40 results about "Spleen cell" patented technology

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 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 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.

The present invention provides the preparation, epitope identification and application of a PRRSV non-structural protein 12 antibody. By constructing a eukaryotic expression vector pCDNA3.4-Nsp12-strep, transiently transfecting CHO cells, and purifying the PRRSV Nsp12 recombinant protein using the downstream strep tag, the purified Nsp12 is used as an antigen to immunize Balb / C mice. The spleen cells of the mice are isolated, and single specific B cells are separated by the FACS method. The antibody variable region sequences of the single B cells are obtained by RT-PCR. Recombinant expression plasmids of the heavy chain and light chain are constructed respectively, transfected into CHO suspension cells, expressed, and 2 complete anti-PRRSV Nsp12 monoclonal antibodies 1N14 and 2S18 are obtained by Protein G purification. The application scopes of the 1N14 and 2S18 antibodies and the specific antigen epitope recognized by the 1N14 antibody are further determined, and its sequence is 104 YEFTGNGEDW 113 。

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 provides a monoclonal antibody 3G1 capable of specifically recognizing CV-A5 virus and having neutralizing activity and application of the monoclonal antibody 3G1. The monoclonal antibody is prepared from a CV-A5 virus antigen immunized mouse and splenocytes of the mouse through a cell fusion technology, and amino acid sequences of three CDR regions of a heavy chain variable region of the monoclonal antibody sequentially comprise sequences as shown in SEQ ID NO.1-3; the amino acid sequences of the three CDR regions of the light chain variable region sequentially comprise sequences as shown in SEQ ID NO.4-6. The monoclonal antibody can be specifically combined with a CV-A5 virus, and is not combined with enteroviruses such as EV-A71, CV-A10, CV-A6, CV-A16 and the like. The monoclonal antibody targets conformational epitopes, has neutralizing activity, can specifically recognize CV-A5 viruses, is an ideal CV-A5 antigen detection antibody, and is beneficial to acceleration of the research and development process of hand-foot-mouth multivalent vaccines containing CV-A5 pathogens.

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 present invention relates to a composition for enhancing immunity, comprising, as an active ingredient, a mixture of a mixed extract of Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora, and a polysaccharide isolate thereof, or a fraction thereof. According to the present invention, the composition comprising, as an active ingredient, a mixture of a mixed extract of Angelica sinensis, Ligusticum wallichii, and Paeonia lactiflora and a polysaccharide isolate thereof, or a fraction thereof, can increase body weight, increase WBC number, restore spleen cell density, enhance spleen cell proliferation ability, and increase immune-related cytokines, especially for innate immunity. The composition can significantly increase the activity of NK cells, and can be used as an immunopotentiator or a food composition for immunopotentiation.

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 present invention provides a monoclonal cell line 2D3 that specifically secretes BP26 antibodies, a single-chain antibody, and a preparation method, belonging to the field of veterinary biotechnology. The present invention expresses and purifies the Brucella canis BP26 protein. The spleen cells of mice immunized with the protein are fused with SP2 / 0 cells in vitro to obtain the monoclonal cell line 2D3 that specifically secretes BP26 antibodies. Subsequently, RNA of the monoclonal cell 2D3 is extracted, reverse transcribed into cDNA, and the V sequence expressing the monoclonal antibody is amplified. L and V H , and V L and V H Gene ligation was performed to obtain a single-chain antibody gene. This gene was ligated to the pCold-TF vector and transformed into BL21 competent cells for induced expression, yielding a single-chain antibody against the Brucella canis BP26 protein. The single-chain antibody against the Brucella canis BP26 protein of the present invention can bind to the BP26 protein and is of great significance for the development of new brucellosis detection reagents.

