156 results about "Cell-cell fusion" patented technology

The invention discloses a micro-fluidic chip device for cell fusion and in-situ sorting. The micro-fluidic chip device comprises a PDMS (Polydimethylsiloxane) micro-fluidic chip, a PCB (Printed Circuit Board) and a patterned gold electrode micro-array layer which are sequentially arranged from top to bottom, the PDMS micro-fluidic chip is provided with a sample inlet, a sample outlet and a collection cavity, and the sample inlet, the sample outlet and the collection cavity are communicated through the experiment cavity; a plurality of independent channels and a capturing cavity connected with the independent channels are arranged in the experiment cavity, and a double-cavity capturing structure is arranged at an inlet of the capturing cavity; and an independent microelectrode matched with the double-cavity capture structure is arranged on the patterned gold electrode microarray. The independent microelectrodes on the patterned gold electrode microarray layer are matched with the double-cavity capture structure, so that cell capture, cell electrofusion and cell in-situ release can be realized in the same chip, and cells in the double-cavity capture structure can be controlled more accurately.

The invention belongs to the technical field of biology, and particularly relates to a latex duplex immunochromatography test strip for rapidly identifying a porcine transmissible gastroenteritis virus (TGEV) and a porcine epidemic diarrhea virus (PEDV) and a preparation method of the latex duplex immunochromatography test strip. Soluble TGEV N protein and PEDV N protein with immunogenicity and reactivity are respectively obtained by utilizing a prokaryotic expression system, the soluble TGEV N protein and the soluble PEDV N protein are used as immunogenicity to immunize mice, and specific monoclonal antibodies of the TGEV N protein and the PEDV N protein are respectively obtained by utilizing cell fusion and subcloning screening technologies. The antibody variable region gene sequence is obtained by using a nested PCR amplification technology, and can be subsequently obtained by using a gene engineering or protein engineering method. Then, the latex duplex immunochromatography test strip for detecting TGEV and PEDV, which is convenient, rapid and high in sensitivity, and the preparation method are researched and developed on the basis of specific monoclonal antibodies of TGEV N protein and PEDV N protein.

The invention is applicable to the technical field of C-reactive protein detection, and provides an anti-C-reactive protein monoclonal antibody idiotype antibody, a preparation method and application, and the anti-C-reactive protein monoclonal antibody idiotype antibody is secreted by a hybridoma cell strain; the hybridoma cell strain is a 7C4 hybridoma cell strain; the preparation method of the anti-C-reactive protein monoclonal antibody idiotype antibody comprises the following steps: S1, obtaining hybridoma cells: preparing an anti-C-reactive protein monoclonal antibody Fab fragment, then carrying out animal immunization, then carrying out cell fusion, and finally screening and cloning the hybridoma cells; and S2, preparing and screening an anti-C reactive protein monoclonal antibody idiotype antibody. The anti-C-reactive-protein monoclonal antibody idiotype antibody is used for replacing C-reactive protein to serve as a labeled antibody, the labeling process is simple, the labeling efficiency is high, and meanwhile the situation that a final product is affected due to instability of the C-reactive protein is avoided. Experimental results show that the detection method is good in specificity and wide in linear range.

A specific protein fragment capable of representing salmonella as an antigen is obtained through screening and experimental verification, a mouse is immunized with the specific protein fragment as the antigen, and hybridoma cells capable of stably secreting monoclonal antibodies are obtained through a cell fusion technology. And finally, the anti-salmonella monoclonal antibody with high titer and good specificity is prepared. Similarly, the specific protein fragment is used for immunizing a rabbit to obtain the anti-salmonella polyclonal antibody. On the basis of a mouse monoclonal antibody and a rabbit polyclonal antibody, a salmonella colloidal gold immunochromatography detection method is established, the detection method is good in specificity, and a rapid, simple and convenient detection means is provided for detection of salmonella in food.

