149 results about "BALB/c" patented technology

The invention discloses preparation of a human CD56 immune phage display antibody library, which comprises the following steps: S1, animal immunization: expressing recombinant hCD56 protein in escherichia coli, then immunizing a Balb / c mouse, and detecting immune titer; s2, splenocyte preparation: extracting RNA (Ribonucleic Acid) of immune mouse spleen cells, and performing reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid); s3, preparation of an initial bacteriophage display library: amplifying a heavy chain variable region and a light chain variable region by using a PCR technology, and then assembling to form scFv; constructing scFv phagocytids, electrically transforming the scFv phagocytids into TG1 competent cells, and constructing a bacterial library; s4, constructing a phage single-chain library of the hCD56: infecting the bacterial library by using a helper phage M13K07, and constructing a phage single-chain antibody library; and S5, elutriation and screening of a phage library: specifically enriching a phage single-chain antibody library, screening positive clones from the phage single-chain antibody library, and determining a sequence. According to the invention, the human hCD56 antibody scFv sequence fragment can be screened out, and the research and development period is shortened.

The invention discloses a monoclonal antibody targeting all CD45 subtypes and application of the monoclonal antibody, and belongs to the field of monoclonal antibody preparation. The hybridoma cell strain capable of stably secreting all anti-human CD45 subtype antibodies is successfully obtained by stably expressing short and long molecular subtypes of human CD45 extracellular regions in L929 mouse fibroblasts, immunizing BALB / c mice with L929 cells expressing the two molecular subtypes of human CD45, and fusing splenocytes with SP20 myeloma cells after the serum titer reaches the standard. And the monoclonal antibody targeting all subtypes of CD45 is obtained. The monoclonal antibody provided by the invention shows a high-affinity binding characteristic with all human CD45 molecular subtypes, has important application value in the aspect of human CD45 molecular detection, and can be used as a therapeutic antibody in the fields of tumor immunotherapy, autoimmune disease treatment, transplant rejection resistance and the like.

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 relates to the technical field of antibodies, in particular to an anti-African swine fever virus p30 protein monoclonal antibody and application thereof, and the anti-African swine fever virus p30 protein monoclonal antibody can be specifically combined with African swine fever virus p30 protein. The anti-ASFV p30 protein monoclonal antibody is prepared by immunizing a BALB / c mouse through an immunological method, and the antibody is high in specificity, titer and affinity and can be applied to immunological detection or antibody detection kits.

The monoclonal antibody of the glycoprotein Gn of the severe fever with thrombocytopenia syndrome virus is screened on the basis of immune repertoire sequencing, and the biological characteristics of the monoclonal antibody are identified. The method comprises the following steps: taking purified SFTSV-Gn as bait, taking virus specific B cells from an immunized female BALB / c mouse B cell bank, selecting an antibody sequence with potential binding activity by utilizing a 10 * immune repertoire sequencing method in combination with bioinformatics analysis, constructing the antibody sequence to an eukaryotic expression vector pcDNA3.4, expressing and purifying a monoclonal antibody, and identifying the biological activity of the monoclonal antibody. 14 antibodies are selected from 3544 captured B cells, functional verification shows that 12 antibodies can be combined with SFTSV-Gn glycoprotein, and the Gn-5 antibody and the Gn-7 antibody have high combining capacity; western blot, indirect immunofluorescence test and flow cytometry prove that the antibody can specifically recognize the SFTSV, the effective concentration is as low as 2 mu g / mL, and a foundation is laid for development of a rapid diagnosis reagent for the SFTSV.

