98 results about "Cell-cell fusion" patented technology

Disclosed herein include methods, compositions, and kits suitable for use in e.g., nucleic acid delivery. Provided are compositions (e.g., nucleic acid compositions) comprising polynucleotide(s) encoding a fusion protein, a dimerization fusion protein, a soluble RBP, an adapter fusion protein, and / or a cell fusion protein. The compositions also comprise one or more polynucleotides comprising one or more RNA cargo molecules. The fusion protein, dimerization fusion protein, and / or adapter fusion protein can comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD). Recruitment of one or more ESCRT proteins results in secretion of enveloped nanoparticles (ENPs) from a cell in which at least one of a fusion protein, a dimerization fusion protein, and / or an adapter fusion protein are expressed. The ENPs can comprise the one or more RNA cargo molecules. There are also provided populations of ENPs in some embodiments.

The invention relates to a multifunctional integrated organoid culture device. The device comprises a main body, and a liquid input module, a temperature control module and an organoid module which are fixed on the main body, the main body comprises an adjusting assembly and a main control chip which are connected with each other, and the main control chip is connected with the temperature control module and the liquid input module and used for adjusting the flow speed of the liquid input module and the temperature of the temperature control module; the liquid input module is connected with the organ-like culture module and used for conveying cell sap to the organ-like culture module, the organ-like module is provided with a cell anchoring array and used for fixing and culturing cells, and the temperature control module is located between the liquid input module and the organ-like module and used for controlling the temperature of the organ-like culture module. The temperature detection module is used for detecting the temperature of the liquid input module and adjusting the temperature of the organ-like module. Compared with the prior art, the invention has the advantages of high control precision, multi-cell fusion, abundant functions and the like.

The application belongs to the technical field of biology and particularly relates to an African swine fever virus pNP868R protein monoclonal antibody and a preparation method and application thereof. The application takes the pNP868R protein prepared by eukaryotic expression as an antigen, immunizes BALB / c mice, uses hybridoma technology, and obtains a hybridoma cell line capable of stably secreting the anti-African swine fever virus pNP868R protein monoclonal antibody through cell fusion, indirect ELISA method screening, limited dilution method subcloning and hybridoma continuous passage culture. The application also obtains an African swine fever virus pNP868R protein monoclonal antibody. The monoclonal antibody has high affinity with the pNP868R protein and can be applied in Western-Blot experiments, indirect immunofluorescence experiments, immunoprecipitation experiments and the capture of proteins interacting with pNP868R, and can be used for the diagnosis, prevention or treatment of African swine fever.

The invention relates to a hybridoma cell strain secreting a rofecoxib monoclonal antibody and application of the hybridoma cell strain, and belongs to the technical field of immunodetection. According to the invention, a hybridoma cell strain secreting the rofecoxib monoclonal antibody is obtained through screening by preparing a rofecoxib complete antigen and carrying out animal immune reaction and cell fusion steps, the hybridoma cell strain is preserved in China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC NO.46734. The monoclonal antibody secreted by the hybridoma cell strain has good sensitivity and specificity, the IC50 value to rofecoxib is 20.0 ng / mL, the cross reaction rate to various rofecoxib functional analogues is smaller than 1%, the monoclonal antibody can be applied to preparation of rofecoxib detection products, and an efficient detection method and means are provided for detection of rofecoxib residues in food.

The invention discloses a feline leukemia virus p27 protein monoclonal antibody, a hybridoma cell strain and application thereof, and relates to the technical field of biology, the amino acid sequence of a heavy chain variable region of the monoclonal antibody is as shown in SEQ ID NO.1 in a sequence table, and the amino acid sequence of a light chain variable region of the monoclonal antibody is as shown in SEQ ID NO.2 in the sequence table. According to the invention, a cell fusion technology is utilized to establish a hybridoma cell strain p27-3A5 capable of secreting a p27 protein monoclonal antibody, and a specific monoclonal antibody 3A5 is obtained. And the minimum linear epitope identified by the antibody is identified, so that the establishment of subsequent preparation of a diagnostic kit such as a colloidal gold test strip is facilitated.

