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Label-free molecular interaction determinations with nanoscale interferometry.

Dmitry A Markov1, Kelly Swinney, Darryl J Bornhop

  • 1Department of Chemistry, Vanderbilt University, VU Station B 351822, Nashville, Tennessee 37235-1822, USA.

Journal of the American Chemical Society
|December 17, 2004
PubMed
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This study introduces backscattering interferometry in rectangular channels (BIRC) for label-free quantification of molecular interactions in picoliter volumes. BIRC enables sensitive, noninvasive detection of binding events, advancing cellular function and therapeutic studies.

Area of Science:

  • Biophysics
  • Analytical Chemistry
  • Microfluidics

Background:

  • Quantifying protein-protein and ligand-substrate interactions is crucial for cellular biology and drug development.
  • Label-free assays are preferred for mimicking biological conditions but face challenges in microfluidic integration and sensitivity at low volumes.
  • Existing methods struggle to reduce detection volumes for universal analyte quantification without compromising sensitivity.

Purpose of the Study:

  • To develop a method for label-free quantification of molecular interactions in picoliter volumes within microfluidic devices.
  • To demonstrate the capability of backscattering interferometry in rectangular channels (BIRC) for sensitive and noninvasive detection.
  • To overcome limitations of existing label-free assay tools for microfluidic applications.

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Main Methods:

  • Utilized backscattering interferometry in rectangular microfluidic channels (BIRC) molded in poly(dimethylsiloxane) (PDMS).
  • Employed low-power coherent radiation for optical detection within a 225 pL detection volume.
  • Quantified both irreversible streptavidin-biotin binding and reversible protein A-human IgG Fc interactions.

Main Results:

  • Achieved detection limits of 47 x 10^-15 mol for biotin and 2 x 10^-15 mol for IgG Fc.
  • Demonstrated high-sensitivity universal solute quantification using interferometry in microfluidic channels.
  • Successfully performed label-free quantification of reversible molecular interactions with femtomole sensitivity.

Conclusions:

  • BIRC facilitates label-free molecular interaction studies in picoliter volumes with high sensitivity.
  • The method is compatible with inexpensive PDMS microfluidic chips and low-power optics.
  • BIRC shows potential for high-throughput binding affinity quantification in microfluidic systems.