The invention belongs to the technical field of biology, and particularly relates to a human metapneumovirus mouse adapted strain and application thereof. Compared with a wild strain, the strain has higher susceptibility to animals; the strain can be used for treating BALB / c mice and can cause typical clinical symptoms and even death, for example, after the strain infects the BALB / c mice, the disease symptoms are obvious, the weight of 83% infected mice is severely reduced to 75% or below and reaches the humanity killing standard, and the death rate reaches 50% or above 4 days after the strain infects the BALB / c mice; the strain can be used for constructing a cell model or an animal model of human metapneumovirus infection or diseases caused by the human metapneumovirus infection, and is further used for screening drugs for preventing and / or treating the human metapneumovirus infection or the diseases caused by the human metapneumovirus infection. In addition, the strain can also be used for preparing medicines for preventing and / or treating human metapneumovirus infection or diseases caused by the human metapneumovirus infection, preparing antibodies or antiserum for resisting the human metapneumovirus and the like.

The invention belongs to the technical field of biological medicines, and particularly relates to application of ganoderic acid A in preparation of a medicine for preventing and / or treating biliary atresia. The new application of ganoderic acid A in preparation of drugs for preventing and / or treating biliary atresia is found for the first time. The method comprises the following steps: within 24 hours after a BALB / c newborn mouse is born, inducing the formation of BA by intraperitoneal injection of RRV, and from the second day after intraperitoneal injection of RRV, carrying out intraperitoneal injection of Ganoderic Acid A on the mouse every day until the twelfth day, it can be observed that the survival time of BA mice treated by the Ganoderma Acid A is prolonged, the weight loss is obviously improved, the jaundice symptom is relieved, the liver function index is obviously improved, the infiltration number of inflammatory cells around the intrahepatic bile duct is obviously reduced, and the liver bile duct injury is relieved. The invention provides a new potential target and a treatment strategy for treatment of BA, and has potential clinical treatment significance.

The invention discloses application of piperlongumine in preparation of a medicine for treating virus-induced myocardial fibrosis. According to the application, an in-vivo myocardial fibrosis model is constructed by infecting a BALB / c mouse with a CVB3 virus, and meanwhile, an in-vitro cell model for proliferation and differentiation of primary myocardial fibroblasts is constructed by inducing TGF-beta1. Results prove that when the piperlongumine is applied to the acute stage (1-14 days), the chronic stage (14-28 days) and the whole course (1-28 days) of the viral myocarditis, the myocardial fibrosis of the viral myocarditis can be remarkably improved, the cardiac function is improved, and the curative effect of the piperlongumine applied to the whole course is superior to that of the piperlongumine applied to the acute stage and the chronic stage; the specific mechanism is that Smad2 / 3 and p-38 / YAP pathways jointly play a role in inhibiting myocardial fibrosis.

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 present application provides a mouse model for studying microsatellite stable and unstable intestinal cancer. Specifically, the present application provides the use of a hybrid mouse in a reagent or kit for preparing a mouse model animal for constructing MSS type intestinal cancer and MSI-H type intestinal cancer, wherein the mouse is a F1 generation of C57BL / 6 x BALB / c. The mouse model of the present application provides the same genetic background for MSS type intestinal cancer cells and MSI-H type intestinal cancer cells, making the research of MSS type intestinal cancer and MSI-H type intestinal cancer more scientific.

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 belongs to the technical field of biology, and particularly relates to a multi-respiratory-tract pathogen mixed infection animal model and an establishment method and application thereof. The construction method provided by the invention comprises the following steps: firstly, infecting a female BALB / c mouse by using an influenza virus and mycoplasma pneumoniae virus mixture to construct a mycoplasma viral infection animal model, infecting for 20-28 hours, and finally successfully constructing a mixed infection mouse model. The state of the infected mouse is further observed, and the weight and death conditions, the clinical symptoms and tissue pathological changes of the infected animal model and the like are recorded. Therefore, the construction method provided by the invention is simple, and the constructed animal model is a safe, effective and economical tool and can be used for rapid preclinical evaluation of the screening process of medical products such as respiratory tract mixed infection detection kits, vaccines, medicines and the like under biosafety level 2 laboratory conditions.

