83 results about "Cell fusion" patented technology

The invention provides a monoclonal antibody capable of recognizing a recombinant EGF-CRM197 vaccine and a preparation method thereof, on the basis of researching, developing and preparing the recombinant EGF-CRM197 vaccine, after the recombinant EGF-CRM197 vaccine is further immunized and inoculated to a mouse, mouse spleen cells and mouse myeloma cells are fused, and the monoclonal antibody capable of recognizing the recombinant EGF-CRM197 vaccine is obtained. And screening a hybridoma cell strain which generates a specific monoclonal antibody for recognizing the recombinant EGF-CRM197 vaccine, and sequencing to obtain an antibody sequence. After the hybridoma cells are cultured in a mouse body, the monoclonal antibody for recognizing the recombinant EGF-CRM197 vaccine can be prepared in vitro through methods of chromatographic purification and the like. The method can be used for detecting the release titer of the recombinant EGF-CRM197 vaccine and evaluating the immune efficacy after the recombinant EGF-CRM197 vaccine is immunized.

The invention discloses an anti-RAGE antibody, and also discloses a conjugate containing the anti-RAGE antibody, a bispecific antibody, a multispecific antibody, an immune cell, a fusion protein and the like. The anti-RAGE antibody provided by the invention can target RAGE antigen targets of different species of human, mouse and monkey, and has very high application value in the aspects of scientific research and clinical application.

The invention discloses an NK cell culture method with high amplification efficiency in the technical field of NK cells, by constructing a recombinant plasmid pGAPZa-NDUFA8-NDUFB8 and realizing high-efficiency expression and purification of a fusion protein on the basis, the amplification efficiency of the NK cells can be remarkably improved, and by introducing the NDUFA8-NDUFB8 fusion protein, the amplification efficiency of the NK cells is remarkably improved. A sufficient number of active NK cells can be obtained in a short time, and the fusion protein not only promotes cell proliferation, but also maintains high killing ability and immunocompetence of the amplified NK cells.

The invention discloses a dental sac mesenchymal stem cell with high periostin expression as well as an extraction and culture method and application thereof, and belongs to the technical field of biomedicine. The dental sac mesenchymal stem cell with high periostin expression is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C2024384; the extraction and culture method disclosed by the invention comprises the following steps: S1, extraction: cleaning, cutting, digesting and centrifuging dental sac tissues, resuspending, centrifuging and precipitating by using a primary culture medium, culturing until cells are fused, digesting, and carrying out secondary centrifugation and precipitating to obtain primary dental sac mesenchymal stem cells; s2, subculture: resuspending the dental sac mesenchymal stem cells in the step S1 by using a primary culture medium, supplementing a subculture medium, and then subculturing. The invention also discloses the dental sac mesenchymal stem cell with high periostin expression and application of the dental sac mesenchymal stem cell in preparation of medicines for preventing or / and treating periodontal diseases. The method disclosed by the invention has an unexpected effect of enhancing periostin expression of the dental sac mesenchymal stem cells.

Described are lipopeptides that inhibit coronavirus fusion to a host cell. Thus a therapeutic for treating or preventing the common cold is described along with methods of inhibiting and / or treating an alphacoronavirus infection. The lipopeptides comprise a peptide unit comprising an amino acid sequence having a high degree of sequence identity to a sequence from the C-terminal heptad repeat of a betacoronavirus S protein, such as that of SARS-CoV-2.

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.

The invention belongs to the technical field of biological carriers, and particularly relates to a preparation method and application of an exosome ferulic acid nano-composite. The invention provides a way for preparing ferulic acid nano-particles by using exosomes, and particularly relates to a preparation process for forming a nano-composite by using ferulic acid and stress yeast exosomes generated by stimulation. The preparation method of the exosome ferulic acid nano-composite provided by the invention has the advantages of high safety, low immunological rejection, high cell fusion degree, high entrapment rate and good stability. Besides, the selected stress yeast exosome can effectively protect and enhance the anti-inflammatory and anti-oxidation effects of ferulic acid, so that the prepared exosome ferulic acid nano-composite has relatively strong anti-inflammatory, anti-oxidation and repair effects, and has a good application prospect in the field of cosmetics or medicines.

