864 results about "BALB/c" 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 discloses heavy chain and light chain variable regions of an anti-human BAFF monoclonal antibody. The anti-human BAFF monoclonal antibody is FMMU-BAFF-4, wherein the gene sequences of the monoclonal antibody variable regions are shown as SEQ ID NO.3 and SEQ ID NO.4; and the amino acid sequences of the monoclonal antibody variable regions are shown as SEQ ID NO.1 and SEQ ID NO.2. In the variable regions, a mouse anti-human BAFF monoclonal antibody is prepared by using a recombined human BAFF immunological BALB/c mouse; the mouse anti-human BAFF monoclonal antibody is cloned and screened to obtain a hybridoma cell line which can secrete specific human BAFF monoclonal antibody FMMU-BAFF-NO.4; the genes of the monoclonal antibody light chain and heavy chain variable regions are cloned to obtain the gene sequences and the amino acid sequences of the monoclonal antibody light and heavy chain variable regions, and CDR sequences of the variable regions; and the uniqueness of the amino acid sequences and the gene sequences is determined.

The invention discloses a hybridoma cell line of a monoclonal antibody against African swine fever virus and the secreted monoclonal antibody thereof. The preparation method of the invention comprises the following steps: preparing a recombined P30 soluble antigen by prokaryotic expression; immunizing a BALB/c mouse; and finally fusing, screening and cloning by a hybridoma technology to obtain the hybridoma cell line which can stably secrete the monoclonal antibody against African swine fever virus P30 protein. The invention further discloses a method for preparing the monoclonal antibody with the cell line, an antibody purification method and a labeling method for horseradish peroxidase of the antibody. The monoclonal antibody can be used in detecting the African swine fever viral antibody in pig serum.

The invention discloses a preparation method for a PCV-II Cap protein monoclonal antibody, an antibody and application. The invention adopts ultracentrifuged and purified PCV-II as an immunogen to immunize a BALB/c mouse by the conventional method, takes spleen cells of the immunized BALB/c mouse to fuse with SP2/0 cells, obtains two strains of hybridoma cells secreting the PCV2-Cap protein monoclonal antibodies by indirect ELISA screening, respectively names the two strains of hybridoma cells as 8-60 and 10-48, identifies biological characteristics of the two strains 8-60 and 10-48, and usesthe two strains 8-60 and 10-48 as the first antibodies to establish an indirect immunofluorescence diagnostic method. The result of the indirect immunofluorescence diagnostic method is basically consistent with that of the PCR diagnostic method, and the positive and negative coincidence rates are respectively 93.75 percent and 100 percent so as to provide reference for preventing and treating theporcine circovirus disease.

The invention discloses gene orders of schistosoma japonicum recombinant multi-epitope antigens BSjGCP-BSj23 and BSjGCP-BSj23-BSj28, a method for expressing and purifying the same, and application thereof in preparing schistosomiasis japonica immunity prevention vaccines and diagnostic reagents. Recombinant multi-epitope nucleic acid vaccines pCMV-BSjGCP-BSj23 and pCMV-BSjGCP-BSj23-BSj28 obtain 14.76 percent and 64.95 percent of worm reduction rates respectively in Kunming mice. The recombinant multi-epitope antigens pGEX-BSjGCP-BSj23 and pGEX-BSjGCP-BSj23-BSj28 obtain 15.7 percent and 57.99 percent of worm reduction rates in immunizing BalB/c mice, and obtain 91.0 percent and 89.9 percent of sensitivities as well as 97.8 percent and 93.4 percent of specificities respectively as diagnostic antigens.

The invention discloses a hybridoma cell strain capable of secreting monoclonal antibodies to quinolones and application of the monoclonal antibodies thereof. Ciprofloxacin (CIP) coupled with bovine serum albumin is used as an antigen to immunize BALB/c mice and cell fusion, screening and cloning are carried out so as to obtain one hybridoma cell strain 1F1 capable of stable passage and secretion of monoclonal antibodies (MAb) to quinolones, wherein, the accession number of the hybridoma cell strain 1F1 is CGMCC No. 5608. The titres of ascitic fluids of the 1F1 monoclonal antibodies are up to 10<-7>, and the type and the subclass of the monoclonal antibodies are IgG1 and kappa chain. According to indirect competitive ELISA analysis, the 1F1 monoclonal antibodies perform specific reactions to quinolones like ciprofloxacin, enrofloxacin, ofloxacin, danofloxacin, norfloxacin, enoxacin, marbofloxacin, sarafloxacin and difloxacin. An ELISA method, a kit and test paper for detecting residual of quinolones in food are developed by using the 1F1 monoclonal antibodies.

