Surface Plasmon Resonance imaging (SPRi) is a label-free technique for the quantitation of binding affinities and
concentrations for a wide variety of target molecules. Although SPRi is capable of determining binding constants for
multiple ligands in parallel, current commercial instruments are limited to a single analyte stream and a limited number
of ligand spots. We have developed an integrated microfluidic array using soft lithography techniques for SPRi-based
detection and determination of binding affinities for DNA aptamers against human alpha-thrombin. The device consists
of 264 element-addressable chambers of 700 pL each isolated by microvalves. The device also contains a dilution
network for simultaneous interrogation of up to six different target concentrations, further speeding detection times. The
element-addressable design of the array allows interrogation of multiple ligands against multiple targets, and analytes
from individual chambers may be collected for downstream analysis. We demonstrate methods for educing nonspecific
binding to the sensor surface and quantify the success of these methods using mass spectrometric identification of
proteins eluted from our microfluidic chambers following SPRi analysis of crude cell lysates.
Surface Plasmon Resonance imaging (SPRi) is a label-free technique for the quantitation of binding affinities and
concentrations for a wide variety of target molecules. Although SPRi is capable of determining binding constants for
multiple ligands in parallel, current commercial instruments are limited to a single analyte stream and a limited number
of ligand spots. Measurement of target concentration also requires the serial introduction of different target
concentrations; such repeated experiments are conducted manually and are therefore time-intensive. Likewise, the
equilibrium determination of concentration for known binding affinity requires long times due to diffusion-limited
kinetics to a surface-immobilized ligand. We have developed an integrated microfluidic array using soft lithography
techniques for SPRi-based detection and determination of binding affinities for DNA aptamers against human alphathrombin.
The device consists of 264 element-addressable chambers of 700 pL each isolated by microvalves. The device
also contains a dilution network for simultaneous interrogation of up to six different target concentrations, further
speeding detection times. The element-addressable design of the array allows interrogation of multiple ligands against
multiple targets, and analytes from individual chambers may be collected for downstream analysis.
Surface Plasmon Resonance imaging (SPRi) is a label-free technique for the quantitation of binding affinities and
concentrations for a wide variety of target molecules. Although SPRi is capable of determining binding constants for
multiple ligands in parallel, current commercial instruments are limited to a single analyte stream and a limited number
of ligand spots. Measurement of target concentration also requires the serial introduction of different target
concentrations; such repeated experiments are conducted manually and are therefore time-intensive. Likewise, the
equilibrium determination of concentration for known binding affinity requires long times due to diffusion-limited
kinetics to a surface-immobilized ligand. We have developed an integrated microfluidic array using soft lithography
techniques for SPRi-based detection and determination of binding affinities for DNA aptamers against human alphathrombin.
The device consists of 264 element-addressable chambers isolated by microvalves. The resulting 700 pL
volumes surrounding each ligand spot promise to decrease measurement time through reaction rate-limited kinetics. The
device also contains a dilution network for simultaneous interrogation of up to six different target concentrations, further
speeding detection times. Finally, the element-addressable design of the array allows interrogation of multiple ligands
against multiple targets.
Conference Committee Involvement (5)
Biosensing and Nanomedicine II
12 August 2012 | San Diego, California, United States
Biosensing and Nanomedicine
21 August 2011 | San Diego, California, United States
Biosensing III
1 August 2010 | San Diego, California, United States
Biosensing II
4 August 2009 | San Diego, California, United States
Biosensing
12 August 2008 | San Diego, California, United States
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