60 results about "Syncytial cell" patented technology

The invention discloses a multi-targeting fusion cell membrane modified bionic nano delivery system as well as a preparation method and application thereof. The multi-targeting fusion cell membrane modified bionic nano delivery system comprises a fusion cell membrane and a negative electricity nano core wrapped by the fusion cell membrane, wherein the fusion cell membrane is obtained by fusing a macrophage membrane and a cancer cell membrane; and the negative electricity nano core carries a hydrophobic drug. The surface of the fusion cell membrane contains proteins on the surfaces of a macrophage membrane and a cancer cell membrane, so that the fusion cell membrane has a targeted adhesion recognition function, and a modified nano drug loading system is endowed with a multiple tumor targeting function. The fusion cell membrane modified bionic nano delivery system can deliver chemotherapeutic drugs, has the characteristics of good stability, low immunogenicity and good biocompatibility,and provides an effective means for hydrophobic therapeutic drug delivery in tumor targeted therapy.

The invention discloses a GPC3 (glypican-3) monoclonal antibody hybridoma cell strain 7D11 and a preparation method and application thereof, which belong to the technical field of bioengineering. The GPC3 monoclonal antibody hybridoma cell strain 7D11 has a preservation code of CGMCC No.5426. The preparation method of the hybridoma cell strain 7D11 includes: using GPC3 protein as an immunogen to immunize a mouse, subjecting spleen cells of the mouse with serum titer more than 1:104 and myeloma cells SP2 / 0 to fusion, using a HATRPMI-1640 medium to screen fused cells, screening by ELISA (enzyme-linked immuno sorbent assay) and multiple limiting dilution process, and finally obtaining the hybridoma cell strain 7D11. The GPC3 monoclonal antibody hybridoma cell strain 7D11 is high in yield of secreted antibodies and easy to survive, the secreted monoclonal antibodies are high in titer, flexible in reaction, easy to detect, low in production cost and widely applicable to detection of GPC3 protein expression.

Provided are assays and methods for creating proto-tissues from aggregates of cells. The invention concerns assays and methods useful in tissue engineering and reconstruction techniques, specifically in the formation of macrotissues from microtissues using microtissue pre-culture time as a controlling parameter.

The present invention relates to methods and treatment protocols for the immunotherapy of cancer by administering a therapeutically effective dose of fusion cells formed by fusion of autologous dendritic cells and autologous non-dendritic cells in combination with interleukin-12.

The invention relates to a B cell screening method and application of the B cell screening method in preparing a monoclonal antibody. The B cell screening method comprises the steps of S11, adding a signal peptide sequence of secretory expression at the N end of a Protein A protein gene, and adding a membrane-spanning gene sequence at the C end to obtain a fused exogenous gene; S12, using the fused exogenous gene to conduct transfection on a B cell of which immunity is completed, and adopting the B cell as a cell to be screened; S13, adding an antigen for labeling fluorescent into the cell to be screened, using a flow cytometry to sort and collect cell carrying fluorescent to obtain a positive B cell. According to the B cell screening method and application of the B cell screening method in preparing the monoclonal antibody, antibody molecules are gathered on the surface of the cell, through a fast and precise sorting function of the flow cytometry, a cell to be fused is screened in advance, thus the success rate and accuracy rate of a traditional mode are sharply increased, and the problem of detection of a monoclonal antibody cell strain in a later period is solved at the same time.

The invention discloses a GPC3 (glypican-3) monoclonal antibody hybridoma strain 8G6, and a preparation method and application thereof, which belong to the technical field of cell engineering. The preservation code of the GPC3 monoclonal antibody hybridoma strain 8G6 is CGMCC (china general microbiological culture collection center) No.5427. The preparation method of the GPC3 monoclonal antibody hybridoma strain 8G6 includes: using GPC3 protein as immunogen to immunize a mouse; and fusing mouse splenocytes having serum titer more than 1:104 with SP2 / 0 myeloma cells, using an HATRPMI-1640 medium to screen fusion cells, and finally obtaining the hybridoma strain 8G6 by ELISA (enzyme-linked immunosorbent assay) and repeated limiting dilution. The GPC3 monoclonal antibody hybridoma strain 8G6 is high in yield of secretory antibodies, and is easy to survive, and the secreted monoclonal antibodies are high in titer, sensitive in reaction, easy to detect, low in production cost and widely applicable to detection of GPC3 protein expression.

