32 results about "Cell fusion" patented technology

The invention discloses a positive charge fluorescent nanoprobe targeting BCR-ABL fusion protein and application of the positive charge fluorescent nanoprobe in leukemia single cell drug resistance detection, and relates to the field of biological medicine. According to the invention, the surface of the nanoprobe is subjected to specific modification of a polyethylene glycol hydrophilic polymer chain-bridged targeting molecule, so that the functionalized fluorescent nanoprobe with leukemia subcellular oncogenic fusion protein targeting property is successfully constructed. The probe realizes efficient and accurate targeting of the BCR-ABL fusion protein by regulating a subcellular transport pathway, completes diagnosis and quantitative analysis of drug resistance of the leukemia single-cell BCR-ABL fusion protein by utilizing an endocytosis-transport-exocytosis process of cells, can more comprehensively reveal heterogeneity and drug resistance conditions of BCR-ABL positive cells, and has a good application prospect. And a new technical means is provided for accurate diagnosis and treatment of chronic myelogenous leukemia.

This invention discloses an antibody that specifically recognizes Toxoplasma gondii surface antigens and its applications. The invention utilizes hybridoma technology to establish a hybridoma cell line by fusing Balb / C mouse spleen cells with SP2 / 0 cells. Monoclonal antibodies were prepared from these cells using in vitro culture and in vivo ascites induction methods, yielding the monoclonal antibody SH12-S7. This invention provides clinical and research institutions with an antibody that specifically recognizes Toxoplasma gondii surface antigens. It can be used for ELISA assays to detect clinical samples, for immunohistochemical assays to diagnose pathological specimens, and provides a convenient research tool for toxoplasmosis research departments. It has broad application value in the diagnosis and detection of definitive host infections and provides technical support for epidemiological investigations and evaluation of control effects of Toxoplasma gondii.

The application discloses a recombinant protein production and purification method based on ion response controllable phase separation, constructs an engineering cell expressing a fusion protein, the fusion protein comprising a phase separation polypeptide tag and a target protein; the fusion protein is induced to be expressed; the engineering cell is collected, and the cell is crushed in a crushing liquid containing anions, so that the fusion protein is subjected to phase separation and enters an enrichment phase; a supernatant containing host impurities is removed, and a precipitate containing the fusion protein is collected; the concentration of the anions in the precipitate is reduced, so that the fusion protein is redissolved and returns to the supernatant; the supernatant is collected, and a crude extract of the target protein is obtained; wherein the phase separation polypeptide tag comprises at least one continuous five-amino-acid core sequence: X1-X2-X3-X4-X5. The corresponding phase separation peptide, fusion protein and expression vector are also disclosed. The method can reduce the traditional chromatography step, reduce the purification cost, improve the recovery rate and purity of the target protein, and is suitable for the large-scale production of recombinant proteins.

The present embodiments relate to cell confluency estimation and manufacturing. Subject matter of the present embodiments provides computer-implemented methods, computer systems and computer-readable storage media for predicting the results of image-based cell confluency estimation and manufacturing methods.

The application provides an abnormal prothrombin detection kit and application thereof, and relates to the technical field of in-vitro detection.The application provides a kit for abnormal prothrombin detection, which comprises an abnormal prothrombin antibody, wherein the abnormal prothrombin antibody is derived from a hybridoma cell strain 7E8-4-4-1 of fusion of myeloma cells and mouse B cells, and the hybridoma cell strain 7E8-4-4-1 is preserved in the China General Microbiological Culture Collection Center on October 28, 2025, with a preservation number of CGMCC No.46709.The kit has a large detection range, a small lower limit of detection and high accuracy, and has great application potential in the detection of abnormal prothrombin.

The application belongs to the field of stem cell biology and relates to a culture method of a 3D nerve immune organoid containing microglia cells, which comprises the following steps: S1, inducing induced pluripotent stem cells in ectoderm direction and mesoderm direction respectively to obtain nerve-like embryoid bodies and yolk sac-like embryoid bodies; S2, culturing the nerve-like embryoid bodies to make them differentiate into nerves to obtain nerve ring structures, re-digesting the nerve ring structures to obtain nerve progenitor cell single cells, and culturing the yolk sac-like embryoid bodies to make them generate macrophage progenitor cells; and S3, fusing the nerve progenitor cell single cells and the macrophage progenitor cells and continuing to culture to form the 3D nerve immune organoid containing microglia cells. The brain-like organoid of the application can realize the following functions: exploring the control of microglia cells on the proportion of progenitor cells in the development stage, the change of phagocytosis, the control on the number of mature neurons, the influence on cell proliferation and apoptosis, and simultaneously accepting external stimulation and making corresponding functional stress changes.

