48 results about "CD63" patented technology

The application discloses a separation-free plasma tumor-derived extracellular vesicle detection method, relates to the technical field of biology, and comprises CD63, HER2, GPC1 or / and EpCAM; probe sequences are shown in SEQ ID NO.1-SEQ ID NO.4; the probe is applied to the extracellular vesicle detection reagent in the separation-free plasma, single EVs imaging analysis, detection of the image of a single EV after direct dilution of plasma, dynamic immune detection, and statistics of the number of specifically combined EVs. The application applies the separation-free extracellular vesicle detection probe to the immune detection reagent, avoids the separation process of plasma EVs, does not need to use fluorescence detection, can characterize the single EVs binding kinetics, and can detect 1 target EVs in 350 background particles.

The invention discloses an amniotic epithelial cell-derived exosome for treating gestational hypertension and application thereof, and belongs to the technical field of biological medicines. The exosome (hAECs-exos) is obtained by secreting and extracting human amniotic epithelial cells (hAECs), the particle size distribution of the exosome (hAECs-exos) is 30-150 nm, and the exosome (hAECs-exos) expresses exosome specific markers CD9, CD63 and CD81. The invention also provides a preparation method of the exosome. The preparation method comprises the following steps: isolated culture of hAECs, and ultracentrifugal extraction and identification of the exosome. Animal experiments prove that when the exosome is applied to a gestational hypertension disease model animal in an intravenous injection mode, the blood pressure and urine protein level of the gestational hypertension disease model animal can be remarkably reduced, and the fetal rat dysplasia condition is improved. The invention provides a brand-new and effective biological preparation candidate scheme for the treatment of the gestational hypertension disease.

Disclosed are a specific marker combination of mesenchymal stem cell-derived small extracellular vesicles and the use thereof. The characteristic marker combination comprises aminopeptidase N (CD13), integrin β-1 (CD29), membrane glycoprotein THY-1 (CD90), and markers CD9, CD63 and CD81. By using the characteristic marker combination of mesenchymal stem cell-derived small extracellular vesicles for nano-flow cytometry analysis at the single-vesicle level, mesenchymal stem cell-derived small extracellular vesicles can be identified. On the basis of the provided characteristic marker combination, specific modification and engineering on the basis of small extracellular vesicle membrane proteins can further be realized.

The application provides an engineered exosome loaded with a CD63 transmembrane domain-matrix metalloproteinase fusion protein and a preparation method and application thereof. The engineered exosome is secreted by a genetically engineered mesenchymal stem cell, has a CD63 derived transmembrane domain anchored on the membrane, and has a matrix metalloproteinase (MMP) loaded on the inner side of the membrane. The fusion protein comprises, from N-terminus to C-terminus, a fourth transmembrane domain and a C-terminal cytoplasmic tail of a human CD63 protein, a flexible linker peptide, and a human MMP without a natural signal peptide. The application realizes efficient and directional loading of MMPs by using a natural biological generation path of the exosome, shields the inhibition of MMPs by metalloproteinase tissue inhibitors in a scar tissue by using the membrane of the exosome, realizes specific activation of MMPs in a target cell, degrades abnormally deposited collagen, adjusts the ratio of type I and type III collagens, and plays an anti-fibrosis and anti-scar formation role. The application also provides a preparation method of the engineered exosome and application thereof in preparation of a medicine for preventing or treating pathological scars.

The present application provides a fusion protein comprising an anti-CD3 single-domain antibody, an exosome protein CD63 and an RNA binding protein, and uses thereof. The fusion protein of the present application achieves the effects of treating cancer, immune regulation and activating immune cells through various efficacy experiments.

The application discloses an exosome-based protein delivery system and application thereof. The exosome-based protein delivery system comprises a loading subsystem and / or an unloading subsystem, the polypeptide peptide segment expressed by the nucleotide sequence of the loading subsystem comprises CD63, a connecting peptide, an enterokinase cleavage site and a delivered protein or polypeptide, and the polypeptide peptide segment expressed by the nucleotide sequence of the unloading subsystem comprises CD63, a connecting peptide, enterokinase or enterokinase light chain. The application also constructs a human umbilical cord mesenchymal stem cell strain hUC-MSC CDNF‑exo on November 13, 2024 in the Guangdong Provincial Microbial Culture Collection Center, and the preservation number is GDMCC NO:65478. The human umbilical cord mesenchymal stem cell strain hUC-MSC CDNF‑exo The purpose of delivering CDNF into the cytoplasm of target cells is achieved.

