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Competitive Immunoassays Using Antigen Microarrays

  • Zhaowei Zhang
  • Weihua Hu
  • Qi Zhang
  • Peiwu LiEmail author
  • Changming LiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1368)

Abstract

In this work, a non-fouling antigen competitive immunoassay microarray based on the polymer brush is reported to detect multiple mycotoxins. The detection is achieved by utilizing highly specific monoclonal antibodies produced in our laboratory. The polymer brush, poly[oligo(ethylene glycol) methacrylate-co-glycidyl methacrylate] (POEGMA-co-GMA), is synthesized via surface-initiated atom transfer radical polymerization (SI-ATRP) on standard glass slides. In the polymer brush, the epoxy groups of glycidyl methacrylate (GMA) residues provide covalent binding sites for spotted antigens. Moreover, the abundant poly(ethylene glycol) (PEG) side chains in the brush are able to ultimately suppress the nonspecific protein adsorption in solution (non-fouling). The polymer brush shows a high and uniform protein loading, along with a high resistance to nonspecific protein absorption that are both important to achieve a highly sensitive immunoassay. As a demonstration of a multiplex assay, aflatoxin B1 (AFB1), ochratoxin A (OTA), and zearalenone (ZEN) are selected as antigen targets for simultaneous detections using the microarray.

Key words

Competitive immunoassay Antigen microarray Mycotoxins Polymer brush 

Notes

Acknowledgement

This work was supported by the Project of National Science & Technology Pillar Plan (2012BAB19B09), the Special Fund for Agro-scientific Research in the Public Interest (201203094), and the National Natural Science Foundation of China (21205133, 31101299).

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Oil Crops Research Institute of the Chinese Academy of Agricultural SciencesWuhanPeople’s Republic of China
  2. 2.Institute for Clean Energy & Advanced MaterialsSouthwest UniversityChongqingPeople’s Republic of China

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