FOOD SCIENCE ›› 2013, Vol. 34 ›› Issue (5): 181-185.doi: 10.7506/spkx1002-6630-201305038

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Preparation and Identification of Streptomycin Artificial Antigen and Its Polyclonal Antibody

XI Xi1,LI Mu-jie1,GONG Yun-fei1,CHEN Zong-lun2,FANG Jie-hong1,WANG Wei-fen1,ZHANG Ming-zhou1,*   

  1. 1. Zhejiang Provincial Key Laboratory of Biometrology, Inspection and Quarantine, College of Life Science, China Jiliang University,
    Hangzhou 310018, China;2. Hangzhou DNA Scitech Co. Ltd., Hangzhou 310013, China
  • Received:2011-12-18 Revised:2013-01-21 Online:2013-03-15 Published:2013-04-16
  • Contact: ZHANG Ming-zhou E-mail:zmzcjlu@cjlu.edu.cn

Abstract:

In order to develop an immunoassay for treptomycin (SM) residues, the synthesis of antigen and preparation of
polyclonal antibody for SM were explored. In this study, immunogen SM-cBSA was synthesized by EDC after inserting
carboxy group by O-carboxymethyl-hydroxylamine. Coating antigen SM-OVA was synthesized by glutaraldehyde.
Both SM-cBSA and SM-OVA were identified with UV-visible spectroscopy and SDS-PAGE to reveal the successful
coupling with a molar coupling ratio of 7.6:1 and 17.7:1, respectively. During animal immunization, the antibody
titer and IC50 were detected by indirect ELISA and measured by competitive ELISA to be 1:40000 and 3.32 ng/mL,
respectively. Meanwhile, the cross-reactivity of streptomycin polyclonal antibody with dihydrostreptomycin was 105.21%.

Key words: streptomycin, artificial antigen, polyclonal antibody, indirect ELISA

CLC Number: