Bead-based assays for biodetection: from flow-cytometry to microfluidics

Richard M. Ozanich Jr.; Kathryn Antolick; Cynthia J. Bruckner-Lea; Kyle J. Bunch; Brian P. Dockendorff; Jay W. Grate; Michael A Nash; Abby Tyler; Cynthia L. Warner; Marvin G. Warner

doi:10.1117/12.819951

5 May 2009 Bead-based assays for biodetection: from flow-cytometry to microfluidics

Richard M. Ozanich Jr., Kathryn Antolick, Cynthia J. Bruckner-Lea, Kyle J. Bunch, Brian P. Dockendorff, Jay W. Grate, Michael A Nash, Abby Tyler, Cynthia L. Warner, Marvin G. Warner

Author Affiliations +

Proceedings Volume 7306, Optics and Photonics in Global Homeland Security V and Biometric Technology for Human Identification VI; 73060I (2009) https://doi.org/10.1117/12.819951
Event: SPIE Defense, Security, and Sensing, 2009, Orlando, Florida, United States

Abstract

The potential for the use of biological agents by terrorists is a real threat. Two approaches for antibody-based detection of biological species are described in this paper: 1) The use of microbead arrays for multiplexed flow cytometry detection of cytokines and botulinum neurotoxin simulant, and 2) a microfluidic platform for capture and separation of different size superparamagnetic nanoparticles followed by on-chip fluorescence detection of the sandwich complex. These approaches both involve the use of automated fluidic systems for trapping antibody-functionalized microbeads, which allows sample, assay reagents, and wash solutions to be perfused over a micro-column of beads, resulting in faster and more sensitive immunoassays. The automated fluidic approach resulted in up to five-fold improvements in immunoassay sensitivity/speed as compared to identical immunoassays performed in a typical manual batch mode. A second approach for implementing multiplexed bead-based immunoassays without using flow cytometry detection is currently under development. The goal of the microfluidic-based approach is to achieve rapid (<20 minutes), multiplexed (≥ 3 bioagents) detection using a simple and low-cost, integrated microfluidic/optical detection platform. Using fiber-optic guided laser-induced fluorescence, assay detection limits were shown to be in the 100's of picomolar range (10's of micrograms per liter) for botulinum neurotoxin simulant without any optimization of the microfluidic device or optical detection approach.

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Richard M. Ozanich Jr., Kathryn Antolick, Cynthia J. Bruckner-Lea, Kyle J. Bunch, Brian P. Dockendorff, Jay W. Grate, Michael A Nash, Abby Tyler, Cynthia L. Warner, and Marvin G. Warner "Bead-based assays for biodetection: from flow-cytometry to microfluidics", Proc. SPIE 7306, Optics and Photonics in Global Homeland Security V and Biometric Technology for Human Identification VI, 73060I (5 May 2009); https://doi.org/10.1117/12.819951

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KEYWORDS

Microfluidics

Magnetism

Flow cytometry

Luminescence

Nanoparticles

Multiplexing

Fluorescence spectroscopy

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