In this paper we will describe how high affinity reagents and a sensor configuration enabling rapid mass transport can be combined for rapid, sensitive biodetection. The system that we have developed includes a renewable surface immunoassay, which involves on-column detection of a fluorescently labeled secondary antibody in a sandwich immunoassay. Yeast display and directed molecular evolution were used to create high affinity antibodies to the botulinum toxin heavy chain receptor binding domain, AR1 and 3D12. A rotating rod renewable surface microcolumn was used to form a microliter-sized column containing beads functionalized with the capture antibody (AR1). The column was perfused with sample, wash solutions, and a fluorescently labeled secondary antibody (3D12) while the on-column fluorescence was monitored. Detection was accomplished in less than 5 minutes, with a total processing time of about 10 minutes. On-column detection of botulinum toxin was more sensitive and much faster than flow cytometry analysis on microbeads using the same reagents.
Revised: January 17, 2011 |
Published: October 30, 2004
Citation
Warner M.G., B.P. Dockendorff, M.J. Feldhaus, N.C. Anheier, J.D. Marks, J.W. Grate, and C.J. Bruckner-Lea. 2004.Rapid, Sensitive Detection of Botulinum Toxin on a Flexible Microfluidics Platform. In Chemical Sensors VI: Chemical and Biological Sensors and Analytical Methods, edited by C. Bruckner-Lea, P. Vanysek, G. Hunter, et al., 2004-08, 376-380. Pennington, New Jersey:Electrochemical Society.PNNL-SA-41899.