The application discloses a nucleic acid free extraction reagent of plasma exosome and a direct PCR amplification kit. The nucleic acid free extraction reagent comprises 0.001-0.1% polidocanol and 1-100 mM Tris-HCl in mass volume concentration, and the solvent is water. The direct PCR amplification kit provided by the application comprises the nucleic acid free extraction reagent and an immunomagnetic bead suspension, a washing liquid, a free extraction reagent and a PCR Mix solution; the surface of the immunomagnetic bead is simultaneously coated with three exosome specific antibodies of CD9, CD63 and CD81, so that the recovery rate of the exosome can be greatly improved during the extraction and purification of the plasma exosome; the PCR Mix solution contains an anti-inhibitory component, can effectively neutralize the inhibitors such as proteins in the PCR system, and ensures the amplification efficiency of the PCR. The nucleic acid free extraction reagent of the application can realize the amplification detection of the exosome nucleic acid free extraction, and simplifies the operation process of the PCR detection.

The invention belongs to the field of biomedicine, and particularly relates to a bifunctional modified magnetic bead and an extraction method of exosomes in cerebrospinal fluid. Aiming at the defects of an existing magnetic bead capture method in the aspect of removing impure protein, the invention firstly provides a bifunctional modified magnetic bead, a CD63 antibody and a sulphobetaine type zwitterionic polymer are modified on the surface of the bifunctional modified magnetic bead, specific capture of the exosome is realized on the basis of antigen-antibody specific binding of CD63 and the exosome, and meanwhile, the adsorption effect on the impure protein is reduced. The magnetic beads are applied to extraction of exosomes in cerebrospinal fluid, and the technical defects that the sample size of the cerebrospinal fluid is small, and the recovery rate of a traditional method is not high are effectively overcome.

The invention provides methods for prognosing an increased risk of early onset preeclampsia (EOPE) in a pregnant subject before the disease onset. The methods comprises quantifying small extracellular vesicles that express pairs of biomarkers selected from CD10, CD63 and placental alkaline phosphatase (PLAP) in a sample obtained from the subject.

The application discloses a kind of targeted exosome chimeric antigen receptor molecules for treating HIV infection and its application, belong to the field of biological medicine technology.The application is packaged into targeted exosome by coding anti-HIV CAR mRNA directionally.The nano antibody sequence of targeted CD3, CD4, CD8+T receptor on T cell is fused to the N-terminal of LAMP-2B to construct specific targeting plasmid, so that the targeted peptide is expressed on the surface of exosome membrane, the expression plasmid containing RNA binding protein L7A and exosome tag protein CD63 is constructed, and the exosome loaded with CAR mRNA and targeted to T cell is obtained by three-plasmid co-transfection of HEK293F cells containing C / D box CAR plasmid.The exosome can construct human CAR-T cell when incubated with PBMC in vitro or back into humanized mouse in vivo, and has significant killing virus infected cell activity.

This invention broadly relates to the field of bioassay technology, and more specifically, to a method for enriching exosomes and the conjugates used in this method. The conjugates are formed by conjugating a CD63 antibody or its antigen-binding fragment with a matrix support; wherein the heavy chain complementarity-determining regions HCDR1, HCDR2, and HCDR3 of the CD63 antibody or its antigen-binding fragment are shown in SEQ ID NO: 1-3, and the light chain complementarity-determining regions LCDR1, LCDR2, and LCDR3 of the antibody or its antigen-binding fragment are shown in SEQ ID NO: 4-6. The conjugates provided by this invention have high affinity and high purification efficiency, and show great promise for application in exosome purification.