The application belongs to the technical field of biotechnology, and particularly relates to a human metapneumovirus mouse-adapted strain and application thereof. Compared with wild strains, the strain can exhibit stronger susceptibility to animals, and can cause typical clinical symptoms and even death. For example, after the strain infects BALB / c mice, the disease symptoms are obvious, 83% of the mice have a severe decrease in body weight to below 75% after infection, and have reached the humane killing standard. Four days after the strain infects BALB / c mice, the mortality rate is above 50%. The strain can be used to construct a cell model or an animal model of human metapneumovirus infection or a disease caused by the human metapneumovirus, and is further used to screen drugs for preventing and / or treating human metapneumovirus infection or a disease caused by the human metapneumovirus. In addition, the strain can also be used to prepare drugs for preventing and / or treating human metapneumovirus infection or a disease caused by the human metapneumovirus, and to prepare antibodies or antisera against the human metapneumovirus.

This invention provides a peanut allergen peptide, its screening method, and its application. Phage display technology is used to screen for conformational epitopes of Ara h5 and Ara h8 proteins. Polyclonal IgE and IgG antibodies against the target proteins are prepared by immunizing BALB / c mice with Ara h5 and Ara h8 proteins as antigens. ELISA and dotted immunoblotting detection revealed that the titers of polyclonal IgG and IgE against Ara h5 and Ara h8 proteins were 1:64000 and 1:320, respectively, and the antibodies exhibited specificity. Using the antibodies as targets, phage display technology screened the conformational epitope sequences of Ara h5 protein as WETIYSR and FHWWYLK. The conformational epitopes of Ara h8 protein were FPYMKFV, FPYMKFR, SMFARID, and SFHWWLF.

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.

This invention belongs to the field of animal model construction technology, specifically relating to a mouse immunosuppressive model that maintains a long-term immunosuppressive state, its construction method, and its application. This invention establishes a stable immunosuppressive state model with reduced neutrophil count and no significant decrease in erythrocyte count by administering different doses of immunosuppressants (10-50 mg / kg body weight) to Balb / c mice via a combination of "reduced total dose + multiple gavage administration + satellite monitoring." The method for constructing this mouse immunosuppressive model reduces drug usage, controls neutrophil count rebound, establishes a stable model of reduced neutrophil count, and avoids deep bone marrow suppression (i.e., does not cause an uncontrollable decrease in erythrocyte or platelet count), which is beneficial for long-term experimental conduct and drug evaluation.

A recombinant humanized collagen type III alpha-1 (rhCol III a1), and an expression vector and use thereof are provided, relating to the technical field of bioengineering. A sequence on positions 154 to 1,221 of a human type III collagen α chain is ligated to an optimized vector pcDNA3.1, a resulting optimized expression vector is transfected into a human embryonic kidney cell Expi293F, and then rhCol III a1 is secreted to form a fusion protein with a soluble green fluorescent protein (GFP). The expressed rhCol III a1 that forms the fusion protein with the GFP shows an activity of promoting the migration of BALB / c 3T3 cells, thus exhibiting a great application potential in the field of skin care.

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 present application relates to the field of biotechnology and in vitro diagnosis, and relates to a monoclonal antibody of eosinophil peroxidase and application thereof. The antibody of the present application is mainly used for detecting the content of eosinophil peroxidase by western blot. The monoclonal antibody of eosinophil peroxidase of the present application is prepared by immunizing Balb / c mice with recombinant eosinophil peroxidase as an immunogen, then fusing the spleen with myeloma cells NS1, and screening and purifying. The prepared monoclonal antibody of eosinophil peroxidase specifically recognizes the linear epitope of eosinophil peroxidase.

