Jiangnan University

Hua Kuang

Xiaoling Wu

Chuanlai Xu

Liguang Xu

Liqiang Liu

Shanshan Song

Yongming Hu

Wei Ma

Aihong Wu

Wei Jiang

Lu Zeng

Maozhong Sun

Changlong Hao

Gang Wang

Libing Wang

Jie Liu

Lingling Guo

Xinxin Xu

Jianping Zhu

Lu Lin

Qiankun Zheng

Jingjing Yao

The invention discloses an anti-phenacetin monoclonal antibody hybridoma cell strain AD, a preparation method and application thereof, and relates to the technical field of food safety immunodetection. The monoclonal antibody hybridoma cell strain is named monoclonal cell strain AD and the number CGMCC19681. The Phe-BA obtained by the hydrolysis of the reaction product of the phenacetin metabolite acetaminophen and ethyl 4-bromobutyrate is used as the hapten, and the hapten is coupled with the carrier protein to prepare the immunogen Phe-BA-BSA. After the mice were immunized with the immunogen Phe-BA-BSA, they were fused with myeloma cells by PEG method, screened by indirect competitive enzyme-linked immunosorbent assay and subcloned five times to obtain hybridoma cell lines. The monoclonal antibody secreted by the cell line can be made into a phenacetin detection kit, which has good affinity and detection sensitivity for phenacetin, and can be used for immunodetection of phenacetin residues in food.

Anti-phenacetin monoclonal antibody hybridoma cell strain ad and its preparation method and application

Disclosed is a hybridoma cell strain that secretes anti-dinitolmide monoclonal antibodies applicable to the field of food safety immunoassay methods. The hybridoma cell strain DAS3H10 that secretes anti-dinitolmide monoclonal antibodies has been deposited in Comprehensive Microbiology Center of China Microbial Culture Collection Management Committee (CGMCC), addressed in No. 1 Hospital No. 3 Institute of Microbiology of the Chinese Academy of Sciences, North Chenxi Road, Beijing Chaoyang District in Beijing. It is classified as a monoclonal cell strain. The deposit date is Nov. 28, 2019, and the deposit number is MCCC No. 19165. The monoclonal antibody secreted by the hybridoma cell strain DAS3H10 has a good affinity and high sensitivity to dinitolmide. Because of ICto dinitolmide up to 9.01 ng/mL, the monoclonal antibody could be used to prepare dinitolmide immunoassay kits and colloidal gold test strips, and can further provide a powerful means for detecting dinitolmide in animal-derived foods.

Anti-dinitolmide monoclonal antibodies and hybridoma cell strain that secretes them

A hybridoma cell line of secreting cyproheptadine monoclonal antibodies with a preservation number of CGMCC No. 14699 belongs to the field of food safety immunological detection. BALB/c mice are immunized through one time immunization with complete freund's adjuvant, three times of booster immunization with incomplete freund's adjuvant and one time of rush immunization with cyproheptadine complete antigen without adjuvant; the spleen cells from BALB/C mice immunized with high potency and low value of IC50 are fused with murine myeloma cells; and then the hybridoma cell line is obtained through indirect competitive ELISA screening and three sub-clones. The monoclonal antibody secreted by this cell line has good specificity and detection sensitivity to cyproheptadine (value of IC50 is 0.37 ng/ml), being suitable for detection of cyproheptadine in food.

Hybridoma cell line of secreting cyproheptadine monoclonal antibodies and preparation method thereof

Hybridoma cell line that secrets cyproheptadine monoclonal antibodies and preparation method thereof

A hybridoma cell strain secreting a nifursol residue marker monoclonal antibody prepared in the following way: BALB/c mice are subjected to the first immunization with a complete Freund's adjuvant, subjected to booster immunization with an incomplete Freund's adjuvant for four times, and subjected to rush immunization once with nifursol residue marker complete antigen without a Freund's adjuvant so that the BALB/c mice are immunized; the spleen cells of the immunized mice with high titer and low IC50 were fused with mouse myeloma cells by a PEG method, and the fused cells are screened through indirect competitive ELISA and subcloned three times. The monoclonal antibody secreted by this cell strain has good specificity and detection sensitivity (IC50 value of 2 &#x3bc;g/L) to the nifursol residue marker and can be used for residue detection of the nifursol residual marker in food.

