Journal
REVIEW OF SCIENTIFIC INSTRUMENTS
Volume 86, Issue 11, Pages -Publisher
AMER INST PHYSICS
DOI: 10.1063/1.4934802
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Funding
- NSF [1151005]
- NIH [1R44GM106579, 1R44GM114951]
- China Scholarship Council
- Direct For Biological Sciences
- Div Of Biological Infrastructure [1151005] Funding Source: National Science Foundation
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Characterization of protein interactions is essential to the discovery of disease biomarkers, the development of diagnostic assays, and the screening for therapeutic drugs. Conventional flow-through kinetic measurements need relative large amount of sample that is not feasible for precious protein samples. We report a novel method to measure protein interaction kinetics in a single droplet with sub microliter or less volume. A droplet in a humidity-controlled environmental chamber is replacing the microfluidic channels as the reactor for the protein interaction. The binding process is monitored by a surface plasmon resonance imaging (SPRi) system. Association curves are obtained from the average SPR image intensity in the center area of the droplet. The washing step required by conventional flow-through SPR method is eliminated in the droplet method. The association and dissociation rate constants and binding affinity of an antigen-antibody interaction are obtained by global fitting of association curves at different concentrations. The result obtained by this method is accurate as validated by conventional flow-through SPR system. This droplet-based method not only allows kinetic studies for proteins with limited supply but also opens the door for high-throughput protein interaction study in a droplet-based microarray format that enables measurement of many to many interactions on a single chip. (C) 2015 AIP Publishing LLC.
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