High-Throughput Nanohole Array Based System To Monitor Multiple Binding Events in Real Time
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- 作者：Jin Ji ; J. Garland O'Connell ; David J. D. Carter ; Dale N. Larson
- 刊名：Analytical Chemistry
- 出版年：2008
- 出版时间：April 1, 2008
- 年：2008
- 卷：80
- 期：7
- 页码：2491 - 2498
- 全文大小：327K
- 年卷期：v.80,no.7(April 1, 2008)
- ISSN：1520-6882
文摘
We have developed an integrated label-free, real-timesensing system that is able to monitor multiple biomolecular binding events based on the changes in theintensity of extraordinary optical transmission (EOT)through nanohole arrays. The core of the system is asensing chip containing multiple nanohole arrays embedded within an optically thick gold film, where each arrayfunctions as an independent sensor. Each array is asquare array containing 10 × 10 nanoholes (150 nm indiameter), occupying a total area of 3.3 
m × 3.3 m.The integrated system includes a laser light source, atemperature-regulated flow cell encasing the sensing chip,motorized optics, and a charge-coupled detector (CCD)camera. For demonstration purposes, sensing chipscontaining 25 nanohole arrays were studied for their usein multiplexed detection, although the sensing chip couldbe easily populated to contain up to 20 164 nanoholearrays within its 0.64 cm2 sensing area. Using thissystem, we successfully recorded 25 separate bindingcurves between glutathione S-transferase (GST) and anti-GST simultaneously in real time with good sensitivity. Thesystem responds to binding events in a concentration-dependent manner, showing a sharp linear response toanti-GST at concentrations ranging from 13 to 290 nM.The EOT intensity-based approach also enables the system to monitor multiple bindings simultaneously andcontinuously, offering a temporal resolution on milliseconds scale that is decided only by the camera speedand exposure time. The small footprint of the sensingarrays combined with the EOT intensity-based approachenables the system to resolve binding events that occurredon nanohole sensing arrays spaced 96 m apart, with areasonable prediction of resolving binding events spaced56 m apart. This work represents a new direction thatimplements nanohole arrays and EOT intensity to meethigh-throughput, spatial and temporal resolution, andsensitivity requirements in drug discovery and proteomicsstudies.
m × 3.3 m.The integrated system includes a laser light source, atemperature-regulated flow cell encasing the sensing chip,motorized optics, and a charge-coupled detector (CCD)camera. For demonstration purposes, sensing chipscontaining 25 nanohole arrays were studied for their usein multiplexed detection, although the sensing chip couldbe easily populated to contain up to 20 164 nanoholearrays within its 0.64 cm2 sensing area. Using thissystem, we successfully recorded 25 separate bindingcurves between glutathione S-transferase (GST) and anti-GST simultaneously in real time with good sensitivity. Thesystem responds to binding events in a concentration-dependent manner, showing a sharp linear response toanti-GST at concentrations ranging from 13 to 290 nM.The EOT intensity-based approach also enables the system to monitor multiple bindings simultaneously andcontinuously, offering a temporal resolution on milliseconds scale that is decided only by the camera speedand exposure time. The small footprint of the sensingarrays combined with the EOT intensity-based approachenables the system to resolve binding events that occurredon nanohole sensing arrays spaced 96 m apart, with areasonable prediction of resolving binding events spaced56 m apart. This work represents a new direction thatimplements nanohole arrays and EOT intensity to meethigh-throughput, spatial and temporal resolution, andsensitivity requirements in drug discovery and proteomicsstudies.