14 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 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 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 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 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 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 application belongs to the technical field of biological detection, and discloses a capripoxvirus specific competitive ELISA detection kit and application thereof. The application discloses a hybridoma cell strain GTPV-N1P-4H6, and realizes high-efficiency soluble expression of GTPV-N1 based on a prokaryotic expression system. After immunizing mice with GTPV-N1 as an immunogen, a GTPV positive hybridoma cell strain GTPV-N1P-4H6 capable of stably secreting antibodies is successfully prepared and screened after cell fusion. The ELISA titer of the antibody obtained after the monoclonal antibody secreted by the cell strain is purified is 1:64000, the heavy chain of the antibody is IgGa2 subtype, and the light chain is Kappa chain. Based on the antibody, high-efficiency detection of capripoxvirus N1 protein can be realized.

This invention belongs to the field of biodetection technology and discloses a canine distemper virus-specific competitive ELISA detection kit and its application. This invention discloses a hybridoma cell line CDV-H-17E5. Based on a prokaryotic expression system, this invention achieves efficient and soluble expression of CDV H protein. After immunizing mice with this cell line as an immunogen, cell fusion was performed, and a CDV-positive hybridoma cell line CDV-H-17E5 capable of stably secreting antibodies was successfully prepared and screened. The ELISA titer of the antibody obtained after purification of the ascites fluid secreted by this cell line is 1:320,000. The antibody heavy chain is IgGa2 subtype, and the light chain is Kappa chain. Based on this antibody, efficient detection of canine distemper virus CDV H recombinant protein can be achieved.

The application discloses a pig SLPI monoclonal antibody and application thereof, and belongs to the field of biotechnology and cell engineering. The application immunizes mice with a His-SLPI recombinant protein, and carries out cell fusion when the serum titer reaches 1:10000 or above. On the seventh day of the fusion, indirect ELISA is carried out on cell supernatant, and positive wells with higher OD values are screened out. The purified recombinant GST-SLPI protein is used as a coating antigen, the wells are expanded and cultured, and the cells are made into single cells in single wells by using an effective dilution method. After continuous screening to obtain 100% positive wells, the supernatant of the positive cells is used for specific identification of the pig-derived SLPI protein by Western blot and IFA. One hybridoma strain B with a higher binding titer is obtained by identification, the heavy chain amino acid sequence of the SLPI monoclonal antibody produced by the hybridoma strain B is shown as SEQ ID NO. 1, and the light chain amino acid sequence is shown as SEQ ID NO. 2.

This invention discloses an anti-human MBP monoclonal antibody and its applications. The complementarity-determining region (CDR) of the heavy chain variable region of the monoclonal antibody has the amino acid sequences shown in SEQ ID NO. 1-3, and the CDR of the light chain variable region has the amino acid sequences shown in SEQ ID NO. 4-6. This invention uses a chemically synthesized polypeptide as an immunogen to immunize mice. Through cell fusion, screening, and subcloning, a monoclonal cell line that efficiently secretes anti-human MBP monoclonal antibodies is obtained. The antibody obtained by this invention has high specificity and high binding capacity, effectively targeting and binding MBP protein from serum. It is suitable for detecting MBP protein in blood, thereby evaluating neurological injury diseases that may cause changes in MBP protein concentration, and providing more convenient conditions for the diagnosis of mild traumatic brain injury.

This invention belongs to the fields of immunology and in vitro diagnostics, specifically relating to a monoclonal antibody against the S protein of porcine epidemic diarrhea virus (PEDV), its preparation method, and its applications. This invention utilizes a prokaryotic expression system to express and purify the full-length PEDV S protein, using it as an immunogen to immunize mice. Through cell fusion and subcellular screening, a monoclonal antibody targeting the S protein was successfully obtained. The monoclonal antibody provided by this invention can be obtained using conventional genetic engineering or protein engineering methods, avoiding antibody loss during long-term cryopreservation of hybridoma cells. It also facilitates antibody optimization at the gene and protein levels, thereby improving antibody specificity and affinity. The monoclonal antibody prepared by this invention exhibits good conservation, providing materials for the study of PEDV pathogenic mechanisms and the development of PEDV diagnostic kits.

The application belongs to the technical field of biological detection, and discloses a bovine nodular dermatopathy virus specific competitive ELISA detection kit and application thereof. The application discloses a hybridoma cell strain LSDV-ORF132-11H1, and the application realizes high-efficiency soluble expression of LSDV-ORF132 based on a prokaryotic expression system. After mice are immunized by taking LSDV-ORF132 as an immunogen, a LSDV positive hybridoma cell strain LSDV-ORF132-11H1 capable of stably secreting antibodies is successfully prepared and screened after cell fusion. The ELISA titer of the antibody obtained after the monoclonal antibody secreted by the cell strain is purified is 1:64000, the heavy chain of the antibody is an IgGa2 subtype, and the light chain is a Kappa chain. Based on the antibody, high-efficiency detection of bovine nodular dermatopathy virus ORF132 protein can be realized.

The application discloses a method for establishing a goat bone marrow mesenchymal stem cell line and belongs to the technical field of stem cell culture. The application adopts DMEM / F12 to rinse goat bone marrow to obtain a rinsing product; the rinsing product and complete culture medium are mixed and blown to be a single cell suspension, and then inoculation culture is carried out; after the inoculation culture is completed, subculture is carried out; the subculture method is as follows: when the cell fusion degree reaches 80%, the culture medium is discarded, rinsing, digestion and digestion termination are carried out to obtain a digested cell suspension; the digested cell suspension is blown, centrifuged, precipitated and resuspended, and then inoculated into a new culture bottle; after one hour, the non-adherent cell suspension is transferred to a new bottle, and the adherent BMSCs are reserved for continuous culture, so that the subculture is completed; the main cell bank is established by subculture to the third generation; the aging phenomenon that the proliferation speed of BMSCs is slowed down and the differentiation ability is decreased is effectively delayed, and the differentiation potential stability of the BMSCs is ensured.