, at a room temperature, the reagent is mixedwith an exosome to be labeled, the mixture is stirred for 2 hours, and the volume ratio of the reagent to the exosome is 1:20-50. The reagent can label an exosome, moreover the price of the mercapto modified CD63 aptamer is low, the experiment cost is reduced effectively, and furthermore, the aptamer is easy to store and is resistant to deterioration.