The present invention provides in one aspect novel fusion partner cells that ectopically express one or more genes that alter the phenotype of a hybrid cell made from a fusion of the fusion partner cell and a fusion cell, hybrid cell lines produced using the fusion partner cells. The invention in another aspect provides antibodies produced by certain hybrid cell lines, and compositions containing one or a combination of such antibodies or antigen-binding fragments thereof. The invention also provides in another aspect methods of using the antibodies or antigen-binding fragments thereof for diagnosis and treatment of diseases characterized by the antigens specifically bound by the antibodies.

It is intended to provide a monoclonal antibody that specifically recognizes human soluble LOX-1, particularly a monoclonal antibody with a dissociation constant (Kd) for human soluble LOX-1 of 1×10−9 (M) or less. The antibody can be produced from a hybridoma prepared by a method including the following steps of: (1) immunizing a non-human animal with a prokaryotic cell-derived human LOX-1 extracellular domain; (2) collecting antibody-producing cells from the animal; (3) fusing the antibody-producing cells with myeloma cells; (4) selecting hybridomas that produce a monoclonal antibody reacting with the human LOX-1 extracellular domain from the fused cells obtained in the above step; and (5) selecting a hybridoma that produces a monoclonal antibody reacting with a eukaryotic cell-derived human LOX-1 extracellular domain from the selected hybridomas.

The invention belongs to the field of pharmaceutical preparations. The invention relates to a uricase / catalase lipid nanoparticle wrapped by a fusion cell membrane and a preparation method of the uricase / catalase lipid nanoparticle. According to the prepared fusion cell membrane coated uricase / catalase lipid nanoparticles, the stability of uricase can be improved, the protease hydrolysis resistance of uricase is improved, the bioavailability is improved, the inflammation chemotactic ability is enhanced, and the toxic and side effects are reduced.

The invention relates to a method for producing a pure human-derived monoclonal antibody by mixed cell culture, which comprises the following steps: co-culturing T-cells, B-cells and human lymph node endothelial cells, and activating specific immunological cells with antigen to promote B-cell division and proliferation, thereby obtaining a monoclonal antibody cell strain with antigenic specificity; and obtaining the pure human-derived monoclonal antibody. The technique adopted by the invention is a mixed cell culture method, is simple and easy to implement and convenient to operate, settles a matter at one go, has a simple screening process, and saves substantial human and material resources; by using the invention, the production efficiency of the monoclonal antibody is greatly enhanced, and the yield of producing antibody from fusion cells is multiplied; and most cloning successfully implements the transformation from Type IgM to IgG, so that the production probability of the IgG type monoclonal antibody is greatly increased.

The present invention relates generally to the field of tissue engineering and more particularly to a method for generating tissue suitable for use in tissue replacement and / or tissue rejuvenation therapy and / or as a source of cell-derived therapeutic or diagnostic agents including proteins and hormones. Even more particularly, the present invention contemplates the use of cell fusion techniques involving single cell, mini-bulk or macro-bulk cell fusion to generate tissue or cells useful for tissue replacement and / or tissue rejuvenation therapy or a range of organs or areas of the body. The resulting tissue or cells may also secrete or generate a range of cytokines, enzymes, hormones and the like which have improved or more efficacious properties relative to analogous molecules produced from non-fused cells. The present invention further provides an apparatus having aspects controlled by data processing means which facilitates the fusion of a pair of cells. Of the pair of cells, at least one of the cells in the pair may be a mature cell or is capable of differentiating or developing into a mature cell. The subject invention further provides isolated molecules such as cytokines, receptors, antibodies, hormones, heat shock proteins, enzymes, and glycoproteins such as mucins, lectins and heparan sulfates derived from fused cells. These molecules may be characterized by having altered glycosylation patterns, altered post-translational modifications, greater activity, being more efficacious or being more stable relative to analogous molecules from non-fused cells. The present invention further provides novel cell fusates or cell hybrids having a pattern of cell surface markers unique relative to the at least two cells which fuse together to generate the cell. These cell markers are useful in selecting particular cell hybrids and as proprietary tags.