This invention belongs to the field of immunology and discloses a hybridoma cell line and the preparation and application of its secreted monoclonal antibody that uniformly recognizes tylosin / tilmicosin. The monoclonal antibody hybridoma cell line is named DES-AOAA-14D5, with accession number CGMCC NO: 45303. This invention utilizes decarboxymethyl tylosin and oxycarboxymethyl hydroxylamine to synthesize a hapten, conjugates a carrier protein to prepare a complete antigen, immunizes mice, and, through cell fusion and screening techniques, prepares a hybridoma cell line that can secrete a monoclonal antibody that uniformly recognizes tylosin / tilmicosin. IC50 50 The effective concentrations were 1.59 and 1.72 ng / mL, respectively, with a cross-reactivity rate as high as 92.44%. An immunoassay method for the simultaneous and accurate detection of tylosin / tilmicosin in milk was established using this antibody. The cut-off value for both tylosin and tilmicosin was 16 ng / mL, and no cross-reactivity was observed with other structural analogs. The detection method of this invention shows promising application prospects.

The disclosure provides the structure of an MFSD2A-SYNC2 complex together with functional data that revealed important molecular aspects of MFSD2A transport cycle, receptor-mediated cell-cell fusion, and pharmacology and resulted in the identification of two novel allosteric modulators of MFSD2A, which are two soluble fragments of SYNC2, namely SYNC2su-co and SYNC2su-co-2, representing first-in-class molecules to inhibit MFSD2A LPCs uptake and increase transcytosis rate.

The invention discloses application of a gp130 inhibitor in preparation of a medicine for treating virus infection, and relates to the technical field of medicines. Experiments prove that combination of IL-6 family cell factors and a gp130 receptor is inhibited, so that downward signal transmission of a JAK2 / STAT3 signal channel triggered by the IL-6 family cell factors is inhibited, the purposes of inhibiting cell fusion induced by SARS-CoV-2 spike protein and further influencing replication and propagation of SARS-CoV-2 are achieved, and a small-molecule inhibitor SC144 is an inhibitor of gp130, has a remarkable effect of inhibiting SARS-CoV-2 virus replication and has a good application prospect. Meanwhile, the traditional Chinese medicine composition also has a remarkable effect of inhibiting replication of viruses such as human immunodeficiency virus, herpes simplex virus and respiratory syncytial virus.

This invention provides a vaccinia virus that induces cell fusion between infected cells and a method for producing the same. Such vaccinia virus is deprived of the K2L gene or the HA gene or functions of the K2L gene and the HA gene and is mutated to induce cell fusion between infected cells and induce cell death.

The application discloses a single-chain antibody, the amino acid sequence of which is shown as SEQ ID NO:1, and the nucleotide sequence of which is shown as SEQ ID NO:2. The single-chain antibody against rabies virus is screened from an antibody library of immunized mice by using a phage display technology. Compared with a traditional cell fusion technology, the phage display technology is simple in operation, low in cost, short in time, and high in antibody discovery and screening efficiency. The single-chain antibody obtained by the application is low in molecular weight and immunogenicity, high in antibody specificity and affinity, and has the activity of neutralizing rabies virus, thereby providing a new idea for basic research of rabies virus and development of therapeutic drugs.

The invention belongs to the technical field of cell cryopreservation, and particularly relates to a cell cryopreservation solution, application thereof and a cell cryopreservation method. A death inhibitor and a solvent are selected for specific cells, and the cell death process related to cytolytic organelle membrane damage is intervened from the molecular level, specifically, the solvent is used for reducing ice crystal formation inside and outside the cells, the membrane protection effect is exerted to achieve the high resuscitation motility rate, a programmed death pathway is further blocked in a targeted mode through the death inhibitor, and the cell death effect is improved. The programmed death after cell resuscitation is relieved, the delayed death rate of the resuscitated cells is reduced, the long-term cell fusion degree of the resuscitated cells is improved, and the long-term survival rate of the resuscitated cells is improved.

