56 results about "Cell-cell fusion" patented technology

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 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 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 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 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 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 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.

The application provides a cell array capture and pairing microfluidic chip, which comprises cell pair units arranged in an array, each cell pair unit being provided with a large micro-trap, three small micro-traps arranged in a sinking interval in the large micro-trap, three groups of electrode pairs and shielding electrodes between adjacent small micro-traps, and the small micro-traps being located between corresponding electrode pairs. The cell capture and pairing microfluidic chip can capture three kinds of cells on the chip in sequence, realize large-scale triple cell array, and arrange the three kinds of cells in the array in one of each kind of cell and three cells as a group for pairing, so that great convenience and possibility are provided for studying three-cell paracrine and other intercellular interactions or cell fusion.

The application belongs to the field of cell culture. The application provides a culture medium of rat amniotic membrane mesenchymal stem cells and a culture method of rat amniotic membrane mesenchymal stem cells. The culture medium is a complete culture medium, and the culture method is as follows: the rat amnion is cut into tissue blocks smaller than 2mm 2 , is uniformly laid in a culture dish with a spacing of 0.3-0.8cm, complete culture medium is slowly added after the tissue blocks are adhered, the culture medium is supplemented after 12h-36h, the primary medium replacement is performed after 48h-72h of culture, the impurity cells are removed, and the cells are subcultured when the cell fusion degree reaches 80%-90%. The light microscope observation and identification result show that the rat amniotic membrane mesenchymal stem cells cultured by the application basically conform to the cell morphology of rat amniotic membrane mesenchymal stem cells, the CD90&DAPI immunofluorescence identification result shows that the cell purity is greater than or equal to 90%, the cells have the characteristics of stem cells, and the culture method is efficient in amplification.

The invention relates to a diafenthiuron hapten, a monoclonal antibody, a hybridoma cell strain and application thereof, and belongs to the technical field of immunodetection. A diafenthiuron hapten is synthesized, then the diafenthiuron hapten is coupled with protein to obtain a diafenthiuron complete antigen, a hybridoma cell strain secreting the diafenthiuron monoclonal antibody is obtained through the steps of mouse immunization, cell fusion and the like, the hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC NO.46519. The monoclonal antibody provided by the invention has high sensitivity and high specificity to diafenthiuron, the IC50 value is 1.18 ng / mL, and the cross reaction rate to various diafenthiuron analogues is 1t; the method can be applied to preparation of diafenthiuron detection products, and an efficient tool is provided for diafenthiuron immunodetection.

The application belongs to the field of biological medicine and virus infection, and particularly relates to an inhibitor for limiting virus-mediated cell-cell fusion. The application specifically discloses an inhibitor for limiting cell-cell fusion mediated by pseudorabies virus (PRV), wherein the inhibitor contains MARCH1 protein, and the MARCH1 protein includes a derivative thereof, an isolated nucleic acid molecule, a vector or a host cell containing the MARCH1 protein, and / or a fusion. It is found in the research that MARCH1 significantly inhibits the replication of PRV in the stage of cell-cell fusion, that is, it is found in the application that MARCH1 captures a virus protein complex inducing cell-cell fusion in a trans-Golgi network (TGN), and it is proved that MARCH1 has anti-PRV infection activity.

This invention relates to the field of microbial culture, and more specifically to a cell culture system and method. Different cell mixtures are stored separately in different culture dishes. Through drive-controlled adjustment, the different culture dishes are brought together. The brought-to-be-jointed cell mixtures fuse through compression. Peristaltic stirring effectively promotes mixing while avoiding cell damage. After resetting, the cell fusion is observed, and the system continues to combine and fuse different cell types through drive control. This promotes cell fusion, reaction, and storage, facilitating the observation of experimental data and preventing contamination. Multiple culture dishes are evenly fixed on a height-adjusting actuator. Height-adjusting actuators are installed in the central support platform and two side sliding platforms. The beneficial effect of this invention is that it ensures automatic fusion and observation of cells without contamination.

This article describes single-cytokine and multi-cytokine fusion proteins, as well as the VHH cytokine fusion protein. Methods for producing these fusion proteins and their uses are also described. Methods for administering the fusion protein as a single composition to subjects in need are also included.

This invention provides a formulation capable of promoting normal decidualization of the endometrium. An endometrial decidualization promoter or improver containing an effective amount of pyrroloquinoline quinone or its salt is prepared. Furthermore, a cell fusion promoter or improver containing an effective amount of pyrroloquinoline quinone or its salt is also prepared.

