19 results about "Spleen cell" patented technology

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

The invention discloses a chikungunya virus envelope E2 protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

The invention discloses a yellow fever virus envelope protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. Yellow fever virus envelope protein E expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the yellow fever virus envelope protein E are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the monoclonal antibody with high neutralizing activity and capable of completely protecting mice from yellow fever virus lethal attack is obtained by using the monoclonal antibody as a template, connecting the monoclonal antibody and the constant region gene to an expression vector, carrying out mammalian cell expression and purification, and carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse virus attack experiment, such that the monoclonal antibody has high neutralizing activity and can completely protect mice from yellow fever virus lethal attack; the monoclonal antibody has application value in prevention and treatment of yellow fever.

The invention discloses a chikungunya virus monoclonal antibody and application thereof in preparation of preventive and therapeutic antibody drugs, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

The application discloses application of pulegone as an effective component in an immunosuppressant, and the pulegone with significant calcineurin (CN) inhibiting activity is screened from the Schleichera oleosa Kurz, the immunosuppressive effect of the pulegone is equivalent to that of cyclosporine A which is widely used in clinic, mechanism research shows that the pulegone targets CN / NFAT signal path to play the immunosuppressive activity, and the pulegone has very weak cytotoxicity to spleen cells, and is a high-efficiency and low-toxicity immunosuppressant.

The invention relates to the fields of therapy and drug delivery. It is based on an unexpected finding that a combination of an oligonucleotide-based medicament with a saponin component comprising a penta-cyclic triterpene saponin of a 12,13-dehydrooleanane aglycone core, not only does not appear to be associated with oligonucleotide medicament-induced nephrotoxic effects after systemic administration in vivo, but also that it allows the oligonucleotide-based medicament to enter and act on its nucleic acid target inside of the kidney cells instead of remaining unproductively trapped in renal subcellular compartments or being eliminated into the urine via the renal glomerular filtration system. In line with this finding, herein disclosed are therapeutic combinations, compositions, and formulations, as well as methods of treatment involving their administration, which comprise an oligonucleotide-based medicament that acts on an intracellular nucleic acid target in the kidney cells and / or in the cells of the spleen, being the other blood filtering organ in addition to the kidney, and a saponin component that enables the effective release of said medicament inside of these cells, thus allowing its use at a much lower and safer dose. The provision of the presented herein therapeutic combinations enables effective modulation of gene expression in the kidney and / or spleen cells and, therefore, it opens not only new treatment options for kidney diseases, in particular those involving inflammation and / or splenomegaly, but also for diseases of other organs which affect or are affected by the pathological processes happening in the kidneys and / or in the spleen, in particular being inflammatory processes.

The application provides a hybridoma cell strain secreting a monoclonal antibody against a alginate mannuronic acid tetrasaccharide epitope, a monoclonal antibody and application. In order to enhance the immunogenicity of the mannuronic acid tetrasaccharide, a glycoconjugate KLH-1 is used as an immunogen to inject and immunize a mouse, so that the mouse is stimulated to produce a specific immune response against the mannuronic acid tetrasaccharide epitope, then spleen cells of the immunized mouse are fused with myeloma cells, and after screening, an initial hybridoma cell strain is obtained; then after subcloning and specific screening, a hybridoma cell strain capable of stably secreting a monoclonal antibody of the target mannuronic acid tetrasaccharide epitope in alginate is obtained, and a monoclonal ascites antibody is further obtained. The monoclonal ascites antibody shows specific recognition and combination ability for pseudomonas aeruginosa, and the combination activity is related to the expression level of alginate on the surface of the bacteria. Therefore, the monoclonal ascites antibody can be used as a precise detection and diagnosis tool for pseudomonas aeruginosa, and can also be applied to the antibacterial treatment of pseudomonas aeruginosa infection.

The invention discloses a method for preparing a high-titer polyclonal antibody from a Zika virus strain, which comprises the following steps: selecting the Zika virus strain, culturing in cells, and purifying virus particles with high purity to prepare an immunogen; an optimized animal immunization strategy is adopted and comprises the steps of selecting a proper host animal, applying a potent immunologic adjuvant, namely a Freund's adjuvant, and designing a scientific immunization program, so that humoral immune response of the host is stimulated to the maximum extent; when the titer of the antibody reaches a peak value, collecting antiserum, synthesizing hybridoma cells through splenocytes, and obtaining the high-purity polyclonal antibody through a multi-step purification technology. According to the method, high-purity and non-chemically-inactivated whole virus particles which retain natural conformation are used as immunogens, a mode of primary immunization and multiple booster immunization is adopted, a Freund's adjuvant is combined, animals are continuously stimulated to generate high-titer antibodies, non-specific antibodies and impurities are effectively removed through multi-step purification, and the immunogen immunogen is obtained. The target antibody with high purity and high specificity is obtained.