The multifunctional antimonene-gold nano composite material (AM-Au NCs) is rapidly, simply and conveniently prepared and is used for multi-analyte fluorescence sensing (such as metal ions, CD63 and tumor-derived exosomes), molecular information coding and protection. Sodium dodecyl benzene sulfonate, AM, Au < 3 + > and NaBH4 are mixed by using different formulas at room temperature to prepare AM-Au NCs with an adjustable gold nanostructure and characteristics. And interestingness is that AM-Au NCs doped with the gold nanoparticles show wide fluorescence quenching capability on the nanosheets, and the fluorescence quenching capability is remarkably different in different dyes and DNA (deoxyribonucleic acid) consisting of different basic groups. A fluorescent sensing platform constructed by AM-Au NCs is combined with a dye or an aptamer and is used for detecting metal ions, CD63 and tumor-derived exosomes. In real samples, through binary or digital coding of selective response and diversified DNA sequences of sensing systems, a series of molecular encryption and hiding technologies are provided and applied to realize molecular-level protection of special text information.

The application discloses a preparation method and application of an engineered exosome, and relates to the technical field of biology.The application provides a preparation method of an engineered exosome, and a carrier construction step is provided for constructing CD63-VEGF into a FUGW carrier, and replacing an Ubc promoter in the FUGW carrier with EF1a to obtain FUGW(EF1a)-CD63-VEGF, wherein the sequence of the FUGW(EF1a)-CD63-VEGF is shown as SEQ ID NO:5.The application successfully develops the engineered exosome capable of carrying VEGF by using synthetic biology technology, and the application provides a train of thought for exosome drugs and drug development with the exosome as a delivery carrier.The engineered exosome carrying VEGF has a remarkable effect on promoting hair growth.

The invention discloses an extraction method and application of membranous small vesicles. The extraction method comprises the following steps: collecting a biological sample, and selecting at least one of blood, urine and cell culture supernatant; centrifuging the sample at 10,000 xg for 20 minutes, and removing cell debris and apoptotic bodies to obtain a pretreated sample; adding 20% by volume of sample separation liquid and CD63 / CD81 antibody modified magnetic beads into the pretreated sample, and incubating and capturing target vesicles; freezing the captured vesicles with liquid nitrogen for 15-35 minutes, and dissolving the vesicles with a 33-43 DEG C water bath; and cracking the dissolved vesicles with 0.5 W / cm < 2 > ultrasonic waves to finally obtain the intact vesicles of 30-70 nm and soluble protein factors. Original plasma or urine can be directly treated, dilution is not needed, and loss of a target object is avoided; a low-cost and degradable biomimetic polymer is developed to replace an imported kit, and the cost is reduced by 75%; the whole process lt; the time is 2.5 hours, which is superior to that of an ultracentrifugation method; and the extracted membranous small vesicles have good integrity and are suitable for liquid biopsy and drug carrier construction.

The present invention pertains to combination comprising a CD63-binding molecule and a drug and / or a tracer for use in a method of therapy and / or in a method of in-vivo diagnosis, wherein the drug and / or the tracer is delivered to a myofibroblast.

The invention relates to the technical field of biological medicines, and provides a high VEGF protein / miR-210 extracellular vesicle induced by a serum-free combined culture medium and application of the high VEGF protein / miR-210 extracellular vesicle. According to the invention, high VEGF protein / miR-210 extracellular vesicles are obtained based on induction of a serum-free culture medium; the serum-free culture medium comprises a basic culture medium component combination: sugar-free RPMI1640 and MCDB131 in a volume ratio of 4: 1; the metabolic regulation additive is prepared by synergistically adding sodium pyruvate, cysteine and a proline hydroxylase inhibitor and taking alanine glutamine as an energy substitute; the growth factor is a combination of polyvinyl alcohol, 4-hydroxyethylpiperazine ethanesulfonic acid, ITS-X, epidermal growth factors and basic fibroblast growth factors, wherein the ITS-X, the epidermal growth factors and the basic fibroblast growth factors are added before use. CD63 and CD81 are specifically expressed on the surface of the prepared high VEGF protein / miR-210 extracellular vesicle, the high VEGF protein / miR-210 extracellular vesicle is rich in VEGF protein and miR-210, and the high VEGF protein / miR-210 extracellular vesicle is applied to targeted therapy of allergic rhinitis by inhibiting Th2 type immunoreaction.

