Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 112, Issue 7, Pages 1995-1999Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1424592112
Keywords
protein immobilization; electrokinetic; microfluidics; sample preparation; protein assay
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Funding
- Defense Advanced Research Projects Agency [DARPA-ICLF10-56]
- National Institutes of Health [P01 HG000205]
- Directorate For Engineering
- Div Of Electrical, Commun & Cyber Sys [1542152] Funding Source: National Science Foundation
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The controlled immobilization of proteins on solid-state surfaces can play an important role in enhancing the sensitivity of both affinity-based biosensors and probe-free sensing platforms. Typical methods of controlling the orientation of probe proteins on a sensor surface involve surface chemistry-based techniques. Here, we present a method of tunably controlling the immobilization of proteins on a solid-state surface using electric field. We study the ability to orient molecules by immobilizing IgG molecules in micro-channels while applying lateral fields. We use atomic force microscopy to both qualitatively and quantitatively study the orientation of antibodies on glass surfaces. We apply this ability for controlled orientation to enhance the performance of affinity-based assays. As a proof of concept, we use fluorescence detection to indirectly verify the modulation of the orientation of proteins bound to the surface. We studied the interaction of fluorescently tagged anti-IgG with surface immobilized IgG controlled by electric field. Our study demonstrates that the use of electric field can result in more than 100% enhancement in signal-to-noise ratio compared with normal physical adsorption.
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