Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (15): 3191-3198.doi: 10.3864/j.issn.0578-1752.2013.15.013

• STORAGE·FRESH-KEEPING·PROCESSING • Previous Articles     Next Articles

Preparation and Identification of Polyclonal Antibody of One Pen a1 Epitope Peptide

 ZHAO  Jie-1, GAO  Mei-Xu-1, PAN  Jia-Rong-2, WANG  Zhi-Dong-1, LAN  Li-Ping-1, SUI  Ke-1, XU  Shu-Ting-1, LIU  Chao-Chao-1, MOU  Hui-1   

  1. 1.中国农业科学院农产品加工研究所，北京100193
    2.中国计量学院生命科学院，杭州310018
  • Received:2013-01-29 Online:2013-08-01 Published:2013-04-02

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Abstract: 【Objective】The study was conducted to develop a new method to identify and detect the shrimp allergic protein.【Method】 Pen a 1 is the main allergic protein of shrimp, and one of its five epitope peptides was chosen and synthesized using Fmoc Method, which site in Pen a 1 was 85-105. The peptide was conjugated to keyhole limpet (KLH) and bovine serum (BSA) by glutaric dialdehyde method to get artificial immune and coating antigen, respectively. The New Zealand white rabbits were immunized with the peptide-KLH to get the antibody, and the binding ratio of artificial antigens was detected by Ellman reagent. The ic-ELISA was conducted to determine the inhibition ratio between the peptide with the antiserum. Western blot was conducted among four different protein samples to identify the specificity of the antibody to Pen a 1.【Result】The binding ratios of the artificial immune and the coating antigen were 12﹕1 and 8﹕1, respectively. The titers of the antibody reached 1.024×106, and the IC50 and the IC10 between the antibody and the peptide were 0.4324 μg•mL-1 and 0.0004 μg•mL-1, respectively. Western blot result demonstrated that the antibody could identify the allergic protein of Metapenaeus ensis (Pen a 1).【Conclusion】Antibody prepared with synthetic peptide is credible and sensitive enough for analysis and detection of targeted allergic protein.

Key words: Pen a 1 , epitope peptides , polyclonal antibodies , ic-ELISA , Western blot

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