The present application relates to in situ model modeling method, especially for a kind of non-invasive cervical cancer in situ model modeling method of mouse, including selecting several 8 weeks old BALB / c-nude mouse, all mice are divided into non-invasive group and operation group, the age, weight of two groups of mice are compared, and there is no statistically significant difference;Non-invasive group is widened with the cotton stick of diameter 3mm and wipes dry vagina, the tumor block with luciferase fluorescence is immersed in modified hydrogel, after heating and making modified hydrogel solidification, the tissue glue mixed with hard gypsum in modified hydrogel is dropped on the surface of modified hydrogel and quickly inserted into vagina, when there is obstruction, stop deepening, pause 30s and wait for tissue glue to be completely dry, when tumor block is fixed in cervix, it is modeling completion;Operation group first opens abdominal cavity, and tumor tissue of same size is sutured on cervix.This kind of non-invasive cervical cancer in situ model modeling method of mouse, using non-invasive method to replace operation method to establish non-invasive cervical cancer in situ model, can reduce the pain of mouse, reduce experimental difficulty.

The invention discloses a plant lactobacillus AR195 strain and application thereof in preparation of a medicine for preventing and / or treating lung cancer, and belongs to the technical field of microorganisms. Through verification, the plant lactobacillus AR195 can inhibit the growth of lung cancer cells, also has certain prevention and treatment effects in a lung cancer mouse model, and can reduce the tumor burden of a BALB / c mouse lung cancer model by reducing MMP9 secretion, so that the plant lactobacillus AR195 can be used for preventing and assisting in treating lung cancer. Meanwhile, the accumulation of the AR195 bacteria in the lung can be improved to the maximum extent through atomization administration, and meanwhile, the adverse side effects of the whole body are reduced. A new drug source is provided for treatment of lung cancer, a novel treatment strategy is provided for treatment of lung cancer by probiotics, and treatment of lung cancer and development of a novel treatment method are facilitated. Meanwhile, the application range of the plant lactobacillus AR195 strain is widened, and a novel treatment strategy is provided for a potential mechanism of treating lung cancer by probiotics.

The present application belongs to the technical field of bioengineering, and particularly relates to a Claudin18.2 gene engineering antibody based on DNA immunization and a preparation method and application thereof. The Claudin18.2 gene engineering antibody comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises any one of the amino acid sequences shown in SEQ ID NO. 15-16; and the light chain variable region comprises the amino acid sequences shown in SEQ ID NO. 19-20. The Claudin18.2 gene engineering antibody is obtained by immunizing Balb / c mice with full-length plasmid DNA and full-length plasmid DNA+extracellular segment polypeptide as immunogens respectively, obtaining plasma cells capable of secreting specific antibodies against Claudin18.2, and then preparing specific mouse-derived gene engineering antibodies through single-cell optical guidance technology and genetic engineering technology. The gene engineering antibodies have high titer and strong specificity, and have specific recognition ability for natural protein Claudin18.2.

The invention discloses a polyclonal antibody of panda Rbp7 protein as well as a preparation method and application thereof, and belongs to the technical field of antibodies. The first to nineteenth amino acid sequences of the panda Rbp7 protein are used as antigen polypeptides, and are specifically shown as SEQ ID No.1. After chemical synthesis or exogenous expression purification, a Balb / C mouse is immunized, and the polyclonal antibody is obtained through repeated immunization, serum titer detection and affinity chromatography purification. The titer of the antibody is not lower than 1 / 128000, the panda Rbp7 recombinant protein can be efficiently and specifically detected through WB and ELISA, the blank of a protein detection tool is filled, and reliable technical support is provided for research on the functional mechanism of the panda Rbp7 protein.

To conduct detection of Salmonella, serotype transversely and specifically relative to other strains.SOLUTION: A heat-killed bacterial cell of Salmonella and antigen liquid containing an adjuvant are inoculated in a BALB / c mouse. After the inoculation, splenocytes or lymph node cells of the mouse and P3U1 are fused to prepare a hybridoma, and therefrom, a hybridoma for producing a monoclonal IgG antibody showing Salmonella specificity is selected, whereby three hybridomas for producing superior Salmonella specific antibodies are successfully acquired. Next, these monoclonal antibodies are used to detect Salmonella of a plurality serotypes and other strains according to sandwich ELISA, thereby obtaining a result in which Salmonella is detected serotype transversely, whereas other strains are not detected.SELECTED DRAWING: Figure 1

