539 results about "Myeloma cell" 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 present invention provides a human antibody or an active-fragment thereof that specifically binds to human platelet membrane glycoprotein VI and does not induce a human platelet aggregation independently; a cell that produces the antibody or its active-fragment; a pharmaceutical composition that comprises the antibody or its active-fragment as an active ingredient, and so on. The above-mentioned cell can be obtained for example, as follows: a peripheral-blood-lymphocyte of the human that produces an autologous antibody to GPVI is activated by in vitro immunization under specific conditions; a hybridoma with mouse myeloma cell is prepared; and then the hybridoma that produces a monoclonal antibody, which has a binding capacity to GPVI and has an activity that suppresses collagen-mediated agglutinability of the human platelet is selected.

The invention discloses a microcystlin monoclonal antibody and the preparation method and the application thereof. The microcystlin monoclonal antibody is a microcystlin-LR monoclonal antibody, and the preparation method thereof is that: 1) an amino group is introduced to a seventh amino acid residue N-methyl dehydroalanine on microcystlin-LR, thus obtaining amino modified microcystlin-LR polypeptide; the amino modified microcystlin-LR polypeptide is coupled with carrier protein to obtain complete A antigen; 2) the immunity mouse of complete A antigen obtained in step 1) to get spleen cell of immunity mouse, which then is fused with myeloma cell of mouse to screen out positive hybrid cancer cell strain; the positive hybrid cancer cell strain is cultured or is injected in sygeneous mouse abdominal cavity to induce ascites to get the microcystlin-LR monoclonal antibody. The monoclonal antibody can be used for examining microcystlin-LR and immunity affinity purification thereof.

The invention discloses a preparation method of an anti-bisphenol A monoclonal antibody. The preparation method comprises the following steps of: combining bisphenol A and a macromolecule carrier protein to prepare an artificial immunity antigen and a coating antigen; preparing splenocyte suspension from a bisphenol A artificial immunity antigen immunity mouse; fusing the mouse splenocyte and mouse myeloma cells SP2/0 in the presence of polyethylene glycol; detecting by hybridoma technology and subcloning for many times to obtain positive hybridoma cells which can stably secrete bisphenol A antibody, wherein the secreted antibody belongs to the IgG1 subclass; and establishing a bisphenol A indirect competition ELISA (Enzyme-Linked Immunosorbent Assay) test by utilizing the monoclonal antibody secreted by cell lines with high antibodytiter, wherein the minimum limit of detection can reach 0.324ng/mL. The antibody has no cross reaction with compounds with benzene ring structures, such as benzene, phenol, tert-butylphenol, p-hydroxylphengl, o-hydroxybenzoic acid and the like, has strong specificity and can be used for immunity detection of bisphenol A.

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 present invention relates to a preparation method of heavy metal mercury monoclonal antibody, which belongs to the field of biotechnology. The present invention is particularly used in the preparation of specificity-recognition heavy metal mercury monoclonal antibody and in the fast mercury high-sensitivity measurement remained in the agricultural production and the agricultural production environment. The heavy metal mercury ion and the carrier albumen are coupled into full antigens through bifunctional metal chelate 1-(4-separate-cyano phenyl)-EDTA; the full antigens are used on Balb/C mouse and the spleen cells and the Sp2/0 myeloma cells are used to prepare hybridoma cells through the hybridoma technology. And monoclonal antibody which can stably excrete anti-Hg-EDTA is generated. The preparation technology in the present invention is easy and practical: the whole preparation process of the antigen requires no special instrument. The present invention is suitable for factory-scale production.

The invention relates to a monoclonal antibody of hepatitis B virus X protein (HBx), which is characterized by having specificity reaction to N-end epitope or C-end epitope of HBx, but having no reaction to keyhole limpet hemocyanin (KLH) of carrier protein and other expressed proteins of hepatitis B virus (HBV). The preparation method of the monoclonal antibody comprises the steps of: fusing N-end and C-end antigen epitope polypeptides of HBx respectively with mice immunized with KLH in crosslinking way, and myeloma cells and screening to obtain the monoclonal antibody. The monoclonal antibody can be used for various immunodetection reagents and directed therapeutic drugs of HBx protein or HBx antibody and applied to diagnosis and treatment of diseases such as hepatitis b virus (HBV) infection, hepatocellular carcinoma (HCC) and the like.

The invention discloses a carbendazim monoclonal antibody, a preparation method and an application thereof. The carbendazim monoclonal antibody is prepared by a mouse hybridoma cell strain MC-3 with a preservation number of CGMCC N0.4575 and the preparation method is as follows: (1) introducing ninth amino acid residue of carbendazim into one amino group to obtain the carbendazim modified by the amino and coupling the carbendazim modified by the amino with a carrier protein to obtain a complete antigen A; (2) immunizing a mouse by the complete antigen A obtained in step (1), taking splenocyteof the immune mice to fuse with a mouse myeloma cell to screen out a positive hybridoma cell strain; extensively culturing the positive hybridoma cell strain and injecting the positive cell strain into a homologous mouse abdominal cavity to induce to generate ascitic fluid and to obtain more carbendazim monoclonal antibodies. The Carbendazim monoclonal antibody can be used for detecting the carbendazim.

