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Preparation and Characteristics Identification of Anti-kappa Casein Monoclonal Antibody

ZHANG Ying, ZHOU Yu, LI Yan-song,MENG Xing-yu,CHU Xiao-hong,LU Shi-ying,REN Hong-lin,HU Pan,LIU Zeng-shan,WANG Nan-nan,LI Dan   

  1. (Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, College of Veterinary Medicine, Jilin University, Changchun 130062, China)
  • Received:2013-09-26 Online:2014-03-20 Published:2014-05-15

Abstract: To obtain the anti-kappa casein monoclonal antibody and complete the identification of the antibody characteristics. The BALB/c mice were immunized with kappa casein using foot-pad immunization. Popliteal lymph node cells from the immunized mice were fused with SP2/0 myeloma cells in the presence of PEG. Three hybridoma strains (1C4, 3G3, 3E6) which secreted the antibody specific for kappa casein were obtained.The sub-class of the antibodies were IgG1. The ascites were purified by Protein G affinity layer absorption column. The antigenic epitope of 3G3 and 3E6 were different and it was close between 1C4 and 3E6.The titer of purified ascites(1C4) was 1.28×106 and the affinity constant was 2.89×108 mol/L. A anti-kappa casein monoclonal antibody with good affinity had been achieved,which provided foundations for the rapid detection of casein in bovine milk samples.

Key words: kappa casein; monoclonal antibody; characteristics identification