The invention discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody generated by the herbicide-resistant protein GAT hybridoma cell strain and an application of the herbicide-resistant protein GAT hybridoma cell strain. The herbicide-resistant protein GAT hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46333. The preparation method comprises the following steps: a) purifying prokaryotic expression to obtain GAT recombinant protein; b) immunizing animals: immunizing BALB / c mice by taking the GAT recombinant protein as an antigen; c) cell fusion: splenocytes of the immune BALB / c mice are collected and fused with SP2 / 0 cells; and d) cell strain establishment: carrying out subcloning through a limited dilution method, carrying out ELISA detection after 5-7 days of subcloning until a hybridoma cell strain capable of stably secreting a positive antibody is screened out, and carrying out amplification, re-culture and storage. The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for detection of herbicide-resistant protein GAT in transgenic crops, so that development, seed production and seed conservation of transgenic gat crops are supported, and stable industrial promotion of the transgenic gat crops and sustainable application of related transformants are guaranteed.

The invention belongs to the technical field of biology, provides an anti-IL-8 monoclonal antibody, a variable region sequence of the anti-IL-8 monoclonal antibody and application of the anti-IL-8 monoclonal antibody, and discloses a monoclonal antibody with neutralizing interleukin-8 (IL-8) biological activity and the variable region sequence of the monoclonal antibody. The preparation method comprises the following steps: by taking IL-8 as an immune source and a mouse as an immune object, carrying out immune injection on the mouse, taking the spleen of the immune mouse to prepare a suspension, and carrying out cell fusion on the suspension and myeloma cells to obtain a hybridoma cell strain capable of expressing the anti-IL-8 monoclonal antibody so as to prepare the anti-IL-8 monoclonal antibody; according to the invention, a variable region amino acid sequence of the anti-IL-8 monoclonal antibody is cloned at the same time; the anti-IL-8 monoclonal antibody disclosed by the invention can be specifically combined with IL-8, so that the anti-IL-8 monoclonal antibody can be used for treating diseases related to abnormal or excessive or out-of-control expression and secretion of the IL-8.

Disclosed herein include methods, compositions, and kits suitable for use in e.g., nucleic acid delivery. Provided are compositions (e.g., nucleic acid compositions) comprising polynucleotide(s) encoding a fusion protein, a dimerization fusion protein, a soluble RBP, an adapter fusion protein, and / or a cell fusion protein. The compositions also comprise one or more polynucleotides comprising one or more RNA cargo molecules. The fusion protein, dimerization fusion protein, and / or adapter fusion protein can comprise an endosomal sorting complex required for transport (ESCRT)-recruiting domain (ERD). Recruitment of one or more ESCRT proteins results in secretion of enveloped nanoparticles (ENPs) from a cell in which at least one of a fusion protein, a dimerization fusion protein, and / or an adapter fusion protein are expressed. The ENPs can comprise the one or more RNA cargo molecules. There are also provided populations of ENPs in some embodiments.

The invention relates to the technical field of plant cell engineering, and particularly discloses a plant protoplast semi-fusion breeding method which comprises the following three stages of seedling material selection, preparation, fusion, induction and cultivation. A large number of fused plant seedlings are obtained through a series of operations and used for large-scale planting. The method comprises the following steps: selecting excellent varieties of a plant A and a plant B, respectively preparing the plant A and the plant B into protoplasts in a preparation process, culturing the protoplasts into corresponding calluses, and performing half-cell fusion on a fusion part to obtain mosaic calluses. Then, the callus of the plant A to be used as the root is induced to locally induce adventitious roots, the callus of the plant B to locally induce adventitious buds, and finally, breeding is carried out to obtain the super seedlings. According to the method, the grafting principle is combined, the degree and conditions of cell fusion are optimized, the breeding cost is reduced, meanwhile, it is ensured that the fusion body can stably express the excellent characters of parents at different parts, and agricultural large-scale production is facilitated.

The invention provides a collaborative drug prediction method and system based on a heterogeneous hypergraph, and relates to the field of collaborative drug prediction.The collaborative drug prediction method comprises the steps that information of various cells and structures of various drugs are obtained, and corresponding cell fusion features and drug structure features are obtained; constructing a heterogeneous hypergraph based on the drug structure features and the cell fusion features, performing convolution processing to obtain drug node features, cell node features and hyperedge features, and training a neural network by using the extracted features to obtain a prediction model; and obtaining a to-be-detected drug, and inputting the to-be-detected drug into the prediction model to obtain a prediction result. According to the method, the heterogeneous hypergraph is constructed based on the collaborative information of the known drug combination in the cells, and the feature information of the drug nodes, the cell nodes and the hyperedges is extracted by adopting the difference of the multilayer hypergraph convolutional layers, so that the defects of excessive smoothness and redundancy of the information during feature extraction of the heterogeneous network by a common graph neural network are overcome; and a complex relation mode is difficult to capture.

