Four Hapten Spacer Sites Modulating Class Specificity: Nondirectional Multianalyte Immunoassay for 31 β-Agonists and Analogues

  • Anal Chem. 2018 Feb 20;90(4):2716-2724. doi: 10.1021/acs.analchem.7b04684.
Lanteng Wang  1 ,  Wenmeng Jiang  1 ,  Xing Shen  1 ,  Xiangmei Li  1 ,  Xin-An Huang  2 ,  Zhenlin Xu  1 ,  Yuanming Sun  1 ,  Shun-Wan Chan  3 ,  Lingwen Zeng  4 ,  Sergei Alexandrovich Eremin  5  6 ,  Hongtao Lei  1
Affiliations
  • 1. Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University , Guangzhou 510642, China.
  • 2. Tropical Medicine Institute & South China Chinese Medicine Collaborative Innovation Center, Guangzhou University of Chinese Medicine , Guangzhou 510405, China.
  • 3. Faculty of Science & Technology, Technology & Higher Education Institute of Hong Kong , Hong Kong, China.
  • 4. South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences , Guangzhou 510530, China.
  • 5. Faculty of Chemistry, M.V. Lomonosov Moscow State University , 119991 Moscow, Russia.
  • 6. A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences , 119071 Moscow, Russia.
PMID: 29353474 DOI: 10.1021/acs.analchem.7b04684
Abstract

Immunoassay methods are important for monitoring β-agonists illegally used for reducing animal fat deposition in livestock. However, there is no simultaneous screening surveillance Immunoassay for detecting various β-agonist chemicals that are possibly present in food. In this study, through the use of an R-(-)-salbutamol derivative as the immunizing hapten, an antibody recognizing 31 β-agonists and analogues was generated for the first time. Three-dimensional quantitative structure-activity relationship (3D QSAR) revealed that strong steric and hydrophobic fields around the hapten spacer near C-2, as well as a chirality at C-1', dominantly modulated the class specificity of the raised antibody. However, a hapten spacer linked at C-2' or C-1 would lead to a narrow specificity, and the spacer charge at C-6 could affect the raised antibody specificity spectrum. A class specificity competitive indirect enzyme-linked immunosorbent assay (ciELISA) was established with an ideal recovery ranging from 81.8 to 118.3% based on the obtained antibody. With a good agreement to the HPLC/MS method, the proposed ciELISA was confirmed to be reliable for the rapid surveillance screening assay of β-agonists in urine. This investigation will contribute to the rational design and control of the Immunoassay specificity.

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