The invention discloses application of keratin in preparation of a bone repair material, and relates to the technical field of biomedical engineering. The preparation method comprises the following steps: extracting bone marrow mesenchymal stem cells, and carrying out osteogenic induction culture under the intervention of keratin with the concentration of 100-800 [mu] g / ml; the culture conditions of the BMSCs are as follows: in a constant-temperature incubator with the temperature of 37 DEG C and 5% CO2, a culture medium contains a precooled complete culture medium; the first liquid changing time of the primary culture of the BMSCs is 48-72 hours, then the liquid is changed once every 2-3 days, and passage is carried out when the cell fusion degree reaches 80%-90%. Keratin is adopted as a natural biological material, has good biocompatibility and biodegradability and can well interact with cells, and immunological rejection is reduced.

Various examples of the disclosure relate to techniques to count cells in a microscopy image and / or to determine a degree of confluence of the cells in the microscopy image. To that end, machine-learned algorithms are used.

The present invention provides a bone graft composite material and a manufacturing method therefor. The bone graft composite material is applied to wound repair of a user in orthopedic surgery and dental surgery, and is used to promote wound healing of the user, induce growth of an autologous bone cell of the user, and induce fusion of a transplanted bone of the user with the autologous bone cell. The bone graft composite material comprises a high molecular polymer, bone powder, and exosomes. Compared with the prior art, the bone graft composite material provided by the present invention has better osteoinductivity and compatibility, and can stably release biological factors, thereby promoting the repair and regeneration of bone.

This invention discloses a xenogeneic-free culture method for hiPSCs and its quality monitoring method. The xenogeneic-free culture method includes the following steps: hiPSC cells are seeded into the wells of a culture plate coated with VTN-N matrix gel, and cultured for the first stage using E8 complete medium containing 5-15 µM Y-27632. After 20-30 h of culture, the medium is replaced with conventional E8 complete medium without Y-27632 for the second stage of culture, until the cell confluence reaches 60%-80%. This xenogeneic-free culture method ensures efficient cell expansion under conditions with clearly defined chemical composition. This method further optimizes the seeding density and culture period of hiPSCs, controls cell passage at 60%-80% confluence, and significantly reduces the risk of spontaneous differentiation by limiting the maximum culture time and upper limit of confluence.

The invention relates to an MICA and / or MICBmRNA tumor vaccine and application thereof, and provides an MICA and / or MICBmRNA vaccine, a corresponding mRNA construct, a vector, a host cell, a fusion protein, an LNP preparation, pharmaceutical application and a tumor or cancer treatment / prevention method.

A blocking monoclonal antibody screening method based on a porcine PD-1 / PD-L1 monoclonal antibody and a heterologous Fc fusion protein comprises the following steps: (a) preparing a porcine PD-1 or PD-L1 extracellular region recombinant protein; (b) immunizing mice by using the recombinant protein to obtain immune spleen cells; (c) fusing the immune spleen cells with myeloma cells to obtain hybridoma cells; (d) screening and subcloning a hybridoma cell strain capable of secreting an anti-porcine PD-1 or PD-L1 antibody; (e) preparing a porcine PD-L1-rabbit Fc fusion protein; and (f) evaluating the blocking effect of the monoclonal antibody on PD-1 / PD-L1 binding by using the PD-L1-rabbit Fc fusion protein through flow cytometry.

The present invention relates to a novel unconventional method of cell killing for selectively killing or / and inhibiting the growth of target cells. This unconventional method is to kill target cells by persistently over-accumulating organelle-localization sequence fusion antibodies (OLS fusion antibodies) in a key organelle of the target cells with the purpose of clogging or over-occupying the key organelle. The OLS fusion antibodies comprise a cell-binding domain and an organelle-localization sequence (OLS) without any cytotoxic payload. The cell-binding domain of the OLS fusion antibodies functions to selectively bind to a cell-surface antigen of the target cells. The OLS fragment of the OLS fusion antibodies functions to direct the OLS fusion antibodies to a specific organelle of the target cells. The present invention first demonstrates that selectively and persistently accumulating OLS fusion antibodies in a key organelle of the target cells can kill or / and inhibit the growth of the target cells.