This invention discloses a high-BNIP3-loaded exosome, its preparation method, and its applications, relating to the field of biomedical engineering technology. The high-BNIP3-loaded exosomes are obtained by pre-treating human bone marrow mesenchymal stem cells transfected with a recombinant expression vector carrying the BNIP3 gene, followed by 24 hours of hypoxia at 1% oxygen concentration. The exosomes have a particle size of 40-130 nm and highly express BNIP3 and exosome markers HSP70, CD63, and TSG101. The preparation method includes constructing a pcDNA3.1-BNIP3 recombinant vector, transfecting and screening stable-expressing engineered cells, hypoxia pre-treatment, and purification by gradient centrifugation combined with ultracentrifugation. This invention achieves efficient loading and targeted delivery of BNIP3 through a synergistic strategy of genetic engineering and microenvironment simulation, significantly improving the endocytosis efficiency of nucleus pulposus cells, activating mitophagy, restoring mitochondrial function, and promoting extracellular matrix synthesis. It can be used to prepare drugs for treating intervertebral disc degeneration and has excellent clinical application prospects.

The invention discloses composite hydrogel as well as a preparation method and application thereof. The preparation method comprises the following steps: immersing a reticular metal substrate into a first pre-polymerization solution, taking out the reticular metal substrate, and performing cross-linking polymerization; immersing into a second pre-polymerization solution, taking out, and carrying out cross-linking polymerization; dissolving and removing the reticular metal substrate to obtain double-network hydrogel; a double-network hydrogel is modified by adopting a DNA long chain to obtain a composite hydrogel, the 5'terminal amino group of the DNA long chain is modified, the DNA long chain is grafted on the double-network hydrogel through amidation reaction, and the DNA long chain comprises a plurality of repeated CD63 aptamer sequences. According to the preparation method, the reticular metal substrate is taken as a template, polymerization is initiated in sequence to form the double-network hydrogel, and an internal regular three-dimensional macroporous structure is ensured, so that exosomes are easy to diffuse into the material; comprising a plurality of repeated CD63 aptamer sequences and carboxyl groups of the double-network hydrogel, so that abundant capture sites are formed, and specific efficient separation and enrichment of exosomes in a sample can be realized.

The application discloses a kind of based on microfluidic and AI image recognition basophil activation detection method and system.The method comprises: the peripheral blood sample to be measured is injected into the microfluidic chip device containing conical microporous filter membrane and enriched basophil;In reaction cavity, pre-mixed dry CD63-FITC and CD203c-PE fluorescently labeled antibody and allergen stimulant are added;After incubation at 37 DEG C, 512×512 pixel high-resolution image is obtained by digital microscopic imaging system;Improved target detection model is used to identify activated cells, the model uses lightweight FasterNet backbone network and dynamic attention mechanism, generates cell activation heat map by Gaussian kernel density estimation, combines fluorescence intensity weighted calculation activation index and outputs visual report.System includes microfluidic chip module, fluorescently labeled module, temperature control incubation module, image acquisition module, AI analysis module and man-machine interaction module.The application realizes high sensitivity, high specificity and automatic detection, and is suitable for clinical diagnosis of allergic diseases and blood diseases.

The invention discloses a biomarker detection kit and a detection method for prognosis evaluation of multiple myeloma, and relates to the technical field of biological medicine detection.According to the kit, circMYC and circSETD2 bicyclic RNA combination in serum exosomes serves as a core prognosis marker, and the kit contains an exosome solid phase capture reagent, sulfydryl modified silicon dioxide magnetic beads and the like; the isothermal amplification detection reagent contains a specific SDA primer pair and the like; a double quality control reagent containing a CD63 peptide fragment marker and the like; positive control and negative control are provided; the serum exosome bicyclic RNA is selected as a core prognostic marker, a complete detection system is constructed, an exclusive reagent efficiently enriches and purifies the exosome, the detection specificity and accuracy are improved, a reliable basis is provided for prognostic layering, integrated operation is achieved, the process is simplified, clinical application is achieved, quantitative analysis is matched with a clear interpretation standard, and the detection efficiency is improved. The reagent is stable and convenient, the result is comparable, and the clinical application value is improved.