The invention specifically discloses an application of bacillus amyloliquefaciens E35 in preparation of a product for resisting grouper iridovirus infection, and the application of the bacillus amyloliquefaciens E35 in preparation of a product for resisting grouper iridovirus infection. The bacillus amyloliquefaciens E35 is preserved in Guangdong Microbial Culture Collection Center on December 20, 2024, and the preservation number is GDMCC 65657. According to the application of the bacillus amyloliquefaciens E35 in preparing the product for resisting grouper iridovirus infection, the effect of preventing and treating the grouper iridovirus is excellent, the infection activity of the grouper iridovirus on spleen cells is reduced, the death rate of groupers infected with the iridovirus is reduced, the disease time is delayed, and the safety of the groupers is improved. The drug resistance of fishes can be reduced, and toxic and side effects on fish bodies due to prevention and treatment can be reduced.

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 lipid nanoparticle (LNP) compositions, and diagnostic or therapeutic polynucleotides, e.g. TERT mRNA, which may be delivered with the LNP compositions in formulations to various tissues and cell types throughout the mammalian body, including stem cells, progenitor cells, germ cells, differentiated cells, or terminally differentiated cells, cancer cells, endothelial cells, epithelial cells, spleen cells, hepatocytes, kidney cells and / or bone cells, e.g., for the diagnosis, prevention and / or treatment of conditions or disease.

The invention discloses a chikungunya virus envelope E2 protein monoclonal antibody and application of the chikungunya virus envelope E2 protein monoclonal antibody in preparation of preventive and therapeutic antibody drugs, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

The invention discloses an anti-p53R175H / HLA-A 02: 01 monoclonal antibody, a hybridoma cell strain S2-C33 for preparing the monoclonal antibody as well as a preparation method and application of the monoclonal antibody. The preparation method of the hybridoma cell strain comprises the following steps: immunizing a BALB / c mouse by taking human p53R175H / HLA-A 02: 01 recombinant protein as an immunogen to prepare an immunized mouse; taking splenocytes of the immunized mouse, and fusing the splenocytes with the SP2 / 0 myeloma cells to prepare fusion cells; screening monoclonal antibody positive fusion cells capable of secreting a compound specifically combined with p53R175H / HLA-A 02: 01 from the fusion cells, wherein the monoclonal antibody positive fusion cells are the hybridoma cell strain. The monoclonal antibody secreted by the hybridoma cell strain can be specifically combined with p53R175H / HLA-A 02: 01 presented on the surface of cancer cells, and the specificity is relatively high.

The application discloses a Plectropomus leopardus spleen cell line and application thereof. Plectropomus leopardus The spleen cell line PLS has a preservation number of GDMCC No: 68131. The PLS is obtained through primary culture and multiple subculture, has a fast cell growth speed, has been stably subcultured for more than 60 generations, has good activity after cryopreservation, and lays a foundation for Plectropomus leopardus gene germplasm resource preservation. Meanwhile, the PLS is sensitive to fish nervous necrosis virus and iridovirus, can be applied to exogenous gene function research, fish virus isolation, virus infection pathogenesis, virus host interaction and virus in-vitro infection model, and provides a high-efficiency amplification system for Plectropomus leopardus virus vaccine preparation.

The application provides a semi-labdanum type diterpene alkaloid compound and a preparation method and application thereof. The application provides three semi-labdanum type diterpene alkaloid compounds with new skeletons, wherein the semi-labdanum type diterpene alkaloid compounds are three new skeleton compounds Diaporlabanoids A-C with immunosuppressive activity which are separated from an endophytic Diaporthe phaseolorum H3-2 fungus in a mangrove plant Xanthophyllum coriaceum. The application also carries out experimental research on cytotoxicity and immunosuppressive activity of the separated compounds, and the results show that Diaporlabanoid A has strong immunosuppressive activity on ConA-induced mouse T lymphocytes and low cytotoxicity on normal spleen cells, and can be used for preparing immunosuppressive drugs targeting two cascade signal pathways of CaN / NFAT and PKC theta / NF-kappa B. The semi-labdanum type diterpene alkaloid compound provided by the application can be applied to preparation of immunosuppressive drugs.

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

The invention discloses a yellow fever virus monoclonal antibody and application thereof, and belongs to the technical field of medicines. Yellow fever virus envelope protein E expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the yellow fever virus envelope protein E are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the monoclonal antibody with high neutralizing activity and capable of completely protecting mice from yellow fever virus lethal attack is obtained by using the monoclonal antibody as a template, connecting the monoclonal antibody and the constant region gene to an expression vector, carrying out mammalian cell expression and purification, and carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse virus attack experiment, such that the monoclonal antibody has high neutralizing activity and can completely protect mice from yellow fever virus lethal attack; the monoclonal antibody has application value in prevention and treatment of yellow fever.

The invention discloses variable region amino acid sequences of two monoclonal antibodies of yellow fever virus non-structural protein NS1 and application of the variable region amino acid sequences, and belongs to the technical field of medicines. Yellow fever virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the yellow fever virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. The method comprises the following steps of: constructing a monoclonal antibody, connecting the monoclonal antibody and a constant region gene into an expression vector, expressing and purifying by mammalian cells, and detecting by enzyme-linked immunosorbent assay to obtain two monoclonal antibodies with high affinity and independent epitopes, and the two monoclonal antibodies are not combined with non-structural proteins NS1 of other eight viruses of flaviviridae flaviviruses. The two monoclonal antibodies have application value in diagnosis and prevention of yellow fever.