The present application relates to a hybridoma cell strain secreting benzyl chlorophenol monoclonal antibody and its application, and belongs to the technical field of immune detection. The present application synthesizes benzyl chlorophenol hapten, prepares benzyl chlorophenol complete antigen, and through the steps of mouse immune response and cell fusion, a hybridoma cell strain secreting benzyl chlorophenol monoclonal antibody is screened and named CBP-3D4, which is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC NO.46222. The monoclonal antibody secreted by the hybridoma cell strain has good sensitivity and specificity, the IC 50 value of the monoclonal antibody secreted by the hybridoma cell strain for benzyl chlorophenol is 97.741 ng / mL, the cross reactivity of the monoclonal antibody for various benzyl chlorophenol functional analogs is less than 1%, and the monoclonal antibody can be applied to the preparation of benzyl chlorophenol detection products to realize the rapid quantification of benzyl chlorophenol.

The invention discloses a stem cell-based microvesicle as well as a preparation method and application thereof, belongs to the technical field of biological tissue engineering, and particularly relates to a preparation method of the stem cell-based microvesicle, which comprises the following steps: culturing stem cells until the cell fusion degree is 80-90%, performing hypoxia treatment, and cleaning for 2-3 times; standing and culturing the cleaned stem cells, then adding a calcium ion carrier, and carrying out ultrasonic treatment; the serum-free basic culture medium containing the calcium ion carrier is immediately absorbed and removed, the cells are cleaned through PBS, culture is conducted for 24 h, and cell supernatant is collected and merged respectively; performing first centrifugation to remove cell debris; performing second centrifugation to remove dead cells and large cell debris; carrying out third centrifugation to remove organelles; fourth centrifugation is carried out, large vesicles are removed, and a final supernatant is obtained; and filtering and centrifuging to obtain a microvesicle precipitate. The preparation method of the microvesicles aims at systematically improving the yield and biological activity of the stem cell microvesicles through low-oxygen pre-stimulation, ultrasonic-calcium ion carrier synergistic induction and differential centrifugation.

The present application provides a method for directional transfer of heterologous chromosomes, which comprises the following steps: artificially adding a functional intron containing a loxP sequence into a drug resistance gene, splitting the drug resistance gene into two parts with an overlapping region containing a loxP sequence, inserting the two parts of the drug resistance gene into target chromosomes of a donor species and a recipient species respectively (the two parts of the drug resistance gene undergo Cre / loxP site-specific recombination under the action of a Cre recombinase, and the drug resistance function is recovered), and realizing directional transfer of heterologous chromosomes by means of heterologous cell fusion. The method can be widely applied to gene function research, construction of human disease models, development of therapeutic antibodies, and pharmacological and pharmacodynamic tests.

The present application belongs to the field of plant biotechnology, and particularly relates to a Korean pine cell fusion method based on chemical induction combined with a magnetic bead method. The method comprises the following steps: S1, preparing first protoplasts and second protoplasts; S2, preparing passivated first protoplasts and passivated second protoplasts; S3, mixing the passivated first protoplasts with the passivated second protoplasts in equal volumes after the passivated first protoplasts are cultured with a sodium silicate aqueous solution to obtain mixed protoplasts; S4, performing electrofusion on the mixed protoplasts in a cell fusion instrument to obtain fusion products; and S5, screening hybrid cells from the fusion products by using a magnetic bead method. The present application adopts a double screening mechanism of chemical passivation and magnetic bead screening, inactivates parent cells by treating with iodoacetamide and rose Bengal B, and then specifically enriches hybrid cells by using the magnetic bead method, so that the acquisition rate and screening accuracy of hybrid cells are significantly improved.

The invention discloses a preparation method of uterine blood mesenchymal stem cells, and relates to the technical field of uterine blood mesenchymal stem cell preparation, and the preparation method comprises the following steps: filtering and diluting a uterine blood sample, adding the diluted uterine blood sample into a sample density separation medium, centrifuging, collecting an upper layer and a middle layer, repeatedly centrifuging, and taking cell precipitate to obtain primary cells of the uterine blood mesenchymal stem cells; resuspending, culturing until the fusion degree of the cells reaches 70-90%, digesting by using a recombinant trypsin solution, collecting a cell suspension, centrifuging, resuspending, inoculating and culturing until the fusion degree reaches 70-90%, digesting by using a recombinant trypsin solution, centrifuging, and resuspending; centrifuging for two times, taking cell precipitate, and resuspending by using a cell freezing medium to obtain the uterine blood mesenchymal stem cells. According to the method, after the uterine blood sample is filtered, the centrifugation step is omitted, and the uterine blood sample is directly added to the upper layer of the sample density separation liquid for centrifugal extraction, so that the normal living cell proportion of primary cells and the adherence rate after inoculation are obviously improved, the production cycle is obviously shortened, and the yield of the primary cells is improved.

