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A Glyco-chip for the Detection of Ricin by an Automated Chemiluminescence Read-out System

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Abstract

The plant toxin ricin is a lectin that binds to D-galactose or lactose moieties by multivalent interactions. In the present work, this avidity was used to develop a novel sandwich glyco-immunoassay using a carbohydrate microarray. For realization, 6-azidohexyl-lactose was immobilized on an alkyne silane surface by Cu(I) catalyzed click chemistry. This procedure is fast, and prevents any nonspecific binding on the microarray surface. Ricin binds via its B-chain to the lactose moieties, and is detected by the biotinylated anti-ricin A-chain. By adding a horseradish peroxidase-labeled streptavidin, a chemiluminescence signal can be generated. This method is described as a sandwich-type glyco-immunoassay. The signal on the glyco-chip can be regenerated for at least 10 measurements. The limit of detection was estimated to be 80 ng mL− 1. The assay was carried out on the automated microarray readout platform MCR 3. In this way, it took 20 min for one measurement, including regeneration of the chip surface.

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Authors and Affiliations

  1. Chair of Analytical Chemistry and Institute of Hydrochemistry, Technical University of Munich, Marchioninistr. 17, Munich, Germany

    Maria Huebner, Klaus Wutz, Agathe Szkola, Reinhard Niessner & Michael Seidel

Authors
  1. Maria Huebner
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  2. Klaus Wutz
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  3. Agathe Szkola
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  4. Reinhard Niessner
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  5. Michael Seidel
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Correspondence to Michael Seidel.

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Huebner, M., Wutz, K., Szkola, A. et al. A Glyco-chip for the Detection of Ricin by an Automated Chemiluminescence Read-out System. ANAL. SCI. 29, 461–466 (2013). https://doi.org/10.2116/analsci.29.461

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  • DOI: https://doi.org/10.2116/analsci.29.461

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