The invention relates to a multifunctional integrated organoid culture device. The device comprises a main body, and a liquid input module, a temperature control module and an organoid module which are fixed on the main body, the main body comprises an adjusting assembly and a main control chip which are connected with each other, and the main control chip is connected with the temperature control module and the liquid input module and used for adjusting the flow speed of the liquid input module and the temperature of the temperature control module; the liquid input module is connected with the organ-like culture module and used for conveying cell sap to the organ-like culture module, the organ-like module is provided with a cell anchoring array and used for fixing and culturing cells, and the temperature control module is located between the liquid input module and the organ-like module and used for controlling the temperature of the organ-like culture module. The temperature detection module is used for detecting the temperature of the liquid input module and adjusting the temperature of the organ-like module. Compared with the prior art, the invention has the advantages of high control precision, multi-cell fusion, abundant functions and the like.

The invention discloses an amniotic mesenchymal stem cell exosome and a preparation method and application thereof.The preparation method comprises the following steps that amniotic mesenchymal stem cells are inoculated into a complete culture medium to be cultured, and after the cells adhere to the wall, epidermal growth factors and polypeptide are added into the complete culture medium to continue to be cultured; replacing the complete culture medium, continuously culturing until the cell fusion degree is not lower than 85%, replacing the complete culture medium again, culturing, and collecting supernate; and extracting the exosome in the supernate by adopting an ultracentrifugation method to obtain the amniotic mesenchymal stem cell exosome. According to the preparation method of the amniotic mesenchymal stem cell exosome, the epidermal growth factor and the polypeptide are added into the culture medium, so that secretion of the cell exosome can be stimulated, meanwhile, carrying substances of the exosome are improved, the functions of promoting cell proliferation, migration and the like of the exosome are improved, and then healing of skin injury can be better promoted; the wound healing time is obviously shortened, and the pain of a patient is relieved.

The application belongs to the technical field of biology and particularly relates to an African swine fever virus pNP868R protein monoclonal antibody and a preparation method and application thereof. The application takes the pNP868R protein prepared by eukaryotic expression as an antigen, immunizes BALB / c mice, uses hybridoma technology, and obtains a hybridoma cell line capable of stably secreting the anti-African swine fever virus pNP868R protein monoclonal antibody through cell fusion, indirect ELISA method screening, limited dilution method subcloning and hybridoma continuous passage culture. The application also obtains an African swine fever virus pNP868R protein monoclonal antibody. The monoclonal antibody has high affinity with the pNP868R protein and can be applied in Western-Blot experiments, indirect immunofluorescence experiments, immunoprecipitation experiments and the capture of proteins interacting with pNP868R, and can be used for the diagnosis, prevention or treatment of African swine fever.

The invention discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody generated by the herbicide-resistant protein GAT hybridoma cell strain and application of the herbicide-resistant protein GAT hybridoma cell strain and the antibody. The herbicide-resistant protein GAT hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC No.46332. The preparation method comprises the following steps: a) purifying prokaryotic expression to obtain GAT recombinant protein; b) immunizing animals: immunizing BALB / c mice by taking the GAT recombinant protein as an antigen; c) cell fusion: splenocytes of the immune BALB / c mice are collected and fused with SP2 / 0 cells; and d) cell strain establishment: carrying out subcloning through a limited dilution method, carrying out ELISA detection after 5-7 days of subcloning until a hybridoma cell strain capable of stably secreting a positive antibody is screened out, and carrying out amplification, re-culture and storage. The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for detection of herbicide-resistant protein GAT in transgenic crops, so that development, seed production and seed conservation of transgenic gat crops are supported, and stable industrial promotion of the transgenic gat crops and sustainable application of related transformants are guaranteed.