The present application relates to a hybridoma cell strain secreting clozapine and its metabolite monoclonal antibody, and belongs to the technical field of immune detection. The present application prepares a hapten by chemically synthesizing clozapine and its metabolite, and then prepares a complete antigen, and then screens a hybridoma cell strain secreting clozapine and its metabolite monoclonal antibody through animal immunization and cell fusion. The hybridoma cell strain LDP-2B10 provided by the present application has been preserved in the China General Microbiological Culture Collection Center (CGMCC) with a preservation number of CGMCC No. 46233. The monoclonal antibody secreted by the hybridoma cell strain has good sensitivity and specificity, the IC 50 value of the monoclonal antibody for clozapine is 5.041 ng / mL, the IC 50 value of the monoclonal antibody for the metabolite N-desmethylclozapine is 5.524 ng / mL, and the IC 50 value of the monoclonal antibody for the metabolite clozapine-N-oxide is 5.551 ng / mL. The cross reactivity of the monoclonal antibody for various clozapine functional analogues is less than 1%, and the monoclonal antibody can be used for detecting clozapine in urine and blood drug concentration, and has practical application value.

The application relates to a single cell recognition method and a storage medium, wherein the single cell recognition method comprises the following steps: obtaining single cell original gene expression data, a gene label control table and pre-training prior gene feature data; obtaining correction data based on the single cell original gene expression data, generating a cell supervision label, combining the gene label control table to construct a single cell gene set activity matrix; generating a training data set based on the above matrix, label, prior data and correction data, inputting the training data set into a classifier and a reconstruction module of an initial cell deep learning network, obtaining a classification result and a constructed activity matrix; obtaining a total loss by weightedly combining a label loss and a reconstruction loss, iteratively training a gradient in a reverse direction, and generating a cell deep learning network; inputting the correction data into the network, inferring a cell fusion embedding result, and further obtaining a single cell recognition result. Through the application, the problem of low single cell recognition accuracy is solved.

The invention belongs to the technical field of nano medicine, and particularly discloses a method for obtaining exosomes with different cell biological functions, which comprises the following steps: preparing hybrid cells HCs integrating different cell functions by a cell fusion method, and further collecting exosomes H / Exos derived from the hybrid cells. The invention also discloses the exosome and application of the exosome in preparation of drugs. Different hybrid cells have multiple functions of cells before fusion, and exosomes secreted by the hybrid cells inherit the functions of the hybrid cells. The hybrid cells can continuously secrete exosomes with the characteristics of the two parent cells in the proliferation process. The defect that the end-to-end differentiated cells cannot be proliferated is overcome, so that large-scale preparation of the natural multifunctional exosome can be realized, and a new method is provided for customized exosome preparation for disease treatment.

The invention relates to a hybridoma cell strain secreting a dimethyl yellow monoclonal antibody and application of the hybridoma cell strain, and belongs to the technical field of immunodetection. 4-(4-aminophenyl) butyric acid and 4-dimethylphenol are used as raw materials to prepare a dimethyl yellow hapten through a diazo method reaction, a dimethyl yellow complete antigen is further obtained, a hybridoma cell strain is obtained through cell fusion and screening of an indirect competitive enzyme-linked immunosorbent assay, the cell strain secretes a dimethyl yellow monoclonal antibody, and the monoclonal antibody can be used for preparing the dimethyl yellow monoclonal antibody. The kit has good detection specificity and detection sensitivity on dimethyl yellow, has an IC50 value of 1ng / mL, can be applied to preparation of an immunodetection product of dimethyl yellow, and provides a brand new efficient tool for detection of dimethyl yellow.

