464 results about "BALB/c Mouse" patented technology

The invention discloses a subunit coronavirus vaccine for dimerization-based receptor binding domains and belongs to the technical field of medicine. A baculovirus expresses RBD (receptor binding domain (E367-Y606) of MERS-CoV (middle east respiratory syndrome coronavirus) protein and RBD (R294-F515) of SARS-CoV (severe acute respiratory syndrome coronavirus) in insect cells, the RBDs may form a dimer through cysteine residue at 603 of S (spike) protein or form a dimer through cysteine residue at 512 of the S protein, and purified RBD protein dimer and monomer are used respectively to immunize Bald/c mice. The dimerized RBDs have the advantages that the defect that RBD monomers have poor immunogenicity is overcome and the generation of neutralizing antibodies in against MERS-CoV is increased greatly.

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 a kit for rapid detection of staphylococcus aureus in a sample and a detection method thereof, belonging to the technical field of immunological detection. In the invention, a staphylococcus aureus immunogen inactivated with formaldehyde is used for immunizing a healthy New Zealand rabbit to obtain a polyclonal antibody to serve as a coated antibody, and used for immunizing a BALB/C mouse and performing cell fusion to obtain a monoclonal antibody to serve as a secondary antibody, thus, the kit for performing a double antibody sandwich enzyme-linked immunosorbent assay on the staphylococcus aureus in food (milk) is established, and a rapid and efficient detection means is provided for residual detection of the staphylococcus aureus in the food, and the advantages of lower cost and better stability and repeatability are achieved. A detection limit of the kit is 105cfu/mL and is suitable for detecting mass samples.

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 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 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.

The invention discloses variable regions of light chains and heavy chains of FMU-EPCAM-2A9 monoclonal antibodies. The amino acid sequence of a monoclonal antibody light chain variable region is shown as SEQ ID NO.1, and the amino acid sequence of a monoclonal antibody heavy chain variable region is shown as SEQ ID NO.2. A set of mouse anti-human EPCAM monoclonal antibodies are prepared through recombining human EPCAM immune BALB/c mice, and hybridoma cell lines capable of stably secreting high-affinity anti-human EPCAM monoclonal antibodies are screened to prepare ascitic fluid so as to obtain the high-affinity anti-human EPCAM monoclonal antibodies. The uniqueness of the gene sequence and a corresponding protein sequence and a CDR sequence of the gene sequence are confirmed so as to provide supports for anti-human EPCAM chimeric or humanized gene engineering antibodies.

The present invention is the preparation process and application of one monoclonal antibody. The monoclonal antibody of the present invention is monoclonal antibody of Staphylococcus aureus coat protein. The preparation process includes immunizing Balb/C mouse with sterilized Staphylococcus aureus, fusing the spleen cell and Sp2/0 myelomatosis cell, and screening and cloning to obtain myelomatosis strain secreting Staphylococcus aureus resisting monoclonal antibody and the monoclonal antibody. The monoclonal antibody of the present invention is used in preparing ELISA kit or detecting colloidal gold kit.

The invention relates to a double-antibody sandwich method for detecting enterobacter sakazakii in food. The method is characterized in that the enterobacter sakazakii (ATCC29544) and lipopolysaccharide (LPS) extracted by coupling the enterobacter sakazakii (ATCC29544) with BSA (bovine serum albumin) are respectively used as immunogen to simultaneously immunize BALB/c mice, and 11 enterobacter sakazakii monoclonal antibodies are obtained through immunization, fusion and screening to respectively label HRP (horse radish peroxidase) and are paired through the enterobacter sakazakii. A No.7 monoclonal antibody CGMCC No. 7207 is used as a coating antibody, a No.8 monoclonal antibody CGMCCNO.7208 is used as an enzyme labeled antibody, a sandwich ELISA method of the enterobacter sakazakii is established by adopting the ATCC29544 as a standard product, and LOD is 53000CFU/Ml. By adopting the monoclonal antibody with high physicochemical property, good uniformity, good specificity and capability of being in mass preparation, the established sandwich method is high in sensitivity and low in cost, no cross reaction with salmonella enteritidis, E.coli, E.coliO157: H7, salmonella typhimurium, staphylococcus aureus and listeria monocytogenes exists, and thus a rapid and high efficient analysis means is provided for detecting the enterobacter sakazakii in the food.

The invention discloses preparation of thiacloprid monoclonal antibodies. The preparation method comprises the steps of synthesizing thiacloprid haptens, coupling the thiacloprid haptens with bovine serum albumin to prepare immunizing antigens, immunizing a BALB/c mouse, fusing the spleen cells of the immunized mouse with Sp2/0 myeloma cells, and performing screening and cloning to obtain hybrid strains of continuous and stable secreting thiacloprid monoclonal antibodies. The thiacloprid monoclonal antibodies have high affinity to thiacloprid and strong specificity, and do not have cross reaction with nicotine insecticide similar to acetamiprid, imidacloprid and the like.

The invention discloses a universal phthalic acid esters monoclonal antibody hybridoma cell strain and application thereof, and belongs to the technical field of food safety immunological detection. Hapten is prepared and is coupled with albumen based on the glutaraldehyde method to obtain phthalic acid dibutyl ester complete antigen, and the phthalic acid dibutyl ester complete antigen and freund's adjuvant are evenly mixed to be injected to immune BALB/c mice in a subcutaneous injection mode; envelope antigen is formed in a synthesis mode based on the diazotization method and is used for screening mouse serum and cell supernatant. The splenocyte of the immune mice is fused with myeloma cells of the mice based on a PEG method, and indirect ELISA, indirect competition ELISA screening and three times of subcloning are carried out to obtain selective group hybridoma cell strain monoclonal cell strain C. The monoclonal cell strain C has certain recognition capability on DEHP and DINP, and the requirement for phthalic acid ester plasticizer immunodetection products in current market can be met.