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Surface plasmon Resonance

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Surface Plasmon Resonance
K. Scott Phillips and Quan Jason Cheng

1. Introduction
SPR is an elegant surface sensitive optical technique most commonly
employed for biointeraction analysis on a flat substrate. Although the concept was suggested as early as 1968 (1,2), the use of SPR for biosensing in
its present capacity started to gain momentum from the mid-1980s (3). The
most important advantage of SPR is its label-free nature. As explained below,
SPR measures changes in the amount of material within about 200 nm of the
surface. Because detection is based on refractive index, rather than a reporter
molecule such as a fluorophore, there is no need to label the material that will
be detected. The downside of this advantage is lack of specificity. Anything
that binds or sticks to the surface will be detected, so one must be careful
to eliminate this type of interference through careful experimental design,
sophisticated surface chemistry, and often the use of a reference channel for
comparison. Another advantage of SPR is that it is conducted in real time.
Unlike endpoint measurements of binding or surface changes using bulk techniques such as fluorescence, the surface sensitivity of SPR, when used with
a properly designed flow cell, allows monitoring of a surface interaction as
it occurs. This real-time information can be fit to theoretical models to yield
kinetic and thermodynamic parameters. The result is accurate determination
of binding constants without waiting for equilibrium to be established, greatly
facilitating analysis of large compound libraries. When combined with multiplex instruments, this is especially useful for pharmaceutical screening. SPR
is also complementary to other nonlabeled surface sensitive analysis technologies such as QCM or impedance. Assuming complete monolayer coverage,
thickness changes as little as several angstroms can be detected. Finally,
because SPR is a spectroscopic technique, SPR imaging is also possible. In
the applications section, we will discuss the exciting nature of SPR imaging
spectroscopy (SPRi). For comprehensive reviews of SPR and SPR imaging,
the reader is referred to references (4–17).

From: Molecular Biomethods Handbook, 2nd Edition.
Edited by: J. M. Walker and R. Rapley © Humana Press, Totowa, NJ

809

810

K. S. Phillips and Q. Cheng

2. How SPR Works
2.1. Instrumentation
A variety of SPR instrumentation is available, ranging from inexpensive
(<$10,000 US) to extremely high-end (>$300,000). A revie...
1. Introduction
SPR is an elegant surface sensitive optical technique most commonly
employed for biointeraction analysis on a flat substrate. Although the con-
cept was suggested as early as 1968 (1,2), the use of SPR for biosensing in
its present capacity started to gain momentum from the mid-1980s (3). The
most important advantage of SPR is its label-free nature. As explained below,
SPR measures changes in the amount of material within about 200 nm of the
surface. Because detection is based on refractive index, rather than a reporter
molecule such as a fluorophore, there is no need to label the material that will
be detected. The downside of this advantage is lack of specificity. Anything
that binds or sticks to the surface will be detected, so one must be careful
to eliminate this type of interference through careful experimental design,
sophisticated surface chemistry, and often the use of a reference channel for
comparison. Another advantage of SPR is that it is conducted in real time.
Unlike endpoint measurements of binding or surface changes using bulk tech-
niques such as fluorescence, the surface sensitivity of SPR, when used with
a properly designed flow cell, allows monitoring of a surface interaction as
it occurs. This real-time information can be fit to theoretical models to yield
kinetic and thermodynamic parameters. The result is accurate determination
of binding constants without waiting for equilibrium to be established, greatly
facilitating analysis of large compound libraries. When combined with multi-
plex instruments, this is especially useful for pharmaceutical screening. SPR
is also complementary to other nonlabeled surface sensitive analysis technolo-
gies such as QCM or impedance. Assuming complete monolayer coverage,
thickness changes as little as several angstroms can be detected. Finally,
because SPR is a spectroscopic technique, SPR imaging is also possible. In
the applications section, we will discuss the exciting nature of SPR imaging
spectroscopy (SPRi). For comprehensive reviews of SPR and SPR imaging,
the reader is referred to references (4–17).
46
Surface Plasmon Resonance
K. Scott Phillips and Quan Jason Cheng
From: Molecular Biomethods Handbook, 2nd Edition.
Edited by: J. M. Walker and R. Rapley © Humana Press, Totowa, NJ
809
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