Journal Article
JALA: Journal of the Association for Laboratory Automation, vol. 12, iss. 5, pp. 303-310, 2016
Authors
Richard M. Ozanich, Kate C. Antolick, Cindy J. Bruckner-Lea, Brian P. Dockendorff, Ashton N. Easterday, Heather C. Edberg, Jay W. Grate, Sujata Iyer, Laura H. Johnson, Timothy M. Straub, Cathy O. Valdez, Marvin G. Warner
Abstract
Automated devices and methods for biological sample preparation often use surface functionalized microbeads (superparamagnetic or nonmagnetic) to allow capture, purification, and preconcentration of trace amounts of proteins, cells, or nucleic acids (DNA/RNA) from complex samples. We have developed unique methods and hardware for trapping either magnetic or nonmagnetic functionalized beads that allow samples and reagents to be efficiently perfused over a microcolumn of beads. This approach yields enhanced mass transport and up to fivefold improvements in assay sensitivity or speed, dramatically improving assay capability relative to assays conducted in more traditional “batch modes” (i.e., in tubes or microplate wells). Summary results are given that highlight the analytical performance improvements obtained for automated microbead processing systems using novel microbead trap/flow-cells for various applications including (1) simultaneous capture of multiple cytokines using an antibody-coupled polystyrene bead assay with subsequent flow cytometry detection; (2) capture of nucleic acids using oligonucleotide-coupled polystyrene beads with flow cytometry detection; and (3) capture of Escherichia coli 0157:H7 from 50-mL sample volumes using antibody-coupled superparamagnetic microbeads with subsequent culturing to assess capture efficiency.