The invention provides a targeting exosome for hair regeneration and a preparation method thereof. The method comprises the following steps: separating dermal mastoid cells with a cell marker CD133 from a hair follicle strain of a healthy donor, and culturing the dermal mastoid cells in a laboratory conforming to a Taiwan GTP cell preparation factory domain in China in a heterologous and serum-free environment. Then, a culture solution containing exosomes is collected from the cell culture solution, and the multiple exosomes have surface markers such as CD9, CD63 and CD81; and finally, separating, purifying and concentrating the exosome from the culture solution by utilizing a tangential flow filtration system method or a size exclusion chromatography method and other technologies to obtain the final targeting exosome freeze-dried powder product. The invention also provides a liquid hair growth preparation prepared from the exosome product.

The invention belongs to the technical field of biology, and particularly relates to a cell exosome and a separation method and application of an animal cell exosome based on two-stage filtration and multi-stage centrifugation. The separation method comprises the steps of cell culture, primary centrifugation, secondary centrifugation, two-stage filtration, two times of ultracentrifugation, resuspension identification and the like. Optimized parameters are as follows: inoculating 1 * 10 < 6 >-3 * 10 < 6 > cells, culturing for 48-52 hours by using 10% exosome-deficient serum, carrying out gradient centrifugation of 600-700g and 12000-15000g, carrying out two-stage filtration of 0.45 [mu] m and 0.22 [mu] m, carrying out two times of ultracentrifugation of 110000-12000g, and carrying out PBS (Phosphate Buffer Solution) resuspension. The obtained exosome contains TSG101 and CD63 markers, is high in purity and complete in structure, can be used for preparing tumor diagnosis markers or delivery carriers, and has the advantages of being low in cost, high in universality and capable of being standardized.

The invention relates to a kit for evaluating the risk of PD-1 immune drug-related adverse reactions (irAE). The kit comprises a component for detecting a specific biomarker group, and the biomarker group comprises a protein marker selected from the group consisting of ATP6V1G1, GOT2, CD63 and CHCHD4, and / or a metabolite marker selected from the group consisting of kynurenine, tryptophan and phosphatidylcholine PC (17: 117: 1) and PC (17: 226: 7). According to the kit, by analyzing biomarker signals detected by the components of the kit, objective indexes can be provided for evaluating the risk of irAE of a patient. The kit disclosed by the invention is suitable for detecting a biological sample in an in-vitro environment and provides information support for clinical decision-making.

The invention relates to a medicine for improving expression of lysosome related transmembrane protein 4B and a screening method of the medicine, and belongs to the technical field of medicine screening. According to the invention, a fluorescent marker is accurately inserted into a target protein LAPTM4B gene locus, so that real-time monitoring of dynamic change of endogenous protein is realized; meanwhile, membrane protein CD63 is introduced as an internal reference signal, a high-throughput automatic drug screening and image analysis system is utilized to quantitatively screen compounds in a large-scale drug library, finally, drugs capable of improving LAPTM4B expression are screened out, and a new direction is provided for subsequent research on therapeutic targets related to lysosome-related transmembrane protein 4B. Meanwhile, the screened medicine is good in specificity, LAPTM4B expression can be improved, but CD63 is not affected, so that time-space precise control over LAPTM4B activity can be achieved, and the method has important significance in construction of an in-vitro / in-vivo instantaneous activation model.

The invention relates to the technical field of biological detection, and discloses a saliva exosome detection kit for non-invasive auxiliary screening of Alzheimer's disease and a preparation method of the saliva exosome detection kit. The specific exosome enrichment system composed of the anti-L1CAM / CD63 double antibody coated capture plate and the anti-MUC1 / KRT19Fab fragment blocking agent is constructed for the first time, about 85% of oral cavity source exosomes can be effectively removed, and the capture purity of neuron source exosomes is remarkably improved; the special saliva preserving fluid containing DTT and a specific enzyme inhibitor is developed, the technical bottlenecks that saliva RNA is easy to degrade and a sample is uneven in viscosity are overcome, and the room temperature stability is prolonged to 8 hours; an RT-qPCR detection system taking saliva exosome miRNA-135a as a core and taking internal reference miRNA-16-5p standardization is established, and clinical sample verification shows that the kit can effectively distinguish the Alzheimer's disease patient from a healthy control, shows excellent inter-group distinguishing ability, and provides a stable, efficient, reliable and complete technical scheme for solving the problem of AD non-invasive early screening.