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 invention discloses an anti-phenacetin monoclonal antibody hybridoma cell strain AD, a preparation method and application thereof, and relates to the technical field of food safety immunodetection. The monoclonal antibody hybridoma cell strain is named monoclonal cell strain AD and the number CGMCC19681. The Phe-BA obtained by the hydrolysis of the reaction product of the phenacetin metabolite acetaminophen and ethyl 4-bromobutyrate is used as the hapten, and the hapten is coupled with the carrier protein to prepare the immunogen Phe-BA-BSA. After the mice were immunized with the immunogen Phe-BA-BSA, they were fused with myeloma cells by PEG method, screened by indirect competitive enzyme-linked immunosorbent assay and subcloned five times to obtain hybridoma cell lines. The monoclonal antibody secreted by the cell line can be made into a phenacetin detection kit, which has good affinity and detection sensitivity for phenacetin, and can be used for immunodetection of phenacetin residues in food.

The present invention discloses an in vitro preparation method of human mesenchymal stem cells. The method comprises the following steps: using MSC culture medium supplemented with chemical small molecules to induce iPSC-derived MSCs; after the iPSC cell confluence reaches 40%, the culture medium containing chemical small molecules is replaced; after 8-10 days of induction, the cells are collected and passaged; and the cells are cultured to the P1 generation, showing uniform fibroblast-like, spindle-shaped or fusiform cells. The prepared hiPSC-MSCs meet the relevant phenotypes of mesenchymal stem cells, wherein the positive rates of CD73, CD90 and CD105 are higher than 95%, and the cells have immunomodulatory activity.

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 invention discloses a mutant streptococcus protein G. The invention also discloses a polynucleotide, an expression vector, a host cell and a fusion protein. The invention also discloses an adsorption material, wherein the mutant streptococcus protein G or the fusion protein is fixed on a carrier to obtain the adsorption material. The invention also discloses application of the mutant streptococcus protein G, the fusion protein and the adsorption material in separation and purification of F (ab ') 2 fragments and derivatives thereof as chromatographic column packing. The invention also discloses a method for separating and purifying the F (ab ') 2 fragment, which comprises the following steps: carrying out column chromatography on a sample containing the F (ab') 2 fragment by using the adsorption material as a chromatographic column filler to obtain the F (ab ') 2 fragment. The mutant streptococcus protein G can specifically target an F (ab ') 2 fragment, is not combined with the performance of an Fc fragment, and can quickly and efficiently purify the F (ab') 2 fragment from horse serum.

The present application discloses an anti-RAGE antibody, and also discloses a conjugate, bispecific, multispecific antibody, immune cell, fusion protein, etc. comprising the anti-RAGE antibody. The anti-RAGE antibody of the present application can target RAGE antigen targets of different species such as human, mouse and monkey, and has high application value in scientific research and clinical application.

The present application relates to a kind of monoclonal antibody that can identify human Topo II alpha antigen, secretes cell strain, its preparation method and its use in immunodetection.The above technical solution selects the amino acid of 1301-1531 of Topo II alpha protein as antigen peptide, codon optimization becomes the gene fragment suitable for expression in escherichia coli BL21, and finally the recombinant protein includes Topo II alpha protein fragment and histidine protein label.The recombinant protein is immunized to mouse, is fused by cell, screening and subcloning, and obtains mouse hybridoma cell strain, and the anti Topo II alpha protein monoclonal antibody secreted by the cell strain is obtained.The antibody obtained by the present application has high specificity, sensitivity, and can specifically recognize the cell expressing Topo II alpha protein, and is suitable for immunological detection, especially immunohistochemical detection.

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 relates to a preparation method and application of hybridoma cells based on an electrofusion technology, the preparation method of the hybridoma cells based on the electrofusion technology comprises the following steps: taking splenocytes and myeloma cells of an antigen-immunized mouse for electrofusion, screening to obtain the hybridoma cells, and the antigen is serum amyloid protein A. The invention also relates to a preparation method of the hybridoma cells based on the electrofusion technology. The hybridoma cell capable of generating the serum amyloid protein A antibody is obtained in an electrofusion mode, the fusion efficiency of the hybridoma cell is improved, and the problem that the yield of the antibody is reduced due to the fact that the fusion efficiency of the hybridoma cell is low is solved.