Hybridoma cell strain secreting nifursolol residue marker monoclonal antibody

The present disclosure discloses a lincosamides universal monoclonal antibody hybridoma cell strain and application thereof, and belongs to the technical field of food safety immunological detection. According to the present disclosure, a clindamycin chlorine-substituted derivative is used as a hapten, the hapten is coupled with bovine serum albumin (BSA) by an activated ester method to obtain an immunizing antigen, and after being uniformly mixed with a Freund's adjuvant, the immunizing antigen is subcutaneously injected to immunize BALB/c mice; clindamycin is coupled with ovalbumin (OVA) by a carbonyl diimidazole (CDI) method to be used as a coating antigen used for detecting mouse serums and a cell supernatant. The spleen cells of the immunized mice are fused with mouse myeloma cells by a PEG method, and screened by indirect ELISA and indirect competitive ELISA and subcloned three times to obtain a population-selective hybridoma cell strain. The cell strain provided by the present disclosure has relatively good inhibition on clindamycin, lincomycin and pirlimycin, and can meet the demand for lincosamides multi-residue immunoassay products on the market.

Lincosamides universal monoclonal antibody hybridoma cell strain and application thereof

A hybridoma cell strain secreting a nifursol residue marker monoclonal antibody with CGMCC No. 18525 is prepared in the following way: BALB/c mice are subjected to the first immunization with a complete Freund's adjuvant, subjected to booster immunization with an incomplete Freund's adjuvant for four times, and subjected to rush immunization once with nifursol residue marker complete antigen without a Freund's adjuvant so that the BALB/c mice are immunized; the spleen cells of the immunized mice with high titer and low IC50 were fused with mouse myeloma cells by a PEG method, and the fused cells are screened through indirect competitive ELISA and subcloned three times. The monoclonal antibody secreted by this cell strain has good specificity and detection sensitivity (IC50 value of 2 &#x3bc;g/L) to the nifursol residue marker and can be used for residue detection of the nifursol residual marker in food.

A hybridoma cell line of secreting clarithromycin monoclonal antibodies with a preservation number of hybridoma cell line of CGMCC No. 14696 belongs to the field of food safety immunological detection. BALB/c mice are immunized through one time immunization with complete freund's adjuvant, three times of booster immunization with incomplete freund's adjuvant and one time of rush immunization with clarithromycin complete antigen without adjuvant; the spleen cells from BALB/C mice immunized with high potency and low value of IC50 are fused with murine myeloma cells; and then the hybridoma cell line is obtained through indirect competitive ELISA screening and three subclones. The monoclonal antibody secreted by this cell line has good specificity and detection sensitivity to clarithromycin (value of IC50 is 0.3 ng/ml), being suitable for detection of clarithromycin in food.

Hybridoma cell line of secreting clarithromycin monoclonal antibodies and preparation method thereof

A hybridoma cell line of secreting meloxicam monoclonal antibodies with a preservation number of hybridoma cell line of CGMCC No. 14700 belongs to the field of food safety immunological detection. The. BALB/c mice are immunized through one time immunization with complete freund's adjuvant, three times of booster immunization with incomplete freund's adjuvant and one time of rush immunization with meroxicam complete antigen without adjuvant; the spleen cells from BALB/C mice immunized with high potency and low value of IC50 are fused with murine myeloma cells; and then the hybridoma cell line is obtained through indirect competitive ELISA screening and three sub-clones. The monoclonal antibody secreted by this cell line has good specificity and detection sensitivity to meloxicam (value of IC50 is 0.1 ng/ml), being suitable for detection of meroxicam in food.

Hybridoma cell line of secreting meloxicam monoclonal antibodies and application thereof

An electrochemical paper-immunosensor and method for detecting aminoglycoside antibiotics is developed in the present invention. Single-walled carbon nanotubes are coated on the common filtration paper by dip-dry cycles. With antibody against neomycin adding to the coating solution, a high sensitive biosensor for specific detection of neomycin is prepared, satisfying to the rigid authority regulations. The sensor is not only sensitive but also rapid, comparing with the classic ELISA method, with LOD of 0.04 ng mLand the whole detection process lasting less than 30 min. Another notable advantage of this invention is the versatility of the sensor, similar method is engaged to prepare the versatile sensor for other aminoglycoside antibiotics, replacement with relevant antibodies.

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Jiangsu, China

Hua Kuang