Production of high-affinity monoclonal antibody and development of immunoassay for 3-methyl-quinoxaline-2-carboxylic acid detection in swine muscle and liver

Xiya Zhang; Zizhe Li; Yexuan Mao; Meng Dang; Xianqing Huang; Zhanhui Wang; Huijuan Yang; Yuchen Bai; Huahai Zhang

doi:10.1016/j.foodchem.2022.135175

Production of high-affinity monoclonal antibody and development of immunoassay for 3-methyl-quinoxaline-2-carboxylic acid detection in swine muscle and liver

Food Chem. 2023 May 1:407:135175. doi: 10.1016/j.foodchem.2022.135175. Epub 2022 Dec 9.

Authors

Xiya Zhang¹, Zizhe Li¹, Yexuan Mao¹, Meng Dang¹, Xianqing Huang¹, Zhanhui Wang², Huijuan Yang³, Yuchen Bai⁴, Huahai Zhang⁵

Affiliations

¹ Henan Engineering Technology Research Center of Food Processing and Circulation Safety Control, College of Food Science and Technology, Henan Agricultural University, 63 Nongye Road, Zhengzhou, Henan 450002, China.
² Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety, China Agricultural University, Beijing 100193, China.
³ Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety, China Agricultural University, Beijing 100193, China; Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science & Nutritional Engineering, China Agricultural University, Beijing 100193, China.
⁴ School of Public Health, Shanxi Medical University, No. 56, Xinjian South Road, Taiyuan, Shanxi 030001, China. Electronic address: zhangxiya@henau.edu.cn.
⁵ College of Forestry, Northwest A&F University, Yangling, China. Electronic address: zhanghuahai@nwafu.edu.cn.

PMID: 36521388
DOI: 10.1016/j.foodchem.2022.135175

Abstract

Production of high-affinity and specific antibodies to small molecules with molecular weight (MW) lower than 200 Da is challenging. Here, we designed a novel hapten, named hapten H6, for the detection of 3-methyl-quinoxaline-2-carboxylic acid (MQCA, MW of 189 Da), a residual marker of olaquindox, one of important veterinary antibiotics. The hapten H6 maintained all structural features of MQCA, especially in mulliken atomic charge distribution. Then, a monoclonal antibody (mAb) named 8C9 was obtained with an IC₅₀ value of 0.2 µg/L, yielding a 15.5- to 88.5-fold improvement compared to previously prepared specific antibodies against MQCA. In addition, mAb 8C9 exhibited ignorable cross-reactivity with other structural analogs. Finally, a highly sensitive and specific indirect competitive ELISA based on mAb 8C9 was developed for the detection of MQCA in swine muscle and liver samples with limit of detection values of 0.04 µg/kg and 0.09 µg/kg, respectively.

Keywords: 3-Methyl-quinoxaline-2-carboxylic acid; Antibody; Hapten design; Mulliken atomic charge distribution; icELISA.

MeSH terms

Animals
Antibodies, Monoclonal* / analysis
Enzyme-Linked Immunosorbent Assay
Haptens
Immunoassay
Liver* / chemistry
Muscles / chemistry
Swine

Substances

quinoxaline-2-carboxylic acid
Antibodies, Monoclonal
Haptens