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 discloses application of plant lactobacillus DPUL-F232 and capsular polysaccharide thereof in food allergy, and belongs to the technical field of microorganisms. The lactobacillus plantarum DPUL-F232 capsular polysaccharide disclosed by the invention can be used for relieving allergic symptoms and weight change of rats; the spleen index and rectum temperature change are reduced; the levels of serum antibodies IgE, IgG1 and IgG2a are reduced; mast cell protease and histamine levels are reduced; regulating the levels of cell factors IFN-gamma, IL-4, IL-10, IL-6 and IL-17A of serum and splenocytes; the heat-inactivated lactobacillus plantarum DPUL-F232 capsular polysaccharide is very high in stability, can be stored for a long time, is suitable for preparing anti-allergic foods or medicines, and has a relatively good application prospect.

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 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 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 culture medium and a method for constructing an immune organ-like model and application, belongs to the field of biological medicines, and mainly solves the problem that in-vitro human immune organ model construction and human immunology process simulation are difficult to realize. According to the method, human peripheral blood mononuclear cells (PBMC), thymus cells, tonsil cells, spleen cells or lymph node primary cells are inoculated in a culture plate with a specific structure, and an immune environment with a physiological three-dimensional (3-dimensional) structure is formed by applying a specific culture medium and conditions, so that long-term, efficient and convenient culture of the immune organ is realized; the immune tissue function of a human body is simulated. The technology involved in the invention can be applied to the fields of deep research of accelerated immunology mechanism, drug screening and evaluation, patient selection, disease detection and the like.

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 herbicide-resistant protein GAT hybridoma cell strain, an antibody produced by the hybridoma cell strain and application of the hybridoma cell strain, and the hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC No.46333.A preparation method of the hybridoma cell strain comprises the following steps: a) purifying a GAT recombinant protein obtained by prokaryotic expression; b) immunizing animals: using the GAT recombinant protein as an antigen to immunize BALB / c mice; c) cell fusion: collecting spleen cells of the immunized BALB / c mice and fusing the spleen cells with SP2 / 0 cells; d) cell strain establishment: performing subcloning by a limited dilution method, performing ELISA detection on the subcloning for 5-7 days, and screening the hybridoma cell strain stably secreting positive antibodies until the hybridoma cell strain is expanded, cultured and preserved again.The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for realizing detection of the herbicide-resistant protein GAT in a transgenic crop, thereby supporting the development, seed production, seed preservation and guarantee of the transgenic crop, and stably promoting industrialization of the transgenic crop and sustainable application of related transformants. gat The application discloses a herbicide-resistant protein GAT hybridoma cell strain, an antibody produced by the hybridoma cell strain and application of the hybridoma cell strain, and the hybridoma cell strain is preserved in the China General Microbiological Culture Collection Center with a preservation number of CGMCC No.46333.A preparation method of the hybridoma cell strain comprises the following steps: a) purifying a GAT recombinant protein obtained by prokaryotic expression; b) immunizing animals: using the GAT recombinant protein as an antigen to immunize BALB / c mice; c) cell fusion: collecting spleen cells of the immunized BALB / c mice and fusing the spleen cells with SP2 / 0 cells; d) cell strain establishment: performing subcloning by a limited dilution method, performing ELISA detection on the subcloning for 5-7 days, and screening the hybridoma cell strain stably secreting positive antibodies until the hybridoma cell strain is expanded, cultured and preserved again.The monoclonal antibody secreted by the hybridoma cell strain lays a foundation for realizing detection of the herbicide-resistant protein GAT in a transgenic crop, thereby supporting the development, seed production, seed preservation and guarantee of the transgenic crop, and stably promoting industrialization of the transgenic crop and sustainable application of related transformants. gat The application discloses a herbicide-resistant protein GAT hybridoma cell