The invention belongs to the technical field of biology, and particularly relates to a monoclonal antibody targeting African swine fever virus p30 protein and an epitope of the monoclonal antibody. A mouse is immunized by an African swine fever virus p30 recombinant protein, a cell strain is obtained by utilizing cell fusion and subcloning technologies, and a monoclonal antibody targeting the African swine fever virus p30 protein is generated, a heavy chain variable region of the monoclonal antibody comprises CDR1-3 shown as SEQ ID NO.1-3, and a light chain variable region of the monoclonal antibody comprises CDR1-3 shown as SEQ ID NO.4-6; the monoclonal antibody can specifically recognize and bind to the African swine fever virus, the amino acid sequence of the p30 protein antigen epitope recognized by the monoclonal antibody is shown as SEQ ID NO.11, and the epitope is highly conserved between African swine fever virus gene I type, African swine fever virus gene II type and African swine fever virus I / II type recombinant virus strains, and has important application value in the field of African swine fever virus research and detection analysis.

This invention relates to the field of immunology, and more particularly to a hybridoma cell line and its secreted monoclonal antibody against the metamizole residue marker 4-methylaminoantipyrine (MAA), and its applications. This invention uses 4-aminoantipyrine to react with methyl 4-bromomethylbenzoate, followed by alkaline hydrolysis to obtain a hapten (Formula I). ​​An immunogen is then synthesized and used to immunize mice. After cell fusion and screening, a hybridoma cell line capable of secreting a monoclonal antibody recognizing MAA is prepared, named Metamizole-6A4. This hybridoma cell line has a high efficiency in secreting monoclonal antibodies. The secreted monoclonal antibody can accurately detect MAA, IC50, and other markers. 50 The concentration is 4.06 ng / mL, with high specificity. Cross-reactivity with aminopyrine and its metabolites 4-aminoantipyrine, 4-formamidoantipyrine, 4-acetamidoantipyrine, sulfadiazine, norfloxacin, lincomycin, tylosin, penicillin G, streptomycin, and gentamicin is less than 5%. It can accurately detect MAA residues in food. Formula I.

The invention provides a single-cell multi-omics sequencing data integration method and system based on comparative learning, and the method comprises the steps: single omics feature extraction: employing an intra-omics comparative learning method to capture a local and global potential interaction relationship in single omics data; performing multi-omics feature fusion, realizing inter-omics information alignment by using a cross-modal attention algorithm, retaining feature heterogeneity of each omics, creating positive and negative sample pairs based on a multi-omics pairing relationship, and designing inter-omics comparison loss and inter-omics matching loss to mine an inter-omics correlation relationship; downstream tasks are achieved, and cell type clustering, cell subtype analysis, cell type re-labeling and cell development trajectory prediction are achieved based on cell fusion representation. The influence of high noise, high sparsity and high dimension difference of single-cell multi-omics sequencing data on multi-omics data fusion is relieved, the quality of cell fusion characterization is effectively improved, and help is provided for cell type clustering, cell subtype analysis and cell development trajectory reasoning.

The present application relates to the technical field of biological medicine, and particularly relates to an anti-Aβ 42 Monoclonal antibody, preparation method and application. The present application adopts Aβ 42 protein of human origin as an immunogen to immunize BALB / c mice, extracts B lymphocytes of the spleen of the mice after successful immunization, fuses the B lymphocytes with mouse myeloma cells SP2 / 0 through cell fusion technology, and obtains hybridoma cell strains stably secreting anti-Aβ 42 monoclonal antibody after three rounds of subclone screening, so as to obtain anti-Aβ 42 monoclonal antibody; the anti-Aβ 42 monoclonal antibody can specifically combine with Aβ 42 protein, and can be used for immunological detection, has a broad market prospect, and can have important clinical significance for further development of biological monitoring technology and therapeutic drugs taking Aβ 42 as a target.

The invention relates to the technical field of biological medicine, in particular to an autophagosome for promoting hair growth and an extraction method and application thereof. Comprising the following steps: inoculating human umbilical vein endothelial cells or human placenta mesenchymal cells into a DMEM (Dulbecco Modified Eagle Medium) complete medium, and culturing until the cell fusion degree reaches 80-90%; replacing the DMEM complete culture medium with a serum-free DMEM culture medium, collecting supernate after culture is completed, and centrifuging the supernate to obtain a crude extract containing the autophagosome; and resuspending the crude extract by using PBS, adding an LC3 antibody, and finely extracting by using a paramagnetic particle method to obtain the autophagosome. The method has the advantages of simple operation process, no need of other inducers in the process of inducing the formation of the autophagosome, no too high requirements on laboratories, and high yield of the autophagosome, and provides an efficient, convenient, economical and practical treatment means for alopecia patients.