The invention provides a benzophenanthrene polycyclic aromatic hydrocarbon hapten, a monoclonal antibody, a hybridoma cell strain and application, and relates to the technical field of immunodetection. According to the invention, triphenylene hapten is synthesized, triphenylene complete antigen is prepared, cell fusion and indirect competitive enzyme-linked immunosorbent assay screening are carried out, a hybridoma cell strain is obtained, the hybridoma cell strain is preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC NO.46732, and the hybridoma cell strain can be used for preparing a triphenylene monoclonal antibody. The monoclonal antibody secreted by the hybridoma cell strain has good detection sensitivity to benzo [a] pyrene, benzo [a] anthracene, benzo [b] anthracene, dibenzo [a, h] anthracene and benzo [k] anthracene, the IC50 values are 6.3 ng / mL, 19.4 ng / mL, 6.8 ng / mL, 55.5 ng / mL and 21.7 ng / mL respectively, and the monoclonal antibody can be used for establishing an immunological detection method for benzophenanthrene polycyclic aromatic hydrocarbons.

The present application provides antibodies, fusion proteins and their uses for treating coronavirus. The 6-HB interfering polypeptide in the fusion protein of the present application cooperates with the antibody portion or antigen binding fragment to prevent SARS-CoV or SARS-CoV-2 virus particles from fusing with cells, and to mediate phagocytosis of immune cells, clearing virus particles.

The application discloses preparation and application of anti-Fusobacterium nucleatum PknK and PknL protein monoclonal antibodies and belongs to the technical field of biotechnology. The application takes two virulence proteins PknK and PknL of the Fusobacterium nucleatum as immunogens, successfully prepares monoclonal antibodies against the serine / threonine protein kinases PknK and PknL of the Fusobacterium nucleatum through hybridoma cell fusion technology, and the two monoclonal antibodies against the PknK and PknL proteins of the Fusobacterium nucleatum have good safety, thereby providing a basis for clinical drug safety. More importantly, the antibodies can not only efficiently neutralize bacterial toxins and directly eliminate the pathogenicity of the Fusobacterium nucleatum, but also can be directly combined on the bacterial surface to enhance the recognition and phagocytosis efficiency of immune cells to the bacteria, thereby clearing pathogenic bacteria, and is expected to be developed into a drug for resisting the Fusobacterium nucleatum infection and has a very good application prospect.

The invention discloses a monoclonal antibody, a hybridoma cell strain, a detection kit, a detection method and application, and relates to the technical field of animal epidemic disease prevention and control. According to the invention, a monoclonal antibody is obtained and is obtained by secreting a hybridoma cell strain DRV-4G3 with the preservation number of CGMCC (China General Microbiological Culture Collection Center) No.46567. The specific monoclonal antibody aiming at the duck reovirus muNS protein is prepared and obtained through an immune mouse and splenocyte fusion technology, the monoclonal antibody can recognize the muscovy duck reovirus and the novel reovirus at the same time, and the method for detecting the duck reovirus is established on the basis of the monoclonal antibody and can be used for large-scale serological detection. And reovirus vaccine immune antibody dynamic monitoring is achieved, and good development and practical application values are achieved.

The invention relates to the technical field of virus utilization, and particularly provides a method for improving lentivirus titer, which is used for screening compounds, compound concentration and harvesting time used in a production process. On one hand, the mechanism provided by the invention is helpful for reducing reinfection of produced lentivirus BaEVless on production cells and improving virus titer; on the other hand, in the production process, the heparin sodium can remarkably reduce apoptosis after production cell fusion, prolong the cell survival time and increase the virus yield.

The invention discloses an anti-Burkholderia gladioli coconut poison pathogenic variant monoclonal antibody hybridoma cell strain and a preparation method thereof, and relates to the technical field of Burkholderia gladioli. The strain is obtained by immunizing a BALB / c mouse with an antigen prepared from B. gladioli C.; the preparation method comprises the following production steps: S1, preparing an antigen; s2, immunizing the mouse; s3, carrying out cell fusion; s4, selective culture of hybridoma cells; and S5, identifying. Compared with a traditional microculture method and a molecular biological method, the antibody detection system provided by the invention has the advantages that the detection sensitivity is improved, the detection time is effectively shortened, the detection convenience is improved, and a stable basis is provided for an immunoassay method of B. gladioli C.

