47 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 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 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.

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.

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 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 invention relates to the technical field of biology, in particular to a human umbilical cord mesenchymal stem cell culture method which comprises the following steps: pretreating a fresh umbilical cord, removing blood vessels and outer membranes, flushing and cutting into pieces; separating cells through double-enzyme digestion, screening, centrifuging, resuspending by using a serum-free culture medium A, and inoculating into a coating culture bottle; performing primary culture until the cell fusion degree is 80-90%; digesting the cells during passage, and inoculating the cells to a three-dimensional scaffold containing a culture medium B in proportion; the liquid is regularly changed in the three-dimensional culture, and the culture lasts for 4-6 generations. Efficient separation and adherence of stem cells are achieved through a double-enzyme gradient digestion technology and a three-dimensional sodium alginate-gelatin porous scaffold, a serum-free culture medium A containing human platelet lysate (HPL) and the like is adopted for primary culture, the proliferation efficiency is improved, and cell activity and high dryness are maintained; in the passage stage, a culture medium B containing umbilical cord blood serum substitute (UCB-SR) and the like is used, a low-oxygen environment and triaxial tensile stress stimulation are combined, apoptosis is inhibited, and the purity and dryness of stem cells are ensured.

Provided are a cow milk-derived stem cell, and an isolation and culture method therefor. The method comprises the following steps: (1) taking fresh cow milk, and isolating cells therefrom; (2) coating a culture dish with a matrix gel in advance, discarding the coating solution, then washing the culture dish with a washing solution, and discarding the washing solution; (3) inoculating the cells treated in the step (1) on the coated culture dish, and culturing the cells with a stem cell culture medium; and (4) when the cell confluence reaches 80% or more, digesting the cells with a digestion enzyme, terminating the digestion, isolating the cells, resuspending the cells using a CTFR culture solution, then inoculating the cells on a feeder layer for culture, promoting colony formation, and performing subculture to obtain cow milk-derived stem cells. The cow milk-derived stem cell has OCT4, SOX2, NANOG and SSEA4 markers.

The application provides a kind of sandwich detection testosterone monoclonal antibody and its preparation method and application, comprising: step S1, preparation testosterone-3-(O-carboxymethyl) oxime, and it is activated;After activation, take testosterone antibody P2G1, according to proportion, drop the activated testosterone-3-(O-carboxymethyl) oxime is reacted to prepare immunogen;Step S2, based on the prepared immunogen, immunization animal;Step S3, the spleen cell of immunization animal and myeloma cell are fused into hybridoma cell, and the real hybridoma cell is selected;Step S4, antibody is separated and purified from the screened hybridoma cell.The application provides a kind of preparation method of raw material and reagent for testosterone sandwich method detection, solves the problem that the sensitivity of small molecule compound testosterone competition method reagent is insufficient, detection range is narrow, specificity is poor, provides a new thought and method for the rapid, accurate detection of testosterone.

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 present invention discloses a method for constructing an Alzheimer's disease cell model, comprising the following steps: (1) preparing senecionine: under sterile conditions, weighing senecionine powder, fully dissolving it with a 5% sterile hydrochloric acid aqueous solution by vortexing, adding 1mM NaOH to adjust the pH to 6-7, and preparing a 500mM senecionine stock solution; (2) cell culture: SH-SY5Y cells were cultured in MEM / F-12 complete medium in a 5% CO2, 37°C incubator, and the complete medium was 44% MEM+44% F-12+10% FBS+1% P / S+1% Glutamine; (3) Cell treatment: SH-SY5Y cells in the logarithmic growth phase were taken and the cell density was adjusted to 50,000 cells per well. They were seeded in a 6-well plate and placed in an incubator. 100 μL of 42-84 μM Senecionine solution was added to each well. When the cell confluence reached 58%-60%, the solution was replaced and the culture was continued for 48 hours. This invention is the first to use Senecionine to construct a cell model of Alzheimer's disease (AD). The cells used were SH-SY5Y and the Senecionine concentration was 42-84 μM. After 48 hours of culture, a cell model of AD was successfully constructed.

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.

Methods, and compositions of matter useful for enhancing activity of endogenous and / or exogenous regenerative cells by selectively eliminating senescent cells through induction of immunity against said senescent cells or components thereof. Fusion of autologous patient cells made senescent, autologous antigen presenting cells and utilized as a vaccine. Autologous fibroblasts can be made senescent by genotoxic stress and fused with dendritic cells. Said dendritic cells may be generated from circulating monocytes, CD34 cells or autologous iPSC cells.

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.