259 results about "Mouse Spleen" patented technology

The invention discloses a human SARS-CoV-2 monoclonal antibody. The preparation method of the human SARS-CoV-2 monoclonal antibody comprises the steps: adopting SARS-CoV Nucleocapsid recombinant protein as immunogen, immunizing BALB/c mice, performing fusion and subcloning on spleen cells and myeloma cells of mice, then performing a large amount of repeated screening and domestication of cell lines through commercialized products SARS-CoV-2 Nucleocapsid and MERS Nucleocapsid so as to obtain a hybridoma cell line capable of secreting the SARS-CoV-2-resistant N monoclonal antibody with high affinity and high specificity finally and successfully, and finally performing ascites preparation and purification so as to obtain the monoclonal antibody, wherein the amino acid sequence of the SARS-CoVNucleocapsid recombinant protein is shown in SEQ ID No. 1. The invention also discloses application of the monoclonal antibody in preparation of SARS-CoV-2 virus detection products and preparation ofdrugs for inhibiting the SARS-CoV-2 viruses. The monoclonal antibody can be used for detecting the SARS-CoV-2 in human throat swabs/pulmonary secretions and other samples by using a double-antibody sandwich method, and can be applied to diagnosis and prevention and control of SARS-CoV-2 virus infection and scientific researches of viruses and other study.

The invention belongs to the technical field of genetic engineering, and in particular relates to screening of a single-chain antibody against fumonisin and application thereof in immunoassay of fumonisin. The screening method comprises the following steps of: directly setting off from coupled antigen FB1-KLH(keyhole limpet hemocyanin) immunized mouse spleen cells, establishing a single-chain antibody gene library by a molecular cloning method and technology, then screening by a phage display technology, expressing ELISA(enzyme-linked immuno sorbent assay) test, sequencing, and finally obtaining the single-chain antibody against fumonisin named as FB-Mu 1H3 with high affinity and a coding gene thereof. The single-chain antibody can be directly applied to assay of fumonisin after being expressed and purified in a large quantity in escherichia coli, and the applications include assay of fumonisin pollution in field crops, feed, grain or food.

The invention belongs to the technical field of biomedicines, and in particular relates to application of seaweed polysaccharides. The seaweed polysaccharides can be taken as preparations for preparing anti-viral or immune medicines. Cell models and in-vivo animal experiments show that the seaweed polysaccharides can be used for significantly enhancing the animal immunity, and can also be used for promoting the generation of cell factors, the typing of T lymphocytes and the proliferation of mouse spleen cells so as to ensure that cellular immune reaction can be activated. The functional seaweed polysaccharides disclosed by the invention are natural polysaccharides which are extracted from seaweeds such as large seaweeds and kelps, gulfweeds, grateloupia sparsa, eucheuma muricatum, ulva pertusa kjellm, enteromorpha, gracilaria, ascophyllum nodosum and scytosiphon lomentaria, and can also be low-molecular-weight seaweed polysaccharides or seaweed oligosaccharides which are prepared by degrading the seaweed polysaccharides by virtue of different preparation methods. According to the application disclosed by the invention, a polysaccharide extract of single seaweed or a polysaccharide extract mixture of a variety of seaweeds can be taken as a novel anti-viral and immune enhancing agent to be applied to feeds of livestock, poultry, fish, shrimps and shellfish, and has wide application prospects.

A medicine combination has the function of resisting lung cancer. The combination is mixed with rhizoma paridis saponin and milk vetch root amylose according to the weight ratio of 3 to 1. The rhizoma paridis is extracted by alcohol and then the rhizoma paridis extract is obtained through gradient elutriation of macroporous absorption resin alcohol; the milk vetch root is extracted by water and then the protein is settled and removed to get the milk vetch root amylose; the two extracts are mixed and the medicine is made. The MTT activities experiment in vitro with the MTT method has proven that the medicine combination can obviously control the growth of various lung cancer cells such as LA795 lung adenocarcinoma cell and the IC₅₀ can reach 26.73 ug/ml; the experiment of mouse with the lung cancer tumor has represented that the tumor constraint rate can reach 55.63 percent; in this way, lung transfer of hypodermic transplanted tumor of the mouse with the lung cancer tumor can be obviously controlled and the tumor cells can be brought to death; moreover, spleen index and thymus index can be promoted and the medicine is innocuous and has no side effect. In addition, compared with the raw materials, the medicine combination has high activity and clear function; moreover, the medicine can be made into different types.