This invention discloses a method for preparing mesenchymal stem cells (MSCs) through synergistic pretreatment with hypoxia and cytokines, comprising: taking passaged MSCs and culturing them to a cell confluence of 70%–85%; culturing the cells under a first hypoxic condition with an oxygen volume fraction of 4%–6% for 6–18 hours; then culturing them under a second hypoxic condition with an oxygen volume fraction of 0.5%–3% for 12–36 hours; subsequently, adding a cytokine composition including IFN-γ, TNF-α, and IL-1β under the second hypoxic condition for 6–24 hours of pulsed stimulation; removing the cytokine-containing culture medium, adding a cytokine-free culture medium, and restoring the culture under an oxygen volume fraction of 2%–5% for 4–12 hours, and harvesting the pretreated MSCs. This invention improves the hypoxia tolerance, immunomodulatory capacity, and paracrine repair capacity of MSCs through a synergistic process of hypoxia adaptation, deep hypoxia induction, cytokine pulsed stimulation, and stabilization and restoration culture.

The application provides a method for efficiently expressing and producing BMP-2 in Escherichia coli. The synergistic effect of the host cell, the BMP-2 fusion protein and the high-density fermentation and induction strategy optimized in the application promotes the high-level expression of the target protein in a soluble and active form, so that the BMP-2 fusion protein accounts for more than 30% of the total soluble protein, the volume yield of the fusion protein in the fermentation liquor reaches an amazing yield of 26 g / L, and the fusion protein has activity after purification. The application simplifies the downstream purification process, reduces the production cost, and fundamentally overcomes the key bottleneck of inclusion body formation existing in the traditional method.

Vaccines and methods. Methods for identifying optimized antigenic pathogen polypeptides capable of inducing a broad neutralizing immune response to pathogens and related T cell responses are described, as well as nucleic acid sequences encoding such polypeptides. Also described are methods for determining whether a broad neutralizing immune response is induced in a subject after immunization with an optimized antigenic pathogen polypeptide or a nucleic acid encoding an optimized pathogen polypeptide. Nucleic acid molecules, polypeptides, vectors, cells, fusion proteins, pharmaceutical compositions, and their use as vaccines against pathogens, particularly against newly occurring or reoccurring pathogens, particularly RNA viruses, are also described.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody for HPV16 type E7 protein detection and application of the monoclonal antibody. E7 protein serving as an antigen and a Freund's complete adjuvant are mixed, then a mouse is subjected to primary immunization, then the E7 protein and a Freund's incomplete adjuvant are mixed, then the mouse is subjected to enhanced immunization, and the HPV16 type E7 protein detection monoclonal antibody is obtained. The monoclonal antibody for detecting the HPV16 type E7 protein is prepared by taking mouse splenocytes and myeloma cells to be fused to prepare hybridoma cells, then performing clone detection on the hybridoma cells to screen the hybridoma cells aiming at the specificity of the HPV16 type E7 protein, and finally obtaining the monoclonal antibody for detecting the HPV16 type E7 protein. According to the present invention, the HPV16 type E7 antigen detection kit is developed by using the immunochromatography method, and the kit has advantages of high sensitivity, strong specificity, simple operation and fast detection speed, and can be used for rapid screening diagnosis and illness state monitoring of HPV16.

The invention belongs to the field of stem cell biology, and relates to a culture method of a 3D neural immune organoid containing microglial cells, which comprises the following steps: S1, respectively inducing induced pluripotent stem cells in an ectodermal direction and a mesodermal direction to obtain a neural embryoid and a yolk sac embryoid; s2, culturing the neural embryoid to perform neural differentiation to obtain a neural garland structure, and re-digesting the neural garland structure to obtain a neural progenitor cell single cell; culturing the yolk sac embryoid to enable the yolk sac embryoid to generate macrophage progenitor cells; and S3, fusing the neural progenitor cell single cell and the macrophage progenitor cell, and continuously culturing to form the 3D neural immune organoid containing the microglial cells. The brain-like organ provided by the invention has the functions of exploring the control of microglial cells on the progenitor cell proportion, the change of phagocytic function, the control on the number of mature neurons and the influence on cell proliferation and apoptosis in the development stage, and meanwhile, the brain-like organ can accept external stimulation and make stress change of corresponding functions.

