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Clinical Chemistry 51: 1962-1972, 2005. First published August 4, 2005; 10.1373/clinchem.2005.053249
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(Clinical Chemistry. 2005;51:1962-1972.)
© 2005 American Association for Clinical Chemistry, Inc.

Oak Ridge Conference

Direct Quantification of Analyte Concentration by Resonant Acoustic Profiling

Benjamin Godber, Kevin S.J. Thompson, Marian Rehak, Yildiz Uludag, Sven Kelling, Alexander Sleptsov, Mark Frogley, Klaus Wiehler, Christopher Whalen and Matthew A. Coopera

1 Akubio Ltd., Cambridge, United Kingdom.

aAddress correspondence to this author at: Akubio Ltd., Unit 181, Cambridge Science Park, Cambridge CB4 0GJ, UK. Fax 44-1223-225336; e-mail mcooper{at}akubio.com.


Abstract

Background: Acoustic sensors that exploit resonating quartz crystals directly detect the binding of an analyte to a receptor. Applications include detection of bacteria, viruses, and oligonucleotides and measurement of myoglobin, interleukin 1ß (IL-1ß), and enzyme cofactors.

Methods: Resonant Acoustic ProfilingTM was combined with a microfluidic lateral flow device incorporating an internal reference control, stable linker chemistry, and immobilized receptors on a disposable sensor "chip". Analyte concentrations were determined by analyzing the rate of binding of the analyte to an appropriate receptor.

Results: The specificity and affinity of antibody–antigen and enzyme–cofactor interactions were determined without labeling of the receptor or the analyte. We measured protein concentrations (recombinant human IL-1ß and recombinant human myoglobin) and quantified binding of cofactors (NADP+ and NAD+) to the enzyme glucose dehydrogenase. Lower limits of detection were ~1 nmol/L (17 ng/mL) for both IL-1ß and human myoglobin. The equilibrium binding constant for NADP+ binding to glucose dehydrogenase was 2.8 mmol/L.

Conclusions: Resonant Acoustic Profiling detects analytes in a relatively simple receptor-binding assay in <10 min. Potential applications include real-time immunoassays and biomarker detection. Combination of this technology platform with existing technologies for concentration and presentation of analytes may lead to simple, label-free, high-sensitivity methodologies for reagent and assay validation in clinical chemistry and, ultimately, for real-time in vitro diagnostics.




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Rapid and Simultaneous Quantification of 4 Urinary Proteins by Piezoelectric Quartz Crystal Microbalance Immunosensor Array
Clin. Chem., December 1, 2006; 52(12): 2273 - 2280.
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L. Huang and M. A. Cooper
Real-time label-free acoustic technology for rapid detection of Escherichia coli O157:H7.
Clin. Chem., November 1, 2006; 52(11): 2148 - 2151.
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