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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wilson DS, Nock S (2003) Recent developments in protein microarray technology. Angew Chem Int Ed 42:494–500
Seidel M, Niessner R (2014) Chemiluminescence microarrays in analytical chemistry: a critical review. Anal Bioanal Chem 406:5589–5612
Sauceda-Friebe JC, Karsunke XYZ, Vazac S, Biselli S, Niessner R, Knopp D (2011) Regenerable immuno-biochip for screening ochratoxin A in green coffee extract using an automated microarray chip reader with chemiluminescence detection. Anal Chim Acta 689:234–242
Wang Y, Liu N, Ning B, Liu M, Lv Z, Sun Z, Peng Y, Chen C, Li J, Gao Z (2012) Simultaneous and rapid detection of six different mycotoxins using an immunochip. Biosensors Bioelectron 34:44–50
de Champdore M, Bazzicalupo P, De Napoli L, Montesarchio D, Di Fabio G, Cocozza I, Parracino A, Rossi M, D’Auria S (2007) A new competitive fluorescence assay for the detection of patulin toxin. Anal Chem 79:751–757
Hu W, Liu Y, Lu Z, Li CM (2010) Poly[oligo(ethylene glycol) methacrylate-co-glycidyl methacrylate] brush substrate for sensitive surface plasmon resonance imaging protein arrays. Adv Funct Mater 20:3497–3503
Zhang D, Li P, Zhang Q, Zhang W, Huang Y, Ding X, Jiang J (2009) Production of ultrasensitive generic monoclonal antibodies against major aflatoxins using a modified two-step screening procedure. Anal Chim Acta 636:63–69
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).
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Zhang, Z., Hu, W., Zhang, Q., Li, P., Li, C. (2016). Competitive Immunoassays Using Antigen Microarrays. In: Li, P., Sedighi, A., Wang, L. (eds) Microarray Technology. Methods in Molecular Biology, vol 1368. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3136-1_17
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3136-1_17
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3135-4
Online ISBN: 978-1-4939-3136-1
eBook Packages: Springer Protocols