The invention relates to a monoclonal antibody capable of recognizing a human MGMT antigen, a preparation method of the monoclonal antibody and application of the monoclonal antibody in immunodetection. According to the technical scheme, the full-length protein is selected as the antigen peptide, codon optimization is carried out, a gene segment suitable for being expressed in escherichia coli BL21 is formed, and the finally obtained recombinant protein contains an MGMT protein and a histidine protein tag. The recombinant protein is used for immunizing a mouse, and through cell fusion, screening and subcloning, a mouse hybridoma cell strain secreting the anti-MGMT protein monoclonal antibody and the anti-MGMT protein monoclonal antibody secreted by the cell strain are obtained. The antibody obtained by the scheme has high specificity and sensitivity, can specifically recognize cells expressing MGMT protein, and is suitable for immunological detection, especially immunohistochemical detection.

The application discloses a chicken MTRF1L protein monoclonal antibody and application thereof, and belongs to the technical field of biotechnology.The MTRF1L recombinant protein is used for immunizing animals, a sensitized B cell with the ability of secreting specific antibodies and a myeloma cell with unlimited reproduction capacity are fused into a B cell hybridoma through a hybridoma technology, and a chicken MTRF1L protein monoclonal antibody is obtained through screening.The application screens a high-quality nucleotide coding sequence of the MTRF1L gene through bioinformatics analysis, and uses the nucleotide coding sequence as an antigen to immunize BABE mice, a sensitized B cell with the ability of secreting specific MTRF1L antibodies and a myeloma cell with unlimited reproduction capacity are fused into a B cell hybridoma through a hybridoma technology, and a positive hybridoma cell is obtained through screening, and the cell supernatant is used for Western Blot and indirect immunofluorescence detection, and it is found that the monoclonal antibody has good specificity and high sensitivity, which lays a foundation for detection of the MTRF1L protein and research on pathological mechanisms of virus infection diseases.

To provide a composition for promoting membrane fusion and use thereof.SOLUTION: In some aspects, fusosome compositions and methods are described herein, including membrane-encapsulated preparations comprising a fusogen. In some embodiments, the fusosome is capable of fusing with the target cell, thereby delivering the conjugated biological agent to the cytoplasm of the target cell. The present disclosure provides, in some aspects, a fusosome comprising (a) a lipid bilayer, (b) a lumen (e.g., comprising a cytosol) surrounded by the lipid bilayer, and (c) an exogenous or overexpressed fusogen, e.g., wherein the fusogen is disposed within the lipid bilayer, wherein the fusosome is derived from a source cell, and wherein the fusosome has partial or complete nuclear inactivation (e.g., removal of the nucleus).SELECTED DRAWING: Figure 1

The invention relates to a fusion type membrane structure engineering cell as well as a preparation method and application thereof. According to the method, stable expression after gene electrotransduction is achieved by combining stem cell fusion means, non-virus-dependent fusion type engineering cells can be obtained, good safety is achieved, a membrane structure treatment method is further designed, the fusion type membrane structure engineering cells are prepared, and compared with a traditional irradiation treatment mode, the method is more stable and efficient, and good application prospects are achieved. The obtained fusion type membrane structure engineering cell can be used as a feeder cell, in addition, a method for in-vitro amplification of tumor infiltration T lymphocytes and tumor infiltration NK cells is further developed, large-scale and efficient preparation of the tumor infiltration T lymphocytes and the tumor infiltration NK cells with excellent tumor killing activity can be realized by utilizing cancerous pleuroperitoneal fluid, and the method is suitable for large-scale preparation of the tumor infiltration T lymphocytes and the tumor infiltration NK cells. Technical support is provided for preparation of tumor vaccines, high-activity CAR-T and UCAR-T, high-activity CAR-NK and UCAR-NK and the like, and meanwhile a new path and a new method are provided for cancer treatment through a cell treatment method.

Bacterial antigen presenting cells are provided. Fusion proteins comprising a peptide-MHC complex, linker, and an autotransporter can be expressed on the surface of gram-negative bacteria, which can selectively stimulate T-cells reactive to the peptide-MHC complex. The bacterial antigen presenting cells can be used, e.g., to stimulate T-cells in vitro or in vivo.