The invention discloses an extracellular vesicle for treating facial nerve injury and a preparation method thereof. The hNSCs-EVs are separated from an hNSCs conditioned culture medium through an ultracentrifugation method, the particle size of the hNSCs-EVs is 100-150nm, and the hNSCs-EVs express CD9, CD63 and TSG101 markers. Experiments prove that when hNSC-EVs is locally injected to the injured part of facial nerves, functional recovery of the facial nerves can be remarkably promoted, electrophysiological signals can be improved, and axon regeneration, myelin sheath formation and Schwann cell activation can be effectively promoted. The invention provides a brand new treatment strategy which is safe, effective, easy to store and standard for facial nerve injury, and has a wide clinical application prospect.

The present application relates to a kind of inhaled allergen basophil activation detection method and its application.Specifically, the present application provides a reagent combination for detection, which includes: a first antibody against human CD203c, a second antibody against human CD123, a third antibody against human CD45, a fourth antibody against human CD63, a positive control, and an inhaled allergen.The method of the present application can quickly and accurately detect basophil activation by blood collection, activation reagent configuration, activation, termination of activation, antibody labeling, red blood cell lysis, flow cytometry detection, result analysis and judgment.The present application can be used for diagnosing inhaled allergen sensitized allergic diseases, identifying inhaled allergen and its components, predicting the severity of allergy and monitoring the efficacy.

This invention belongs to the field of nanobody technology, and relates to an anti-CD63 protein nanobody, its preparation method, and its application. The amino acid sequence of the nanobody is shown in SEQ ID NO.1. This invention utilizes CD63 protein as an immunogen to immunize alpacas, extracts total RNA from their peripheral blood lymphocytes, amplifies the VHH fragment via reverse transcription and two rounds of nested PCR, digests and ligates the VHH fragment with the pComb3XSS plasmid using enzymes, and transforms it into TG1 competent cells. Using phage display technology, after three rounds of screening, an anti-CD63 protein nanobody was successfully screened. This nanobody exhibits high specificity and good binding affinity, providing a new direction for the identification, enrichment, and purification of extracellular vesicles or exosomes, and for disease (such as tumor) diagnosis and targeted drug delivery.

The application discloses an application of perirenal adipose tissue exosomes rich in miRNA-486 in inducing endothelial dysfunction, wherein the perirenal adipose tissue exosomes are derived from adipose tissue around mammalian kidneys, rich in micro miRNA-486, have a particle size of 30-150 nanometers, have surface markers CD63, CD81 and TSG101, and have an expression level of the miRNA-486 in the perirenal adipose tissue exosomes increased by 3-5 times relative to a normal control, so that the perirenal adipose tissue exosomes can transmit the miRNA-486 to cells when being taken by vascular endothelial cells, and induce abnormal functions of the endothelial cells. The application proves that the perirenal adipose tissue exosomes rich in miR-486 can be taken by vascular endothelial cells in a specific manner through in-vivo and in-vitro experiments.

The invention relates to a membrane fusion liposome delivery system and a preparation method and application thereof, and belongs to the technical field of liposome preparation. The invention provides a membrane fusion liposome delivery system. The system takes lipidosome as a carrier, the surface of the lipidosome is modified with cRGD targeting molecules, and the interior of the lipidosome is loaded with a TPP-modified antitumor drug lonidamine; a DNA device CLTD is anchored on a liposome membrane through cholesterol, and the device is formed by a short-chain CAP containing an APE1 restriction enzyme cutting site and a long-chain LTD containing a CD63 aptamer and a RelA aptamer through base complementary pairing. The APE1 enzyme highly expressed in a tumor microenvironment is used as a specific trigger signal, and precise activated release of the drug in tumor cells and space-time controllable degradation of RelA protein are achieved. According to the system, chemical drug inhibition and nucleic acid aptamer mediated protein degradation are synergistically linked, off-target toxicity is effectively overcome, and a new strategy is provided for precise treatment of cancers.