Disclosed herein are compositions that block entry of enveloped viruses into cells, polynucleotides encoding the same, pharmaceutical compositions, and methods of treating viral infections in subjects using the disclosed compositions. Also disclosed are compositions that make a cell susceptible to viral infection, polynucleotides encoding the same and methods of making a cell susceptible to viral infection, methods of targeting an oncolytic virus to a cell, and methods of inducing cell-cell fusion.

Disclosed are compositions and methods for estimating the number of cell fusion events occurring in liquid cultures using multiplexed oligonucleotide molecular barcodes and next generation sequencing.

The invention relates to a hybridoma cell strain secreting a rofecoxib monoclonal antibody and application of the hybridoma cell strain, and belongs to the technical field of immunodetection. According to the invention, a hybridoma cell strain secreting the rofecoxib monoclonal antibody is obtained through screening by preparing a rofecoxib complete antigen and carrying out animal immune reaction and cell fusion steps, the hybridoma cell strain is preserved in China General Microbiological Culture Collection Center (CGMCC), and the preservation number is CGMCC NO.46734. The monoclonal antibody secreted by the hybridoma cell strain has good sensitivity and specificity, the IC50 value to rofecoxib is 20.0 ng / mL, the cross reaction rate to various rofecoxib functional analogues is smaller than 1%, the monoclonal antibody can be applied to preparation of rofecoxib detection products, and an efficient detection method and means are provided for detection of rofecoxib residues in food.

Antiviral peptides and formulations thereof are described for use in treating or preventing one or more symptoms of coronavirus infections. Peptides derived from human beta defensin 2 have been shown to have antiviral properties against different variants of coronavirus including cross-linking viral particles, blocking cell-to-cell fusion, and / or inhibiting viral release. Pharmaceutical compositions and methods of using one or more antiviral peptides are also provided. Preferably, the antiviral peptides are administered via intranasal route to prevent or alleviate one or more symptoms of coronavirus infections such as reducing the syncytial formation and lung damage.

The invention discloses a feline leukemia virus p27 protein monoclonal antibody, a hybridoma cell strain and application thereof, and relates to the technical field of biology, the amino acid sequence of a heavy chain variable region of the monoclonal antibody is as shown in SEQ ID NO.1 in a sequence table, and the amino acid sequence of a light chain variable region of the monoclonal antibody is as shown in SEQ ID NO.2 in the sequence table. According to the invention, a cell fusion technology is utilized to establish a hybridoma cell strain p27-3A5 capable of secreting a p27 protein monoclonal antibody, and a specific monoclonal antibody 3A5 is obtained. And the minimum linear epitope identified by the antibody is identified, so that the establishment of subsequent preparation of a diagnostic kit such as a colloidal gold test strip is facilitated.

This invention belongs to the field of immunology and discloses a hybridoma cell line and the preparation and application of its secreted monoclonal antibody that uniformly recognizes tylosin / tilmicosin. The monoclonal antibody hybridoma cell line is named DES-AOAA-14D5, with accession number CGMCC NO: 45303. This invention utilizes decarboxymethyl tylosin and oxycarboxymethyl hydroxylamine to synthesize a hapten, conjugates a carrier protein to prepare a complete antigen, immunizes mice, and, through cell fusion and screening techniques, prepares a hybridoma cell line that can secrete a monoclonal antibody that uniformly recognizes tylosin / tilmicosin. IC50 50 The effective concentrations were 1.59 and 1.72 ng / mL, respectively, with a cross-reactivity rate as high as 92.44%. An immunoassay method for the simultaneous and accurate detection of tylosin / tilmicosin in milk was established using this antibody. The cut-off value for both tylosin and tilmicosin was 16 ng / mL, and no cross-reactivity was observed with other structural analogs. The detection method of this invention shows promising application prospects.

The disclosure provides the structure of an MFSD2A-SYNC2 complex together with functional data that revealed important molecular aspects of MFSD2A transport cycle, receptor-mediated cell-cell fusion, and pharmacology and resulted in the identification of two novel allosteric modulators of MFSD2A, which are two soluble fragments of SYNC2, namely SYNC2su-co and SYNC2su-co-2, representing first-in-class molecules to inhibit MFSD2A LPCs uptake and increase transcytosis rate.