The invention discloses a bionic Janus magnetic-mesoporous silica nanoparticle for the specific capture of CTCs, and a preparation method and application thereof. The preparation method comprises thefollowing steps: 1) preparing a carboxylated Janus magnetic-mesoporous silica nanoparticle; 2) preparing a fusion cell membrane; and 3) coating the surface of the Janus magnetic-mesoporous silica nanoparticle with the prepared fusion cell membrane to obtain the bionic Janus magnetic-mesoporous silica nanoparticle. In the bionic Janus magnetic-mesoporous silica nanoparticle, the core, namely the Janus magnetic-mesoporous silica nanoparticle, has an asymmetric structure and a rod-like morphology, and has a large specific surface area; and the surface of the Janus magnetic-mesoporous silica nanoparticle is coated with the fusion cell membrane, the non-specific adsorption of blood cells can be reduced with the help of the characteristic that leukocytes cannot be aggregated in the circulatory system, and thereby the bionic Janus magnetic-mesoporous silica nanoparticle can be well applied to the detection of CTCs.

The invention discloses a high sensitive fusion cell screening method, belongs to the field of gene engineering, and relates to a fusion cell quick screening scheme based on Cre recombinase and Loxp sequence specific recognition. Through incision of the Cre recombinase and the Loxp sequence, fluorescence switching and cell resistance switching of the fusion cell are achieved, and therefore singlecell screening and subsequent expanding culture of the fusion cell are achieved. Universal expression Cre protein nude mice are used for injecting modified tumor cells into the mouse bodies so that tumor cell metastasis and development caused by cell fusion function can be observed. The invention provides a lentivirus plasmid vector, wherein the nucleotide sequence of a fluorescence imaging analysis carrier A which is capable of achieving fusion cell high sensitive rapid screening and in vivo tumor cell metastasis is shown as Seq ID No.1, and the the nucleotide sequence of a carrier B is shownas Seq ID No.2.

The invention discloses a fusion cell membrane nano-vesicle for tumor immunotherapy as well as a preparation method and application of the fusion cell membrane nano-vesicle. The fusion cell membrane nano-vesicle is formed by fusing two or more cell membrane vesicles from cells of different sources, and the cell membrane vesicles from the cells of different sources overexpress receptors corresponding to different immune checkpoints. The fusion cell membrane vesicle does not need complex design and preparation, and does not bring immunotoxicity; compared with the cell membrane vesicles with a single target marker, the fusion cell membrane nano-vesicle has the advantages that the treatment effect is improved, and compared with cocktail therapy in which the cell membrane vesicles with different target markers are simply mixed and used, the fusion cell membrane nano-vesicle has the advantages that the treatment effect is also improved; besides, a single cell membrane vesicle component in the fusion cell membrane vesicle can be customized independently, so that the structure has high degree of freedom, and various different tumor cells can be selectively targeted; and due to homologous targeting and immune antigens of tumor cell membranes, the treatment effect on various tumors can be improved at the same time.

A hybridoma cell strain secreting a nifursolol residue marker monoclonal antibody with CGMCC No. 14698 is prepared in the following way: BALB / c mice are subjected to the first immunization with a complete Freund's adjuvant, subjected to booster immunization with an incomplete Freund's adjuvant for four times, and subjected to rush immunization once with nifursolol residue marker complete antigen without a Freund's adjuvant so that the BALB / c mice are immunized; the spleen cells of the immunized mice with high titer and low IC50 were fused with mouse myeloma cells by a PEG method, and the fused cells are screened through indirect competitive ELISA and subcloned three times. The monoclonal antibody secreted by this cell strain has good specificity and detection sensitivity (IC50 value of 2 μg / L) to the nifursolol residue marker and can be used for residue detection of the nifursolol residual marker in food.

This invention provides methods for determining the ability of a gene construct to alter the rhythm and contractility of a syncytial cell. Furthermore, this invention provides methods for constructing a gene construct capable of altering the rhythm or contractility of a syncytial cell. Finally, this invention provides a method for constructing a gene construct capable of coupling to a syncytial cell.