The application discloses a method for preparing stem cell exosomes by using multiple pulse electric field stimulation, which comprises the following steps: step 1, thawing the frozen stem cells and preparing cell suspension, and adjusting the cell density to a first cell density preset value; step 2, when the cell fusion degree in the cell suspension reaches a first cell fusion degree preset value, the cell suspension is subjected to digestion treatment; step 3, placing the cell suspension after digestion treatment in an electric conversion cup, and using a nanosecond pulse electric field with a preset electric field intensity to stimulate the cell suspension in the electric conversion cup; step 4, collecting the cell supernatant in the culture dish for exosome extraction within a first preset time length after the nanosecond pulse electric field pretreatment, meanwhile, adding culture medium in the culture dish again and culturing for a preset time length; and step 5, repeating steps 2 to 4 until the cells reach a preset generation number.

The invention relates to the technical field of antibodies, in particular to a monoclonal antibody generated by a strychnine hybridoma cell strain and application of the monoclonal antibody. The strychnine hybridoma cell strain is preserved in China General Microbiological Culture Collection Center (CGMCC), the address is Institute of Microbiology, Chinese Academy of Sciences, No.3, No.1 Yard, Beichen West Road, Chaoyang District, Beijing, the strychnine hybridoma cell strain is classified and named as the strychnine hybridoma cell strain, the preservation date is June 5, 2025, and the preservation number is CGMCC No.46542. According to the present invention, strychnine is adopted as a raw material to prepare an artificially synthesized antigen having immunocompetence and characteristics, a mouse is induced to produce a strychnine specific antibody through an immunization mode, and then the hybridoma cell strain capable of specifically recognizing strychnine is prepared through cell fusion; the produced monoclonal antibody is successfully applied to a strychnine immunoaffinity column, a strychnine enzyme-linked immunosorbent assay kit, a strychnine colloidal gold immunochromatography detection card and other products.

The invention relates to the technical field of biological medicines, and particularly discloses an anti-PRRX2 monoclonal antibody and application thereof in detection of gemcitabine drug resistance of adenocarcinoma. The antibody is prepared through a hybridoma technology, a BALB / c mouse is immunized with a recombinant human PRRX2 protein, and a high-affinity antibody strain is obtained through cell fusion, screening and monocloning. Sequences of light and heavy chain variable regions of the antibody are shown as SEQ ID No: 7 and SEQ ID No: 8, and sequences of a CDR region are shown as SEQ ID No: 1-6. After the antibody is coupled with HRP, the antibody is used for Western Blot to detect PRRX2 expression in a pancreatic ductal adenocarcinoma (PDAC) patient sample, and the result shows that the detection sensitivity to a gemcitabine drug-resistant sample reaches 96.6%, and the specificity is 100%. The antibody disclosed by the invention has high specificity and stability, provides a reliable tool for clinical drug resistance detection, and is beneficial to formulating an individualized treatment strategy.

The invention discloses a monospecific monoclonal antibody for resisting zearalenone-14 glucoside and application of the monospecific monoclonal antibody. An immunogen and a coating antigen are used; the monoclonal antibody capable of monospecifically recognizing the zearalenone-14 glucoside and the rapid detection kit for detecting the zearalenone-14 glucoside are obtained through the technologies of mouse immunization, serum detection, cell fusion, monoclonal screening, ascites induced antibody and the like. The method is suitable for single-specificity trace residue detection of the zearalenone-14 glucoside in samples such as corn, rice, millet, wheat and oat, has very high recognition sensitivity to the zearalenone-14 glucoside, and realizes rapid, large-batch and low-cost detection of the zearalenone-14 glucoside.

The invention belongs to the technical field of virus detection, and particularly relates to a porcine delta coronavirus N protein monoclonal antibody and a latex immunochromatography test strip containing the antibody. According to the invention, a prokaryotically expressed PDCoV N protein is used as an immunogen to immunize a mouse, and a specific monoclonal antibody 2G3 and a specific monoclonal antibody 9C4 are screened through cell fusion and a subcloning technology. The test strip comprises a PVC (polyvinyl chloride) bottom plate, a sample pad, a combination pad, a nitrocellulose membrane (NC membrane) and a water absorption pad, a quality control line C and a detection line T are respectively arranged on the NC membrane. The test strip can specifically recognize PDCoV, the lowest detection limit can reach 103.0 TCID50 / mL, the test strip has no cross reaction with other clinical common diarrhea viruses, the result interpretation only needs 15 min, and the test strip has the advantages of being rapid, sensitive, easy and convenient to operate and the like.