To provide vaccines and methods.SOLUTION: Described herein are methods for identifying optimized antigenic pathogen polypeptides capable of inducing a broadly neutralizing immune response and associated T-cell responses to a pathogen, and nucleic acid sequences encoding such polypeptides. Also described are methods for determining whether a broadly neutralizing immune response is induced in a subject following immunization with an optimized antigenic pathogen polypeptide or a nucleic acid encoding the optimized pathogen polypeptide. Further described are nucleic acid molecules, polypeptides, vectors, cells, fusion proteins, pharmaceutical compositions, and their use as vaccines against pathogens, especially against emerging or re-emerging pathogens (particularly RNA viruses).SELECTED DRAWING: None

The invention discloses a bionic photo-thermal nano motor for targeted regulation and control of a solid tumor matrix-immunosuppression microenvironment and a preparation method and application thereof, and belongs to the technical field of biomedical materials.The preparation method comprises the steps that polydopamine nanoparticles and a K2PtCl4 aqueous solution are subjected to condensation reflux, and asymmetric PDA-Pt is prepared; and preparing a cancer-related fibroblast and breast cancer cell fusion membrane and coating the surface of PDA-Pt with the cancer-related fibroblast and breast cancer cell fusion membrane. The nano-motor realizes dual targeting of matrix cells and tumor cells through a surface modified fusion cell membrane, targeted ablation CAFs regulates and controls a tumor matrix microenvironment, a good tumor permeation effect is realized by cooperating with H2O2 responsive self-driving of the nano-motor, a mild photo-thermal-catalytic synergistic effect is formed by a photo-thermal effect of PDA and peroxidase-like activity of Pt nano-enzyme, and a good tumor permeation effect is achieved. And finally, the tumor immunosuppression microenvironment is remarkably improved, and the tumor immunotherapy effect is improved.

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.

The present application discloses an anti-RAGE antibody, and also discloses a conjugate, bispecific, multispecific antibody, immune cell, fusion protein, etc. comprising the anti-RAGE antibody. The anti-RAGE antibody of the present application can target RAGE antigen targets of different species such as human, mouse and monkey, and has high application value in scientific research and clinical application.

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 invention belongs to the technical field of biological medicine, and particularly discloses a complex tumor organoid co-culture system and a construction method and application thereof. The construction method comprises the following steps: construction of tumor organs, separation and induced differentiation culture of dendritic cells, fusion of tumor cells and maturely differentiated dendritic cells (DCs) through oncolytic viruses, and addition of immune cells for co-culture. According to the present invention, the patient-derived tumor cells are utilized to construct the tumor organoid, such that the pathological characteristics of the patient can be well restored; the oncolytic virus is used for inducing fusion of the dendritic cells and the primary tumor cells, so that related antigens, specific antigens and new antigens of the tumor cells can be presented more effectively, and the reaction of immune cells can be stimulated more efficiently; the rapid, simple and effective complex tumor organoid co-culture established by the invention can simulate the positioning and interaction relationship between immune cells and tumor cells in an in-vivo tumor microenvironment, and also provides a platform for evaluation of a novel immunotherapy.

The invention discloses a production method of mesenchymal stem cells applied to anti-aging, and relates to the technical field of cell biologication.The production method of the mesenchymal stem cells applied to anti-aging comprises the following steps that S1, cell culture is conducted, specifically, mesenchymal stem cells are cultured in vitro till the cell fusion degree reaches 60%-95%; s2, cytokine stimulation: adding cytokines into the cultured cell culture medium, and stimulating for 24-48 hours; s3, detecting and screening: digesting and collecting the mesenchymal stem cells, detecting the expression level of indoleamine 2, 3-dioxide oxidase and related cytokines in the mesenchymal stem cells, and detecting the immune regulation ability of T cells; the mesenchymal stem cells obtained by adopting the production method of the mesenchymal stem cells applied to anti-aging provided by the invention have a better anti-aging effect, the immune regulation ability and the specific factor secretion level of the mesenchymal stem cells are improved by adding cytokines, the production method of the mesenchymal stem cells is established, and the mesenchymal stem cells can be used for preparing the anti-aging mesenchymal stem cells. The pharmaceutical composition is applied to anti-aging related diseases.

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.

The invention discloses a preparation method of hypertonic stimulated bone marrow mesenchymal stem cells, and particularly relates to the technical field of stem cell culture. The method comprises the following steps: S1, adding a sodium chloride solution into a DMEM / f-12 complete medium to prepare a hypertonic medium system; s2, selecting third-generation bone marrow mesenchymal stem cells, and replacing the hypertonic culture medium when the cells are fused to 70-80%; s3, after the culture medium is replaced, culturing in a constant-temperature cell culture box to obtain the bone marrow mesenchymal stem cells subjected to hypertonic stimulation treatment. The method can effectively improve the cell activity and multiplication capacity of the bone marrow mesenchymal stem cells, delay cell senescence and enhance the anti-active oxygen and anti-hypertonic capacity, thereby improving the treatment effect on the radiation-induced lung injury.

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.