The invention belongs to the technical field of medicinal preparations, and relates to a mesenchymal stem cell-based fusion cancer cell membrane bionic nanoparticle, in particular to a mesenchymal stem cell-based tumor and bone metastatic tumor targeted fusion cancer cell membrane bionic nanoparticle (TB@NP) and a preparation method and application thereof. The bionic nanoparticle is a drug-loadednanoparticle, and is coated with a layer of fusion cell membrane; the fusion membrane consists of a mesenchymal stem cell membrane and a cancer cell membrane, and is made into a bionic nano drug delivery system capable of targeting tumors and tumor bone metastatic parts. The bionic nanoparticle can be an inorganic nanoparticle and organic nanoparticle, and entrapped drugs can be gene drugs, smallmolecule drugs and the like. In-vivo and in-vitro experiments prove that the bionic nano delivery system can accurately deliver drugs to tumors and tumor bone metastatic parts, is high in targeting property and high in safety, can enhance the curative effect of the drugs and reduce toxic and side effects, and has relatively high clinical practice significance.

The present invention provides a lentiviral vector system having a higher titer, while sustaining an excellent retrograde transport ability, particularly, in the brain.The present invention also provides a kit for preparing a retrograde transport viral vector comprising:(1) a packaging plasmid containing the gag gene and the pol gene of HIV-1;(2) a packaging plasmid containing an accessory gene of HIV-1;(3) a transfer plasmid containing an target gene (a transgene); and(4) an envelope plasmid containing, as an envelope gene, a gene encoding a fused polypeptide comprising a fused extracellular domain consisting of the N-terminal region of an extracellular domain of rabies virus glycoprotein (RV-G) and the C-terminal region of an extracellular domain of vesicular stomatitis virus glycoprotein (VSV-G), a transmembrane domain of RV-G or VSV-G, and an intracellular domain of VSV-G, and the like.

A novel sugar alcohol-producing yeast strain genome rearrangement technology mainly includes high throughput screening of a sugar alcohol-producing yeast strain on the basis of a color development method and fusion cell high-efficient flow sorting on the basis of bi-fluorescent staining. By means of the technology, high-effective genome rearrangement and screening of alcohol-producing yeast are achieved. The technology is simple, low-cost, high-efficient and practical. Through the technology, high-efficient and quick improvement on natural pichia pastoris being abnormal in sugar alcohol producing can be successfully achieved, thereby increasing the yield of sugar alcohol by more than 34%.

The invention discloses a preparation method of a mycoplasma synoviae monoclonal antibody. The conservative analysis is carried out on two imaginary membrane proteins with the mycoplasma membrane surface protein sizes of 20Kda and 13Kda; then recombinant plasmids synthesizing, protein inducible expression, and affinity chromatography purification are carried out; the purified recombinant protein is used as immunogen to immunize BALB / c mice, cell fusion is carried out after antibody detection is qualified, a single monoclonal antibody cell strain is obtained after subcloning of fusion cells, and the prepared monoclonal antibody can well and specifically recognize mycoplasma synoviae antigen.

The invention provides a human apolipoprotein AV monoclonal antibody and an application thereof. The human apolipoprotein AV monoclonal antibody is obtained by syncretizing a splenic cell and a myeloma cell of a human apolipoprotein AV immunized mouse and then cultivating the syncretized cell. The monoclonal antibody can be used for detecting human apolipoprotein AV in human serum. The Apo AV monoclonal antibody can be used for detecting Apo AV in human serum. When the Apo AV monoclonal antibody is used for detecting the Apo AV, the linear detection scope is wide, ApoAI, ApoB and Lp(a) in the serum do not influence a detection result, and the detection result has good stability, therefore, the Apo AV monoclonal antibody can be used for preparing corresponding detection products so as to detect the Apo AV concentration in the human serum, determine the high risk group of coronary heart disease as early as possible and prevent coronary heart disease in time and cure the coronary heart disease in the early period.