The invention discloses a GPC3 (glypican-3) monoclonal antibody hybridoma cell strain 7D11 and a preparation method and application thereof, which belong to the technical field of bioengineering. The GPC3 monoclonal antibody hybridoma cell strain 7D11 has a preservation code of CGMCC No.5426. The preparation method of the hybridoma cell strain 7D11 includes: using GPC3 protein as an immunogen to immunize a mouse, subjecting spleen cells of the mouse with serum titer more than 1:104 and myeloma cells SP2/0 to fusion, using a HATRPMI-1640 medium to screen fused cells, screening by ELISA (enzyme-linked immuno sorbent assay) and multiple limiting dilution process, and finally obtaining the hybridoma cell strain 7D11. The GPC3 monoclonal antibody hybridoma cell strain 7D11 is high in yield of secreted antibodies and easy to survive, the secreted monoclonal antibodies are high in titer, flexible in reaction, easy to detect, low in production cost and widely applicable to detection of GPC3 protein expression.

The invention discloses a preparation method and application of a product obtained by coupling melamine with carrier protein, as well as a melamine antibody. The product obtained by coupling the carrier protein with the melamine is used as artificial antigen and applied to an immunological method for melamine detection. The preparation method comprises the following steps of immunizing animals with the coupled product so as to prepare an antibody used for melamine detection, fusing BALB/C mouse spleen cells immunized with the coupled product and SP2/0 mouse myeloma cells, obtaining hybridoma capable of stably transferring culture and secreting anti-melamine specific monoclonal antibodies by screening positive hybridoma and cloning cells, and preparing an ascites monoclonal antibody. The prepared monoclonal antibody is utilized to establish a direct competitive ELISA method having high specificity, sensitivity and accuracy to the melamine, as well as an immune colloidal gold test strip. The preparation method for the product obtained by coupling melamine with carrier protein, as well as the melamine antibody provides service for the rapid detection of melamine-type residue in foods.

The invention relates to a hybridoma cell strain secreting thiamethoxacin monoclonal antibody and application thereof, belonging to the field of food safety immunodetection. The accession number of the hybridoma cell strain is CGMCC No. 14699. According to the invention, a complete Freund's adjuvant is used for primary immunization of a BALB/c mouse, then an incomplete Freund's adjuvant is used for booster immunization three times, and a thiamethoxam complete antigen containing no adjuvant is used for impact immunization once, so the BALB/c mouse is immunized; and then the high-titer low-IC50spleen cells of the immunized mouse are fused with mouse myeloma cells by using a PEG method, and then the cell strain is obtained through indirect competitive ELISA screening and subcloning three times. The monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (with an IC50 value of 0.81 ng/mL) to thiamethoxam and can be used for detection of thiamethoxamresidues in food.

The invention relates to an imidacloprid pesticide against monoclonal antibody and a preparation method thereof, and belongs to the technical field of biology. The preparation method comprises the following steps of: immunizing a BALB/c mouse by using a coupling substance of immune hapten 1-[6-(2-carboxyethylsulfenyl-3-pyridine) methyl]-N-nitro-2-imidazoline imine and bovine serum albumin, preparing hybrid tumor cells from spleen cells and myeloma cells Sp2/0 of the immunized mouse by the hybrid tumor technology, and obtaining hybrid tumor strains 2F11/A9 capable of stably secreting the imidacloprid pesticide against monoclonal antibody. By effectiveness verification, the antibody can be used for sensitive and quick detection of imidacloprid residues in agricultural production environments and agricultural products. The preparation technique for the imidacloprid pesticide against monoclonal antibody is simple and feasible, does not need special instruments and equipment in the whole preparation process of the antibody, and is easy for scale production in factories.