The present invention discloses an anti-giant panda CDV monoclonal antibody, hybridoma cell line, and uses thereof, belonging to the field of biotechnology. The amino acid sequence of the heavy chain variable region of the monoclonal antibody is shown in SEQ ID NO. 2, and the amino acid sequence of the light chain variable region is shown in SEQ ID NO. 4. Using cell fusion technology, the present invention obtained the hybridoma cell line 4A3 that secretes anti-giant panda CDV monoclonal antibodies. The cell line culture supernatant was further collected and purified to obtain a highly specific monoclonal antibody.

Disclosed herein include methods, compositions, and kits suitable for use in e.g., nucleic acid delivery. Provided are compositions (e.g., nucleic acid compositions) comprising polynucleotide(s) encoding a fusion protein, a dimerization fusion protein, a soluble RBP, an adapter fusion protein, and / or a cell fusion protein. The compositions also comprise one or more polynucleotides comprising one or more RNA cargo molecules. The fusion protein, dimerization fusion protein, and / or adapter fusion protein can comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD). Recruitment of one or more ESCRT proteins results in secretion of enveloped nanoparticles (ENPs) from a cell in which at least one of a fusion protein, a dimerization fusion protein, and / or an adapter fusion protein are expressed. The ENPs can comprise the one or more RNA cargo molecules. There are also provided populations of ENPs in some embodiments.

The invention discloses an anti-human MBP monoclonal antibody and an application thereof. A complementarity determining region of a heavy chain variable region of the monoclonal antibody has an amino acid sequence as shown in SEQ ID NO.1-3, and a complementarity determining region of a light chain variable region of the monoclonal antibody has an amino acid sequence as shown in SEQ ID NO.4-6. The monoclonal cell line capable of efficiently secreting the anti-human MBP monoclonal antibody is obtained by taking chemically synthesized polypeptide as an immunogen to immunize a mouse and carrying out cell fusion, screening and subcloning. The antibody obtained by the invention has high specificity and high binding capacity, can effectively bind MBP protein from serum in a targeted manner, and is suitable for detecting MBP protein in blood, so that nerve injury diseases possibly causing MBP protein concentration change are evaluated, and more convenient conditions are provided for diagnosis of mild traumatic brain injury.

The present application relates to a kind of monoclonal antibody that can identify human smooth muscle myosin antigen, preparation method and its use in immunodetection.The above technical solution selects the amino acid of 1180-1400 of smooth muscle myosin as antigen peptide, codon optimization becomes the gene fragment suitable for expression in escherichia coli, and finally the recombinant protein includes smooth muscle myosin fragment and histidine protein label.The recombinant protein is immunized to mouse, and is subjected to cell fusion, screening and subcloning, obtains mouse hybridoma cell strain 15C1D4 and the anti-smooth muscle myosin monoclonal antibody secreted by the cell strain.The antibody obtained by the present application has high specificity, sensitivity, and can specifically recognize the cell expressing smooth muscle myosin, and is suitable for immunological detection, especially immunohistochemical detection.

The present application relates to the field of cell culture, in particular to a culture and screening method of MDCK monoclonal cell strain, and provides a culture method of MDCK monoclonal cell strain, comprising the following steps: S1, culturing pretreated cells to logarithmic phase; S2, inoculating the cells in logarithmic phase in S1 to the first culture carrier after diluting to a concentration of 1 cell / 100 μL; S3, inoculating the single cell colony in the first culture carrier in S2 to the second culture carrier after digestion, until the cell fusion degree reaches 70-80%, and obtaining the MDCK monoclonal cell strain. The present application realizes early detection without expanding the monoclonal cells to T75 culture flask, avoids the step of gradually expanding and culturing for 7-10 days in the traditional method, greatly shortens the overall screening period from the original 14-21 days, and can quickly match the demand of H3N2 virus variation for highly adaptive cell strains.