
Abstract
Simultaneous detection of small and large molecules on microarray immunoassays is a challenge that limits some applications in multiplex analysis. This is the case for biosecurity, where fast, cheap and reliable simultaneous detection of proteotoxins and small toxins is needed. Two highly relevant proteotoxins, ricin (60 kDa) and bacterial toxin staphylococcal enterotoxin B (SEB, 30 kDa) and the small phycotoxin saxitoxin (STX, 0.3 kDa) are potential biological warfare agents and require an analytical tool for simultaneous detection. Proteotoxins are successfully detected by sandwich immunoassays, whereas competitive immunoassays are more suitable for small toxins (<1 kDa). Based on this need, this work provides a novel and efficient solution based on anti-idiotypic antibodies for small molecules to combine both assay principles on one microarray. The biotoxin measurements are performed on a flow-through chemiluminescence microarray platform MCR3 in 18 minutes. The chemiluminescence signal was amplified by using a poly-horseradish peroxidase complex (polyHRP), resulting in low detection limits: 2.9 +/- 3.1 small mu g L-1 for ricin{,} 0.1 +/- 0.1 small mu g L-1 for SEB and 2.3 +/- 1.7 small mu g L-1 for STX. The developed multiplex system for the three biotoxins is completely novel{,} relevant in the context of biosecurity and establishes the basis for research on anti-idiotypic antibodies for microarray immunoassays.
Item Type: | Journal article |
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Form of publication: | Publisher's Version |
Faculties: | Veterinary Medicine |
Subjects: | 500 Science > 540 Chemistry |
URN: | urn:nbn:de:bvb:19-epub-22523-4 |
Alliance/National Licence: | This publication is with permission of the rights owner freely accessible due to an Alliance licence and a national licence (funded by the DFG, German Research Foundation) respectively. |
Language: | English |
Item ID: | 22523 |
Date Deposited: | 30. Jan 2015, 13:31 |
Last Modified: | 04. Nov 2020, 13:02 |