The recombinant staphylococcus aureus enterotoxin M as one kind of superantigen with the amino acid sequence as shown in SEQ ID No. 1 is obtained through recombining gene originated from staphylococcus aureus and coding SEM with one kind of plasmid vector, transforming to proper host for expressing, and affiliation purifying. It is proved through extracorporeal mouse spleen lymphopoiesis experiment and tumor cell inhibiting experiment that the recombinant protein has typical superantigen activity stronger than that of SEC and may be applied in preparing medicine for stimulating lymphopoiesis and inhibiting tumor cell growth. The present invention is suitable for preparing high purity enterotoxin with superantigen activity and developing superantigen preparation. The present invention has reasonable design, efficient expression on recombinant SEM in the expression strength of about 15-25 % of staphylococcus aureus protein, and other advantages.

The invention discloses a Pb<2+> antigen and a corresponding monoclonal antibody, and also provides a method for preparing the same. The preparation method comprises the following steps of: chelating Pb<2+> with a chelating agent p-NH2-Bn-CHX-A''-DTPA; coupling glutaraldehyde with hemocyanin (KLH); performing ultra-filtration to obtain a full immunizing antigen (KLH-NH-Bn-CHX-A''-DTPA-Pb); merging splenic cells and myeloma cells of a mouse immunizing BALB/c; and screening a specific monoclonal antibody which has strong positive for BSA-NH-Bn-CHX-A''-DTPA-Pb and no cross reaction with BSA-NH-Bn-CHX-A''-DTPA and other various metal ions. The antibody can be used for developing ELISA kits and test paper for detecting the Pb<2+> and has great application prospect.

The invention discloses a triadimefon monoclonal antibody hybridoma cell strain B11S and application thereof, and belongs to the technical field of food safety immunodetection. The triadimefon monoclonal antibody hybridoma cell strain B11S disclosed by the invention is preserved in China General Microbiological Culture Collection Center (CGMCC), is classified and named as a monoclonal cell strain,is preserved on October 14, 2019, and has a preservation number of CGMCC No. 18518. A BALB/c mouse is immunized by a triadimefon complete antigen. High-titer and low-IC50 mouse splenocytes are takenand fused with myeloma cells through a PEG method, and the hybridoma cell strain B11S is obtained through screening of an indirect competitive enzyme-linked immunosorbent assay and three times of subcloning. A monoclonal antibody secreted by the cell strain has good specificity and detection sensitivity (IC50 value is 5.25 ng/mL) to triadimefon, provides a raw material for immunodetection of triadimefon residues in foods, and has practical application value.

The invention relates to an anti-bifenthrin monoclonal antibody and an application thereof and especially provides a monoclonal antibody capable of specifically recognizing bifenthrin, a hybrid tumor generating the monoclonal antibody and the application of the monoclonal antibody. The invention belongs to the field of biology. A method includes following steps: immunizing BALB/c female mice by an artificial antigen; performing cell fusion with myeloma cell SP2/0 and mice spleen cells to prepare a hybridoma; obtaining a hybridoma strain which can stably secret the anti-bifenthrin monoclonal antibody; injecting the hybridoma strain into abdomen of the mice to prepare ascites; purifying the ascites in an octanoic acid-ammonium sulfate manner to obtain the specific anti-bifenthrin specifically monoclonal antibody. The monoclonal antibody does not cross-react with other compounds. ELISA method established by the antibody can be used for quickly, sensitively, conveniently and cheaply detecting residual bifenthrin in environment and agricultural products.

