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A capillary-based microfluidic instrument suitable for immunoaffinity chromatography.

Michael C Peoples1, Terry M Phillips, H Thomas Karnes

  • 1Department of Pharmaceutics, Virginia Commonwealth University Medical Center, P.O. Box 980533, Richmond, VA 23298-0533, USA.

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
|November 14, 2006
PubMed
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A new microfluidic system enables rapid analysis of biological samples using immunoaffinity chromatography and laser-induced fluorescence (LIF). This miniaturized device offers potential for portable, point-of-care testing with high sensitivity.

Area of Science:

  • Analytical Chemistry
  • Biotechnology
  • Microfluidics

Background:

  • Analysis of biological samples requires sensitive methods for complex matrices and small volumes.
  • Miniaturized systems offer reduced sample consumption, faster analysis, and portability for point-of-care applications.

Purpose of the Study:

  • To develop and present a prototype microfluidic system for rapid assessment of clinical samples.
  • To utilize immunoaffinity chromatography for analyte separation and laser-induced fluorescence (LIF) for detection.

Main Methods:

  • Constructed a laboratory device with syringe pumps, a nano-gradient mixing chip, micro-injector, diode laser, and PEEKsil capillary column.
  • Employed LabVIEW software for system control, step gradient elution, and LIF signal collection.

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  • Optimized the system by evaluating flow accuracy, column parameters, and pump performance.

    • Main Results:

    • Achieved a linear response (R2=0.9998) for increasing fluorescent dye concentrations via online mixing.
    • Demonstrated effectiveness of acidic gradients and confirmed detectability with a limit of detection of 10.0 pM (10.0 amol on-column).
    • Successfully demonstrated immunoaffinity chromatography using immobilized antibodies and a fluorescently labeled antigen.

    Conclusions:

    • The developed microfluidic system is capable of sensitive analyte detection and separation.
    • The system shows promise for rapid, portable, and point-of-care clinical sample analysis.
    • Immunoaffinity chromatography coupled with LIF detection is effective within this microfluidic platform.