The invention provides a method for reducing PRV virulence through gene insertion and application of the method in vaccine preparation, and belongs to the technical field of gene engineering. The invention provides a method for reducing the virulence of a pseudorabies virus. An EGFP (Enhanced Green Fluorescent Protein) protein expression cassette is inserted into the upstream of a UL40 gene. According to the invention, after the PRV is subjected to the genetic engineering modification, the pseudorabies virus attenuated strain EGFP-PRV can be obtained through virus rescue. Compared with a wild type PRV-DX, the pathogenicity is weakened, no pathological clinical symptom appears after a Balb / c mouse is infected with the pseudorabies virus, and the pseudorabies vaccine has the prospect of preparing the pseudorabies vaccine.

Provided in the present disclosure are a compound for inhibiting the expression of the LPA gene, a pharmaceutical composition and the use thereof, which belong to the technical field of small nucleic acid drug delivery. The oligonucleotide-conjugated compound and the pharmaceutical composition thereof provided in the present disclosure can significantly inhibit the expression of LPA mRNA in animal liver tissues. The oligonucleotide-conjugated compound and the pharmaceutical composition thereof provided in the present disclosure are helpful for relieving, preventing and / or treating diseases or conditions mediated by LPA gene expression dysregulation.AA Relative expression level of LPA mRNA (%)BB Level of LPA mRNA in liver tissues of Balb / c-HDI miceCC s.c. single dose, 1 mg / kg

The invention provides a recombinant respiratory syncytial virus vaccine as well as a preparation method and application thereof, and belongs to the technical field of viral vaccine preparations. Wherein the recombinant respiratory syncytial virus vaccine is a vaccine preparation in a liquid dosage form comprising pre-F protein antigen; wherein the amino acid sequence of the pre-F protein antigen is any one of the amino acid sequences as shown in SEQ ID NO. 1 to SEQ ID NO. 3. The recombinant respiratory syncytial virus vaccine provided by the invention is a liquid vaccine preparation comprising a recombinant respiratory syncytial virus (RSV) protein stock solution, the vaccine preparation has better stability, and the self-made recombinant respiratory syncytial virus vaccine is proved to have good immunogenicity through immune experiment research on a BALB / c mouse by intramuscular injection.

The present invention belongs to the technical field of vaccines and relates to an encephalomyocarditis multi-epitope recombinant protein, and the encephalomyocarditis multi-epitope recombinant protein comprises 8 B-cell epitopes and 1 signal peptide. The present invention also provides the application of the recombinant protein in the preparation of an encephalomyocarditis vaccine. With the goal of developing a safe and effective vaccine, the B-cell epitopes of VP1, VP2, and VP3 of the EMCV structural protein are screened, and a recombinant epitope protein with good hydrophilicity, strong antigenicity, and high surface accessibility is constructed. By immunizing BALB / c mice, it is found that it can induce the production of neutralizing antibodies and a Th1-dominated cellular immune response, and can protect the mice from EMCV attack. The results show that the MIgH-EMCV-II<supgt;2D2< / supgt; protein may be a potential candidate for developing a safe and effective multi-epitope vaccine to control EMCV infection.

This application relates to a traditional Chinese medicine composition for treating eczema, its preparation method and uses. The traditional Chinese medicine composition of the present invention is derived from inheriting the classic ancient formula "Xiaofeng Powder" and combining with the traditional Chinese medicine theory of "tonifying the earth", which conforms to the traditional Chinese medicine theory, and is mainly used for treating eczema, especially acute eczema. The traditional Chinese medicine composition of the present invention can play an obvious anti-eczema effect on the eczema model of Balb / c mice, and the effect increases with the increase of the dose. The curative effect on erythema, epidermal exfoliation and lichenification of the eczema model animals is significantly better than that of the commonly used proprietary Chinese medicine Xiaofeng Zhiyang Granule for this indication.