The invention relates to a mimic eptitope peptide of BAFF-R and a DNA coding the peptide and the application of the mimic eptitope peptide in the preparation of antitumor bacterins and drugs. The mimic eptitope of 7 amino acids of BAF-R is a mimic eptitope of molecule BAFF-R with high affinity with a monoclonal antibody of BAFF-R, and is obtained through selection from a phage random display 7-peptide bank with a monoclonal antibody of BAFF-R as the antigen, wherein the sequence of the amino acids is Gly Tyr Thr Arg Trp Gly Cys. The 7-amino-acid mimic eptitope can also be used to construct a poly-peptide vaccine. The clonal inhibition rates of the phage display peptide provided by the invention are all more than 50 percent, and can specifically inhibit the combination between antibody and antigen for a larger extent, greatly improve the proliferation of mouse spleen lymphocytes without adjuvant, the mimic antigen is good in immunogenicity, as illustrated by the induced cell immune response after mice are vaccinated with the mimic antigen.

The invention discloses a polyene high-oxidation androstane compound, i.e., androstane-4, 6, 8 (9), 13 (14)-tetraene-3, 11, 16-triketone. The compound is extracted from epigynum auritum plant. Experiments prove that the compound has an obvious suppression effect on Balb/c mouse spleen lymphopoiesis stimulated by ConA (concanavalin) and has activity equivalent to that of the conventional immunosuppression active substances, i.e., triptolide and dexamethasone.

The invention discloses porcine reproductive and respiratory syndrome virus M protein CTL cell epitopes and application thereof. The identification of the epitopes comprises the following steps of: fusing and cloning a M protein gene of a porcine reproductive and respiratory syndrome virus CH-1a strain and a mouse ubiquitin gene to form a DNA vaccine and also form recombination viruses, expressing an M protein, of a WR strain vaccinia virus; immunizing a BALB/c mouse according to a Priming-Boosting policy, separating the mouse splenic lymphocyte, culturing and stimulating a CTL short peptide in vitro predicted and synthesized by a bioinformatic method, and using the flow cytometry and enzyme-linked immunospot technology to identify two CTL epitopes that are K93FITSRCRL and F57GYMTFVHF. The identification of the PRRSVM protein CTL cell epitopes lays a certain theoretical foundation for the PRRS cell immune mechanism and the novel epitope pepetide vaccine and has an instructing significance for the theoretical study on PRRS preventing and control technology.

The invention provides divaricate velvetplant root and rhizome polysaccharide and application thereof in preparing a medicine and functional food for immune adjustment and tumor resistance. According to the invention, the prepared divaricate velvetplant root and rhizome polysaccharide can promote mononuclear macrophage strain to phagocytose neutral red and release nitric oxide, can promote B cell proliferation and T cell proliferation, can also obviously improve immunosuppressive mouse spleen and thymus indexes, can improve the transformation and the proliferation of T cells and B cells, can increase the level of immunosuppressive mouse white blood cells, can obviously restrain the growth of tumor tissues, can protect organisms, can restrain the decline of weight, can gain weight, and can obviously restrain the migration of epidermic cells in veins. In addition, polysaccharide is the main functional component of divaricate velvetplant roots and rhizomes, has various health-care efficacies, is derived from purely natural materials, does not have any side effects, and can be developed into medicines and health-care products for immune adjustment, tumor resistance and angiogenesis resistance, so that the divaricate velvetplant root and rhizome polysaccharide has favorable market application prospects.

The invention discloses novel skeleton dalesconols, which has a general formula described at the right; wherein, 1,3,5,7,9,11,13,15,17,18,19,21,23,25,27 and 29 are the numbers of carbon atomic ordinal, R<1> is H or OH, when R<1> is H, dalesconols is dalesconols A; and when R<1> is OH, dalesconols is dalesconols B. The dalesconols has strong competitive inhibition to the mouse spleen cell proliferation caused by Con A, and has no toxicity to the normal mouse spleen cells; thus the dalesconols can be used as the compound with the functions of immunity and inhibition, and also can be applied to the preparation of the relevant novel medicine.