Abstract
One of the most important goals in proteomics is to detect the real-time kinetics of diverse biomolecular interactions. Fluorescence, which requires extrinsic tags, is a commonly and widely used method because of its high convenience and sensitivity. However, in order to maintain the conformational and functional integrality of biomolecules, label-free detection methods are highly under demand. We have developed the oblique-incidence reflectivity difference (OI-RD) technique for label-free, kinetic measurements of protein-biomolecule interactions. Incorporating the total internal refection geometry into the OI-RD technique, we are able to detect as low as 0.1% of a protein monolayer, and this sensitivity is comparable with other label-free techniques such as surface plasmon resonance (SPR). The unique advantage of OI-RD over SPR is no need for dielectric layers. Moreover, using a photodiode array as the detector enables multi-channel detection and also eliminates the over-time signal drift. In this paper, we demonstrate the applicability and feasibility of the OI-RD technique by measuring the kinetics of protein-protein and protein-small molecule interactions in sandwich assays.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 23307-23320 |
| Number of pages | 14 |
| Journal | Sensors (Switzerland) |
| Volume | 14 |
| Issue number | 12 |
| DOIs | |
| State | Published - 2014 |
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ASJC Scopus subject areas
- Medicine(all)
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Real-time, label-free detection of biomolecular interactions in sandwich assays by the oblique-incidence reflectivity difference technique. / Sun, Yung Shin; Zhu, Xiangdong.
In: Sensors (Switzerland), Vol. 14, No. 12, 2014, p. 23307-23320.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Real-time, label-free detection of biomolecular interactions in sandwich assays by the oblique-incidence reflectivity difference technique
AU - Sun, Yung Shin
AU - Zhu, Xiangdong
PY - 2014
Y1 - 2014
N2 - One of the most important goals in proteomics is to detect the real-time kinetics of diverse biomolecular interactions. Fluorescence, which requires extrinsic tags, is a commonly and widely used method because of its high convenience and sensitivity. However, in order to maintain the conformational and functional integrality of biomolecules, label-free detection methods are highly under demand. We have developed the oblique-incidence reflectivity difference (OI-RD) technique for label-free, kinetic measurements of protein-biomolecule interactions. Incorporating the total internal refection geometry into the OI-RD technique, we are able to detect as low as 0.1% of a protein monolayer, and this sensitivity is comparable with other label-free techniques such as surface plasmon resonance (SPR). The unique advantage of OI-RD over SPR is no need for dielectric layers. Moreover, using a photodiode array as the detector enables multi-channel detection and also eliminates the over-time signal drift. In this paper, we demonstrate the applicability and feasibility of the OI-RD technique by measuring the kinetics of protein-protein and protein-small molecule interactions in sandwich assays.
AB - One of the most important goals in proteomics is to detect the real-time kinetics of diverse biomolecular interactions. Fluorescence, which requires extrinsic tags, is a commonly and widely used method because of its high convenience and sensitivity. However, in order to maintain the conformational and functional integrality of biomolecules, label-free detection methods are highly under demand. We have developed the oblique-incidence reflectivity difference (OI-RD) technique for label-free, kinetic measurements of protein-biomolecule interactions. Incorporating the total internal refection geometry into the OI-RD technique, we are able to detect as low as 0.1% of a protein monolayer, and this sensitivity is comparable with other label-free techniques such as surface plasmon resonance (SPR). The unique advantage of OI-RD over SPR is no need for dielectric layers. Moreover, using a photodiode array as the detector enables multi-channel detection and also eliminates the over-time signal drift. In this paper, we demonstrate the applicability and feasibility of the OI-RD technique by measuring the kinetics of protein-protein and protein-small molecule interactions in sandwich assays.
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UR - http://www.scopus.com/inward/citedby.url?scp=84964313935&partnerID=8YFLogxK
U2 - 10.3390/s141223307
DO - 10.3390/s141223307
M3 - Article
C2 - 25490591
AN - SCOPUS:84924691927
VL - 14
SP - 23307
EP - 23320
JO - Sensors (Switzerland)
JF - Sensors (Switzerland)
SN - 1424-8220
IS - 12
ER -