The invention discloses a preparing method for paclobutrazol monoclonal antibodies. The preparing method comprises the main steps as follows: (1) synthesizing artificial hapten paclobutrazol hemisuccinates by a microwave solvent free method; (2) performing coupling to obtain artificial antigens of the paclobutrazol by using the paclobutrazol hemisuccinates as raw materials; (3) performing the immune treatment on Balb/c mice by using the synthesized artificial antigens; (4) selecting the mice with optimal serum valence and optimal specificity, and mixing spleen cells and myeloma cells of the mice together in an external manner; (5) culturing and screening the fused cells by using the selective culture medium, and further cloning, propagating and storing by freezing; (7) injecting the expanded cell strains into the abdominal cavities of the mice to generate a lot of abdominal dropsy; (8) purifying the paclobutrazol-resistant monoclonal antibodies in the abdominal dropsy by a caprylic acid-saturated ammonium sulfate method. The prepared paclobutrazol-resistant monoclonal antibodies have high sensitivity and strong specificity, and can be further applied to the construction of technologies of immune sensors, colloidal gold immune chromatographic methods and the like.

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 discloses an anti-porcine reproductive and respiratory syndrome virus monoclonal antibody and application. The invention provides an anti-porcine reproductive and respiratory syndrome virus monoclonal antibody hybridoma cell strain 3C3 with collection number of CGMCC No. 4109. The invention also provides the anti-porcine reproductive and respiratory syndrome virus monoclonal antibody generated by the anti-porcine reproductive and respiratory syndrome virus monoclonal antibody hybridoma cell strain 3C3 with collection number of CGMCC No. 4109. An experiment of the invention proves that: a purified PRRSV JXA1 strain is used for immunizing BALB/c mice, one hybridoma cell strain which can stably secrete PRRSV monoclonal antibodies is screened by utilizing the hybridoma technology and the biological characteristic identification is performed on the hybridoma cell strain so as to lay the foundation for further establishing a specific, sensitive and rapid PRRSV detection method.

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 an analysis and detection technology of a human ovarian cancer tumor marker (human epididymis protein 4, HE4), in particular to a time-resolved fluorescence (TRF) immunized detection method and a kit of HE4, which is used for clinical auxiliary diagnosis, curative effect observation and prognosis judgment of the ovarian cancer. The invention comprises the following contents: constructing HE4 recombinant plasmids; expressing and purifying HE4 protein; immunizing BALB/c mice by the purified HE4 protein to prepare monoclonal antibodies; matching the obtained monoclonal antibodies to obtain two hybrid tumor cell strains (5A3 and 6C2) for secreting monoclonal antibodies of different epitopes of HE4 antigens, and marking the 5A3 monoclonal antibodies by the rare earth element Eu3+; and taking the unmarked 6C2 monoclonal antibodies as capture antibodies for coating a solid phase carrier, and taking the 5A3 monoclonal antibodies marked with the Eu3+ as detection antibodies to establish a double-antibody sandwich method for detecting HE4, thereby realizing the TRF immunized analysis of the invention.

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.

The invention relates to a kit used for rapidly detecting Escherichia coli O157:H7 in a sample, and a detection method thereof. The invention belongs to the technical field of immunological detection. According to the invention, a heated and deactivated Escherichia coli O157:H7 immunogen is used for immunizing a healthy New Zealand rabbit, such that a polyclonal antibody is obtained, and the polyclonal antibody is adopted as a coating antibody; a BALB/C mouse is immunized, and cell fusion is carried out, such that a monoclonal antibody is obtained, and the monoclonal antibody is adopted as a secondary antibody; and a double-antibody sandwich ELISA kit of Escherichia coli O157:H7 in foodstuffs (meat) is established. With the kit, a rapid and highly efficient detection means is provided for the detection of the residue of Escherichia coli O157:H7 in foodstuffs. The kit is advantaged in relatively low cost, relatively good stability, and relatively good repeatability. According to the invention, a detection limit is 105cfu/mL. The kit and the method are suitable for large-batch detections of samples.

The invention relates to a preparation method of HRPII protein monoclonal antibody of plasmodium falciparum. The preparation method comprises the following steps of: adopting HRPII protein of plasmodium falciparum as target antigen and respectively analyzing and selecting two dominant antigen epitopes of A and B; respectively repeating the two dominant antigen epitopes of A and B, then continuously connecting four glycine and forming recombinant protein C; adopting most securest code of escherichia coli and converting the amino acid sequence of the recombinant protein C into corresponding nucleotide sequence; carrying out chemical synthesis to the former step to obtain the nucleotide sequence, and respectively adding enzyme cutting sites BamHI and EcoRI at the upstream and downstream thereof; inserting nucleotide fragment obtained by the former step into expression carrier PET-28a(+), constructing recombinant protein C expression carrier and inducing to express the recombinant proteinC in the escherichia coli BL21 (DE3); carrying out ultrasonic bacteria breaking and low-temperature centrifugation, then taking supernatant of the solution, affining a chromatographic column by nickel-agarose, eluting and obtaining purified recombinant protein C; after immunizing Balb/c mouse with the recombinant protein C for a plurality of times, taking and fusing spleen cells with sp2/0 myelomacells, and obtaining six hybridoma cell lines by multiple rounds of screening; and purifying monoclonal antibody, respectively marking horse radish peroxidase and prorating matching and combination of optimum monoclonal antibody by ELISA orthogonal experiment.

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.