This invention discloses a hybridoma cell line containing the herbicide-resistant protein GAT, its produced antibody, and its applications. The hybridoma cell line is deposited at the China General Microbiological Culture Collection Center (CGMCC), accession number CGMCC No. 46332. The preparation method includes: a) purifying prokaryotic expression to obtain recombinant GAT protein; b) immunizing animals: using the recombinant GAT protein as an antigen to immunize BALB / c mice; c) cell fusion: collecting spleen cells from immunized BALB / c mice and fusing them with SP2 / 0 cells; d) cell line establishment: subcloning using the limiting dilution method, performing ELISA detection on subclones after 5-7 days, until a stable hybridoma cell line secreting positive antibodies is selected for expansion, reculturing, and preservation. The monoclonal antibody secreted by the hybridoma cell line of this invention lays the foundation for the detection of the herbicide-resistant protein GAT in transgenic crops, thus supporting the development of transgenic crops. street The development, seed production, and preservation of genetically modified crops have ensured the survival of genetically modified organisms (GMOs). street The industrialization of genetically modified crops is progressing steadily, and the sustainable application of related transformants is advancing.

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 invention discloses an anti-Taq enzyme monoclonal antibody, a preparation method thereof and an application of the anti-Taq enzyme monoclonal antibody in hot start PCR (Polymerase Chain Reaction), the method comprises the following steps: S1, immunizing an animal: selecting a mouse, emulsifying the mouse with a wild recombinant Taq enzyme protein as an antigen and a Freund's adjuvant, then performing subcutaneous injection, immunizing for 2-6 times, and measuring serum titer at an interval of 1-4 weeks; s2, cell fusion: fusing spleen cells of the mouse with the highest titer with NS-1 myeloma cells, and screening hybridoma cells; s3, screening and cloning: screening positive clone secreting an anti-Taq enzyme antibody, and obtaining a stable cell strain through 3-4 times of subcloning; and S4, antibody purification: purifying the IgG antibody in the ascites or cell supernatant to obtain the anti-Taq enzyme monoclonal antibody. The anti-Taq enzyme monoclonal antibody provided by the invention has the characteristics of high specificity, high sensitivity, good stability, simplicity and convenience in operation and the like, and has a wide application prospect.

The invention discloses a hybrid snakehead spleen cell line as well as an establishment method and application thereof, and belongs to the technical field of fish cell culture. The hybrid snakehead spleen cell line provided by the invention is named as CAMSp and is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation number is CCTCC NO: C202598. According to the application, the spleen tissue from the hybrid snakehead is subjected to cell culture, the culture conditions of the cells are preliminarily determined, the sensitivity of the cells to the hybrid snakehead rhabdovirus (HSHRV) is detected, and a material basis is provided for researching the pathogenesis of the HSHRV, a detection method, vaccine preparation, interaction with host cells, disease prevention and control and the like.

The present invention relates to the expression of a CHO cell against the ASFVD 117L encoded protein, the preparation of monoclonal antibodies, epitope identification and application. By constructing a eukaryotic expression vector pCDNA3.4-D117L-strep, transiently transfecting CHO cells to express recombinant p17 protein, immunizing Balb / c mice after purification, fusing the spleen cells of mice with myeloma SP2 / 0 cells to prepare hybridoma cells after four immunizations, and then using the indirect ELISA method for subcloning to obtain monoclonal antibodies against p17 protein, and identifying the antigen epitope corresponding to the antibody as <supgt;8< / supgt;LLSHNLSTREGIK<supgt;20< / supgt>.

The present invention relates to a composition for enhancing immunity, which contains a mixture of a mixed extract of angelica, cnidium, and paeonia and a polysaccharide isolated therefrom; or a fraction thereof as an effective ingredient. The composition containing a mixture of a mixed extract of angelica, cnidium, and paeonia and a polysaccharide isolated therefrom; or a fraction thereof as an effective ingredient according to the present invention increases body weight, increases WBC count, restores spleen cell density, increases spleen cell proliferation ability, and increases immune-related cytokines as well as significantly increases NK cell activity, particularly with respect to innate immunity, and can be thus applied as an immune-enhancing agent or a food composition for enhancing immunity.