The present invention is concerned with a system for fusion at least two cells. The system has an optical tweezers system for generating moving optical traps for manipulating the cells, an optical scissors system for cutting cell membrane of the cells and inducing fusion of the cells, an incubation system for providing cell culture in which the cells suspend and cell culture environment for the cells, and a visual detection system allowing visual monitoring of the cells undergoing fusion.

The present invention relates to a preparation method of a regenerated gene protein REG1 alpha monoclonal antibody. The method comprises the following steps: (1) preparing immunized mice; (2) culturing myeloma cells for 3-4 days to obtain cultured mouse myeloma cells; (3) dissolving REG1 alpha protein, inoculating abdominal cavity of the immunized mice with the dissolved REG1 alpha protein for immunization, obtaining splenocytes, and dispersing the splenocytes to obtain a splenocyte precipitate; (4) centrifuging the cultured mouse myeloma cells to obtain a myeloma cell precipitate; (5) respectively suspending the splenocyte precipitate and the myeloma cell precipitate by using a DMEM high-glucose culture solution, conducting mixing, and centrifuging the mixture to obtain a precipitate; (6)centrifuging and suspending the precipitate to obtain fusion cells; (7) culturing and determining the fusion cells, and determining hybridoma cells; (8) screening and separating monoclonal antibodies; and (9) preparing a large amount of monoclonal antibodies, namely producing the monoclonal antibodies from a hybridoma cell line according to a conventional method. The preparation method is rapid and high in specificity.

The invention discloses a cancer cell / DC (Dendritic Cell) fusion tumor vaccine for expressing Endoglin and a preparation method for the cancer cell / DC fusion tumor vaccine. According to the cancer cell / dendritic cell fusion tumor vaccine for expressing Endoglin and a preparation method for the cancer cell / dendritic cell fusion tumor vaccine, the method for preparing fusion cells comprises the step of fusing tumor cells and dendritic cells to obtain the fusion cells; the tumor cells are recombinant cells for expressing the Endoglin or the recombinant cells which contain the gene for encoding the Endoglin; the Endoglin is the protein of A1) or A2) below: the A1) is the protein of which the amino acid sequence is SEQ ID No.1; the A2) is the protein which has the same function as that of the protein of the A1) and is derived from the protein of the A1) performing substituting and / or deleting and / or adding on one or more amino acid residues of the amino acid sequence which is shown as the SEQ ID No.1.

The invention discloses a heterologous fusion gene modified cancer cell / dendritic cell fusion tumor vaccine and its preparation method. According to the heterologous fusion gene modified cancer cell / dendritic cell fusion tumor vaccine and its preparation method, a method for cell fusion comprises a step of fusing cancer cell which contains Endoglin and alpha1,3 galactosyl transferase related gene and is used for expression of Endoglin and alpha1,3 galactosyl transferase and dendritic cell to obtain fusion cell. The alpha1,3 galactosyl transferase gene encodes protein with its amino acid sequence being SEQ ID No.3. The Endoglin is protein with its amino acid sequence being SEQ ID No.1. The alpha1,3 galactosyl transferase related gene is formed by connecting the alpha1,3 galactosyl transferase gene and GTTI. Nucleotide sequence of GTTI is from the 1st nucleotide to the 548th nucleotide in the SEQ ID No.5 of the sequence table.

The invention relates to a B cell screening method and application of the B cell screening method in preparing a monoclonal antibody. The B cell screening method comprises the steps of S11, adding a signal peptide sequence of secretory expression at the N end of a Protein A protein gene, and adding a membrane-spanning gene sequence at the C end to obtain a fused exogenous gene; S12, using the fused exogenous gene to conduct transfection on a B cell of which immunity is completed, and adopting the B cell as a cell to be screened; S13, adding an antigen for labeling fluorescent into the cell to be screened, using a flow cytometry to sort and collect cell carrying fluorescent to obtain a positive B cell. According to the B cell screening method and application of the B cell screening method in preparing the monoclonal antibody, antibody molecules are gathered on the surface of the cell, through a fast and precise sorting function of the flow cytometry, a cell to be fused is screened in advance, thus the success rate and accuracy rate of a traditional mode are sharply increased, and the problem of detection of a monoclonal antibody cell strain in a later period is solved at the same time.