Use of a Novel Fluidics Microbead Trap/Flow-cell Enhances Speed and Sensitivity of Bead-Based Bioassays

Journal Article · Sat Sep 15 04:00:00 EDT 2007 · JALA. Journal of the Association for Laboratory Automation, 12(5):303-310

DOI:https://doi.org/10.1016/j.jala.2007.05.002· OSTI ID:921396

Ozanich, Rich M; Antolick, Kathryn C; Bruckner-Lea, Cindy J; Dockendorff, Brian P; Easterday, Ashton N; Edberg, Heather C; Grate, Jay W; Iyer, Sujata; Johnson, Laura H; Straub, Tim M; Valdez, Catherine O; Warner, Marvin G

Automated devices and methods for biological sample preparation often utilize surface functionalized microbeads (superparamagnetic or non-magnetic) to allow capture, purification and pre-concentration 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 non-magnetic functionalized beads that allow samples and reagents to be efficiently perfused over a micro-column of beads. This approach yields enhanced mass transport and up to 5-fold 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 utilizing 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 (E. coli) from 50 mL sample volumes using antibody-coupled superparamagnetic microbeads with subsequent culturing to assess capture efficiency.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 921396
• Report Number(s): PNNL-SA-54715; 24801; 400904120
• Journal Information: JALA. Journal of the Association for Laboratory Automation, 12(5):303-310, Journal Name: JALA. Journal of the Association for Laboratory Automation, 12(5):303-310 Journal Issue: 5 Vol. 12
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
DETECTION
EFFICIENCY
ESCHERICHIA COLI
Environmental Molecular Sciences Laboratory
LYMPHOKINES
NUCLEIC ACIDS
OLIGONUCLEOTIDES
POLYSTYRENE
PROTEINS
PURIFICATION
SAMPLE PREPARATION
SENSITIVITY
TRACE AMOUNTS
TRANSPORT
TRAPPING
VELOCITY
bioassay
flow-cell
fluidics
microbeads
sequential injection