The invention discloses a HCMVPP65 antigenemia indirect immune fluorescence method detection kit, which uses cytomegalovirus-AD169 virus strain pp65 protein as the immunogen to immune a Balb/c mouse. The spleen cells of the immunized mouse and the myeloma cells of the mouse which belongs to the same type with the immune mouse are conventionally integrated, by indirect ELISA screening and finite dilution cloning, the hybridoma cell lines of the mouse cytomegalovirus pp65 protein cloning antibody are obtained, and the characteristics of the hybridoma cell lines are identified by ELISA, immune fluorescence experiment and other methods; two monoclonal antibodies that stably secrete the pp65 protein are established successfully and named respectively as 1A6 and 4A8. A monoclonal antibody which differs from the former report and aims at the pp65 protein of the cytomegalovirus (CMV) is prepared and a method used for preparing erythrocyte fast pyrolysis is established; compared with other detection kits which belongs to the same kind, the detection kit of the invention is faster, simpler and more convenient and has higher specificity and sensitivity.

The invention discloses a chloramphenicol universal monoclonal antibody hybridoma cell strain and application thereof, belonging to the technical field of food safety immunological detection. The preparation method comprises the following steps: by taking a thiamphenicol analogue 1-(4-aminophenyl)-2-dichlorethamin-1,3-propylene glycol as a hapten, allowing the hapten to pass through diazotization coupling bovine serum albumin (BSA), thereby obtaining a complete antigen; uniformly mixing with a Freund's adjuvant, and performing subcutaneous injection immunotherapy on BALB/c mice; synthesizing an envelope antigen to be used for screening mice serum and cell supernatant by using diazotization; and fusing splenocyte of immune mice with mouse myeloma cells through a PEG method, performing indirect ELISA and indirect competitive ELISA screening and subcloning for three times, thereby obtaining a hybridoma cell strain with mass selection property. The cell strain provided by the invention has high inhibition properties on thiamphenicol, chloramphenicol and florfenicol, and the requirements on chloramphenicol stumpy immunodetection products in the current market can be met.

An enhancer of expression, in the myeloma cell, of HM1.24 antigen, said enhancer comprising interferon-α or interferon-γ, or IRF-2 protein as an active ingredient. Interferon-α or interferon-γ is expected to enhance the expression of HM1.24 antigen by activating the promoter of a gene encoding HM1.24 antigen.

The invention relates to an anti-rabbit hemorrhagic disease virus (RHDV)VP60 albumen monoclonal antibody, and belongs to the technical field of biology. An SP2/0 myeloma cell and a BALB/c mouse splenic cell immunized by utilizing RHDV to recombine VP60 albumen undergo cell fusion, are selectively cultured by an HAT culture medium and undergo double ELISA screening by utilizing the recombined VP60 albumen and RHDV; the obtained cell culture supernatant is checked up and screened respectively to obtain a hybrid tumor cell strain A3C which can stably excrete the anti-RHDV VP60 albumen monoclonal antibody; the ascitic fluid ELISA titer of the A3C is detected to be 1:327,600; and according to identification, the monoclonal antibody can specifically combine the expressed recombined VP60 albumen as well as the RHDV, and one single reaction strip appears in both specific combinations, thereby proving that the anti-RHDV VP60 albumen monoclonal antibody is a VP60 specific antibody of the RHDV capsid albumen.

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 provides a monoclonal antibody TBCD6 resistant to mycobacterium tuberculosis culture filter protein (CFP) 10. A preparation method of the monoclonal antibody is characterized by immunizing the mice with the specific antigenic peptide, collecting sensitized B lymphocyte of the mice to undergo fusion with the myeloma cells of the mice, judging positive clones by utilizing enzyme-linked immunosorbent assay (ELISA) and western blotting, building a hybridoma cell system TBCD6 which secretes the monoclonal antibody, amplifying, culturing and injecting the positive cells to the peritoneal cavities of the mice of the same strain, using the mice to induce ascites containing the antibody and obtaining the monoclonal antibody through collection and affinity purification of the antibody. The invention provides application of the monoclonal antibody to detection and quantitative analysis of the mycobacterium tuberculosis CFP 10.

An objective of the present invention is to provide antibodies that recognize HLA class I and have significant cell death-inducing activity. To achieve the above objective, first, the present inventors immunized mice with soluble HLA-A to which a FLAG tag is attached. Four days after the final immunization, spleen cells from the mice were fused with mouse myeloma cells. Then, hybridoma screening was carried out using the cell aggregation-inducing activity and cell death-inducing activity as an index. Thus, the monoclonal antibody F17B1 that has strong cell death-inducing activity was selected. The sequences of the H chain and L chain variable regions of the mouse monoclonal antibody F17B1 were determined to humanize this antibody. Then, the humanized anti-HLA class I antibody H2-G1d_L1-k0 was assessed for cell death induction. The result showed that H2-G1d_L1-k0 suppressed the growth of the IM9 cell line in a concentration dependent manner.