The invention discloses application of a gp130 inhibitor in preparation of a medicine for treating virus infection, and relates to the technical field of medicines. Experiments prove that combination of IL-6 family cell factors and a gp130 receptor is inhibited, so that downward signal transmission of a JAK2 / STAT3 signal channel triggered by the IL-6 family cell factors is inhibited, the purposes of inhibiting cell fusion induced by SARS-CoV-2 spike protein and further influencing replication and propagation of SARS-CoV-2 are achieved, and a small-molecule inhibitor SC144 is an inhibitor of gp130, has a remarkable effect of inhibiting SARS-CoV-2 virus replication and has a good application prospect. Meanwhile, the traditional Chinese medicine composition also has a remarkable effect of inhibiting replication of viruses such as human immunodeficiency virus, herpes simplex virus and respiratory syncytial virus.

This invention provides a vaccinia virus that induces cell fusion between infected cells and a method for producing the same. Such vaccinia virus is deprived of the K2L gene or the HA gene or functions of the K2L gene and the HA gene and is mutated to induce cell fusion between infected cells and induce cell death.

The present invention discloses an intraocular irrigation solution containing niacinamide. Niacinamide serves as an active substance in the intraocular irrigation solution, which has the effects of improving the function of corneal endothelial cells, inhibiting cell apoptosis and cell fusion, and improving the mitochondrial damage condition, thereby alleviating or inhibiting corneal edema, reduction of corneal endothelial cells, abnormal corneal transparency, abnormal corneal thickness, abnormal corneal endothelial cell density, abnormal corneal endothelial cell morphology, and abnormal expression of corneal endothelial cell functional proteins caused by ultrasonic stimulation, heat stimulation or oxidative damage to corneal endothelial cells during phacoemulsification surgery.

The invention belongs to the technical field of bioengineering, and particularly relates to a preparation method and application of an anti-EGFR monoclonal antibody based on electrofusion, and the preparation method comprises the following steps: aiming at an EGFR specific effective fragment, designing an artificial antigen, and utilizing a series of processes of protein expression, mouse immunization and antibody preparation to obtain a high-specificity anti-EGFR antibody. According to the invention, cell fusion is carried out through an improved electrofusion mode, the contact rate of heterologous cells is enhanced, and the fusion rate of the cells and the number of later positive cell strains are greatly improved, so that the anti-EGFR monoclonal antibody with specificity and sensitivity both meeting the follow-up production and scientific research work is obtained; a new thought is provided for preparing medicines for preventing and diagnosing human diseases and positioning tumors and antibodies in vivo.

An object of the present invention is to provide an efficient method of cell fusion, and to provide a method of killing cancer cells by cell fusion, or a method for fusing virus envelopes by membrane fusion.The object can be solved by (1) a polypeptide comprising an amino acid sequence having a group represented by the following formula (I) on a N-terminus of an amino acid sequence selected from the group consisting of the amino acid sequences of SEQ ID NOs: 1 to 8;—Z—Xm—Y  (I)wherein Z is a hydrophilic linker, X is a hydrophilic amino acid residue selected from the group consisting of serine, threonine, asparagine, glutamine, arginine, histidine, lysine, aspartic acid, glutamic acid, tyrosine, and cysteine, Y is a carboxyl group or amino group, m is an integer of 1 to 5, when m is 2 to 5, the hydrophilic amino acid residues may be the same amino acid residue or may be a combination of different amino acid residues, or (2) a polypeptide comprising an amino acid sequence having a group represented by the above formula (I) on a N-terminus of an amino acid sequence, in which 1 to 4 amino acids are deleted, substituted, inserted, and / or added in an amino acid sequence of SEQ ID NOs: 1 to 8, and having a cell fusion activity.

The application discloses a single-chain antibody, the amino acid sequence of which is shown as SEQ ID NO:1, and the nucleotide sequence of which is shown as SEQ ID NO:2. The single-chain antibody against rabies virus is screened from an antibody library of immunized mice by using a phage display technology. Compared with a traditional cell fusion technology, the phage display technology is simple in operation, low in cost, short in time, and high in antibody discovery and screening efficiency. The single-chain antibody obtained by the application is low in molecular weight and immunogenicity, high in antibody specificity and affinity, and has the activity of neutralizing rabies virus, thereby providing a new idea for basic research of rabies virus and development of therapeutic drugs.

