Food Analytical Methods

, Volume 10, Issue 9, pp 3128–3136 | Cite as

Development of an icELISA and Immunochromatographic Assay for Methyl-3-Quinoxaline-2-Carboxylic Acid Residues in Fish

  • Liqiang Liu
  • Juan Peng
  • Zhengjun Xie
  • Shanshan Song
  • Hua Kuang
  • Chuanlai XuEmail author


To evaluate residues of olaquindox (OLA) and its marker compound methyl-3-quinoxaline-2-carboxylic acid (MQCA) in aquatic products in the field, five types of hapten and corresponding antigens with exposure of different antigenic determinants were designed and synthesized. The optimal immunogen-coating combinations were obtained by antiserum detection and heterologous coating screening, successfully achieving a highly specific monoclonal antibody (mAb) against MQCA. Based on the highly specific mAb 1A11 showing no cross-reactivity with other analogs, an indirect competitive ELISA and an immunochromatographic strip for MQCA detection in fish were developed using the MQCA derivative MQCA-(NH2)4 coupled to KLH as the immunogen and MQCA-NH2 coupled to OVA as the heterologous coating antigen. The IC50 value of the indirect competitive enzyme-linked immunosorbent assay (icELISA) was 3.17 ng/mL, and the linear range was 0.58–17.29 ng/mL for MQCA under optimized concentrations of coating antigen and antibody of 0.03 and 0.3 μg/mL, respectively. The recoveries of MQCA by the developed icELISA in fish samples ranged from 81.6 to 90.7%. The cutoff value of the strip was 5 ng/mL, and the limit of detection was 0.25 ng/mL under optimal working conditions of 4 μL 0.1 M K2CO3 and 50 μL 0.2 mg/mL per 1 mL colloidal gold solution. The developed icELISA and immunochromatographic strip will be very useful tools for the primary screening of MQCA in fish.


MQCA Monoclonal antibody icELISA Immunochromatographic strip 



This work is financially supported by the National Key R & D Program (2016YFD0401101) and grants from Natural Science Foundation of Jiangsu Province, MOF, and MOE (BE2016307, BK20150145, BX20151038, CMB21S1614, CLE02N1515, 201513006).

Compliance with Ethical Standards

Conflict of Interest

Juan Peng declares that she has no conflict of interests. Shanshan Song declares that she got grant from Jiangsu Province (BK20150145). Liqiang Liu declares that he got grant from Jiangsu Province (BE2016307) and MOF (201513006). Hua Kuang declares that she got grant from MOST (2016YFD0401101). Chuanlai Xu declares that he got grant from grant from Jiangsu Province and Wuxi City (BK20150145, BX20151038, CMB21S1614, CLE02N1515).

This article does not contain any studies with human subjects. All animal studies were carried out under the guidance of animal welfare committee of Jiangnan University.

Informed Consent

Not applicable.


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Liqiang Liu
    • 1
  • Juan Peng
    • 1
  • Zhengjun Xie
    • 1
  • Shanshan Song
    • 1
  • Hua Kuang
    • 1
  • Chuanlai Xu
    • 1
    Email author
  1. 1.State Key Lab of Food Science and TechnologyJiangnan UniversityWuxiChina

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