Synthesis of artificial antigen and ELISA identification of mouse polyclonal antiserum against clonidine hydrochloride

Authors

WANG Yao, Food and Bioengineering College, Henan University of Science and Technology, Luoyang, Henan 471003, China
CAO Jin-bo, Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou, Henan 450002, China; College of Veterinary Medicine, Northwest A&F University, Xianyang, Shaanxi 712100, China
LI Tie-mei, Chinese Academy of Inspection and Quarantine, Beijing 100176, China
PANG Xing-hao, Food and Bioengineering College, Henan University of Science and Technology, Luoyang, Henan 471003, China
HU Xiao-fei, Key Laboratory of Animal Immunology, Henan Academy of Agricultural Sciences, Zhengzhou, Henan 450002, China

Abstract

Objective: The aim of this study was to obtain highly sensitive and specific mouse polyclonal antiserum of Clonidine hydrochloride (CLO). Methods: Apraclonidine (ACLO), a derivative of CLO, was acted as hapten of clonidine (CLO) to synthesize immune antigen CLO-BSA and envelope antigen CLO-OVA by glutaraldehyde (GA). The coupling effect was identified by ultraviolet scanning and polyacrylamide gel electrophoresis. Then, BALB/c mice were immunized and their immunological characteristics were identified by indirect ELISA and indirect competitive ELISA. Results: The titer of polyclonal antiserum of all mice were higher than 1∶12 800, and the polyclonal antiserum of No.3 mouse has the best sensitivity, the IC₅₀ was 82.75 ng/mL, and the cross reaction rate with several other common clenbuterol was all less than 0.9%. Conclusion: In this study, successfully synthesized the effective artificial antigen of CLO by GA method, and obtained the mouse polyclonal antibody of CLO with high sensitivity and specificity.

Publication Date

7-28-2021

First Page

76

Last Page

80,111

DOI

10.13652/j.issn.1003-5788.2021.07.012

Recommended Citation

Yao, WANG; Jin-bo, CAO; Tie-mei, LI; Xing-hao, PANG; and Xiao-fei, HU (2021) "Synthesis of artificial antigen and ELISA identification of mouse polyclonal antiserum against clonidine hydrochloride," Food and Machinery: Vol. 37: Iss. 7, Article 12.
DOI: 10.13652/j.issn.1003-5788.2021.07.012
Available at: https://www.ifoodmm.cn/journal/vol37/iss7/12

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