The invention belongs to the technical field of cell cryopreservation, and particularly relates to a cell cryopreservation solution, application thereof and a cell cryopreservation method. A death inhibitor and a solvent are selected for specific cells, and the cell death process related to cytolytic organelle membrane damage is intervened from the molecular level, specifically, the solvent is used for reducing ice crystal formation inside and outside the cells, the membrane protection effect is exerted to achieve the high resuscitation motility rate, a programmed death pathway is further blocked in a targeted mode through the death inhibitor, and the cell death effect is improved. The programmed death after cell resuscitation is relieved, the delayed death rate of the resuscitated cells is reduced, the long-term cell fusion degree of the resuscitated cells is improved, and the long-term survival rate of the resuscitated cells is improved.

The present application relates to a hybridoma cell strain secreting benzyl chlorophenol monoclonal antibody and its application, and belongs to the technical field of immune detection. The present application synthesizes benzyl chlorophenol hapten, prepares benzyl chlorophenol complete antigen, and through the steps of mouse immune response and cell fusion, a hybridoma cell strain secreting benzyl chlorophenol monoclonal antibody is screened and named CBP-3D4, which is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC NO.46222. The monoclonal antibody secreted by the hybridoma cell strain has good sensitivity and specificity, the IC 50 value of the monoclonal antibody secreted by the hybridoma cell strain for benzyl chlorophenol is 97.741 ng / mL, the cross reactivity of the monoclonal antibody for various benzyl chlorophenol functional analogs is less than 1%, and the monoclonal antibody can be applied to the preparation of benzyl chlorophenol detection products to realize the rapid quantification of benzyl chlorophenol.

The invention provides a PEDV (porcine epidemic diarrhea virus) N protein monoclonal antibody as well as a preparation method and application thereof, and the method comprises the following steps: performing prokaryotic expression on PEDV N protein, and purifying to obtain the PEDV N protein; emulsifying the purified PEDV N protein and an isopyknic Freund's adjuvant, injecting the emulsified PEDV N protein into 8-week-old female BALB / c mice at multiple points, and immunizing once every two weeks; the method comprises the following steps: uniformly mixing splenocytes of a mouse subjected to boosted immunization for 3 days with SP2 / 0 cells in a logarithmic phase according to a ratio of 5: 1, carrying out cell fusion under the action of PEG 1500, and culturing a fused cell mixed solution by using an HAT culture medium; screening out positive pores with strong antibody secretion capacity by adopting an indirect ELISA (Enzyme-Linked Immunosorbent Assay) method, and carrying out subcloning by adopting a limited dilution method to obtain six cell strains for secreting antibodies; the PEDV N protein mAb is prepared by adopting an in-vivo ascites inducing method. The prepared PEDV N protein monoclonal antibody can be specifically and efficiently combined, and can be used as an antibody tool for researching PEDV virus infection and anti-infection mechanisms.

The invention discloses a stem cell-based microvesicle as well as a preparation method and application thereof, belongs to the technical field of biological tissue engineering, and particularly relates to a preparation method of the stem cell-based microvesicle, which comprises the following steps: culturing stem cells until the cell fusion degree is 80-90%, performing hypoxia treatment, and cleaning for 2-3 times; standing and culturing the cleaned stem cells, then adding a calcium ion carrier, and carrying out ultrasonic treatment; the serum-free basic culture medium containing the calcium ion carrier is immediately absorbed and removed, the cells are cleaned through PBS, culture is conducted for 24 h, and cell supernatant is collected and merged respectively; performing first centrifugation to remove cell debris; performing second centrifugation to remove dead cells and large cell debris; carrying out third centrifugation to remove organelles; fourth centrifugation is carried out, large vesicles are removed, and a final supernatant is obtained; and filtering and centrifuging to obtain a microvesicle precipitate. The preparation method of the microvesicles aims at systematically improving the yield and biological activity of the stem cell microvesicles through low-oxygen pre-stimulation, ultrasonic-calcium ion carrier synergistic induction and differential centrifugation.