Anal. Chem.

2008, 80, 7174–7178

Aptamer-Based Au Nanoparticles-Enhanced
Surface Plasmon Resonance Detection of Small
Molecules
Jianlong Wang and H. Susan Zhou*

Department of Chemical Engineering, Worcester Polytechnic Institute, 100 Institute Road, Worcester, Massachusetts 01609

Small molecules are difficult to detect by conventional label-free, high-selectivity, and high-sensitivity assay method.2
SPR technique directly because the changes in the refrac- However, these researches mainly focus on studying the binding
tive index resulting from the binding processes of small processes between large biomolecules; small molecules are
biomolecules are often small. In order to extend the seldom detected by conventional SPR technique directly because
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application of SPR biosensor in detecting a small mol-

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the changes in the refractive index resulting from the binding

ecule, we combine the advantage of aptamer technique
processes of small biomolecules are often small. In order to extend
with the amplifying effect of Au nanoparticles to design a
the application of SPR biosensor and develop a novel SPR sensor
sensitive SPR sensor for detecting small molecules. The
for detection of general small molecules with high sensitivity and
principle of this sensor is based on surface inhibition
detection. The aptamer is first immobilized on SPR gold selectivity, two issues need to be considered: (1) how to simply
film with its ss-DNA structure. The aptamer possessing construct a recognition surface capable of fast and reliable
this structure can be hybridized with Au nanoparticles- interaction with small molecules; (2) how to amplify the change
tagged complementary ss-DNA and result in a large of SPR signal resulting from the binding of small molecules with
change of SPR signal. However, the aptamer will change the ligands immobilized on the SPR sensor surface.
its structure from ss-DNA to tertiary structure after The occurrence of an aptamer technique provides an op-
adenosine is added to the SPR cell. The aptamer pos- portunity to fabricate a sensing surface for simple and effective
sessing tertiary structure could not hybridize with Au detection of small molecules by SPR spectroscopy. Numerous
nanoparticles-tagged complementary ss-DNA. Thus, the reports had confirmed that aptamers can specifically respond to
change of SPR signal resulted in the hybridization reaction all kinds of small molecules that can be obtained by SELEX.3
between aptamer and Au nanoparticles-tagged comple-
Furthmore, aptamers can be easily labeled with mercaptan during
mentary ss-DNA will decrease with the increase of the
their synthetic processes,4 which enable the immobilization of
number of aptamers possessing tertiary structure, which
aptamer on the surface of SPR gold film for detection of small
is proportional to the concentration of the small molecule.
Based on this principle, we choose a simple system molecules. Comparing with other assembling methods that always
(antiadenosine aptamer/adenosine) to detect the sensing need several steps ((1) immobilize functional mercaptan; (2) attach
ability of this SPR biosensor for a small molecule. The protein or other biomolecules) to construct a sensing surface,
experimental results confirm that the SPR sensor we aptamer can be directly immobilized on a gold surface, which
developed possesses a good sensitivity and a high selec- greatly simplifies the procedure of the experiments. However, very
tivity for adenosine. The detection range for adenosine is few reports have appeared on developing an SPR biosensor for
from 1 × 10-9 to 1 × 10-6 M. More significantly, it is
fairly easy to generalize this strategy to detect a spectrum
(2) (a) Deckert, F.; Legay, F. Anal. Biochem. 1999, 274, 81–89. (b) Shankaran,
of small molecules by SPR spectroscopy using different D. R.; Gobi, K. V. A.; Miura, N. Sens. Actuators, B 2007, 121, 158–177. (c)
aptamers. Therefore, it is expected that this method may Vutukuru, S. R.; Bethi, R. S.; Kane, R. S. Langmuir 2006, 22, 10152. (d)
offer a new direction in designing high-performance SPR Dong, Y.; Phillips, K. S.; Cheng, Q. Lab Chip 2006, 6, 675–681. (e)
Georgiadis, R.; Peterling, K. P.; Peterson, A. W. J. Am. Chem. Soc. 2000,
biosensors for sensitive and selective detection of a wide 122, 3166–3173.
spectrum of small molecules. (3) (a) Navani, N. K.; Li, Y. Curr. Opin. Chem. Biol. 2006, 10, 272–281. (b)
Willner, I.; Zayats, M. Angew. Chem., Int. Ed. 2007, 46, 6408–6418. (c)
Lin, C.; Katilius, E.; Liu, Y.; Zhang, J.; Yan, H. Angew. Chem., Int. Ed. 2006,
Optical surface plasmon resonance (SPR) spectroscopy is a 45, 5296–5301. (d) Nutiu, R.; Li, Y. Methods 2005, 37, 16–25. (e) Nutiu,
powerful tool for in situ real-time characterization of solid/liquid R.; Yu, J. M.; Li, Y. ChemBioChem 2004, 5, 1139–1144. (f) Nutiu, R.; Li, Y.
Chem.-Eur. J. 2004, 10, 1868–1876. (g) Nutiu, R.; Li, Y. J. Am. Chem. Soc.
interfaces.1 This technology has been widely used for the study 2003, 125, 4771–4778. (h) Rupcich, N.; Chiuman, W.; Nutiu, R.; Mei, S.;
of interactions of biological molecules because it provides a rapid, Flora, K. K.; Li, Y.; Brennan, J. D. J. Am. Chem. Soc. 2006, 128, 780–790.
(4) (a) Liss, M.; Petersen, B.; Wolf, H.; Prohaska, E. Anal. Chem. 2002, 74,
* To whom correspondence should be addressed. Tel.:508-831-5275, Fax: 508- 4488–4495. (b) Hendrix, M.; Priestley, E. S.; Joyce, G. F.; Wong, C. H. J. Am.
831-5936. E-mail: [email protected]. Chem. Soc. 1997, 119, 3641–3648. (c) Verhelst, S. H. L.; Michiels, P. J. A.;
(1) Kang, X. F.; Jin, Y. D.; Cheng, G. J.; Dong, S. J. Langmuir 2002, 18, 1713– an der Marel, G. A.; van Boeckel, C. A. A.; van Boom, J. H. ChemBioChem
1718. 2004, 5, 937–942.

7174 Analytical Chemistry, Vol. 80, No. 18, September 15, 2008 10.1021/ac801281c CCC: $40.75  2008 American Chemical Society
Published on Web 08/16/2008
small molecules based on the aptamer technique.5 In this paper, HNO3 (aqua regia) and rinsed thoroughly in H2O prior to use.
we apply aptamer technique to SPR and develop a general SPR Au nanoparticles stabilized with citrate were synthesized according
biosensor to detect small molecules. As for the second problem, to the literature procedure.10 That is, 100 mL of 1 mM HAuCl4 (4
the electronic coupling interaction between the localized surface mL of 1% (w/w) HAuCl4 solution dissolved in 96 mL of H2O) was
plasmon of the Au nanoparticles and the surface plasmon wave brought to a reflux while stirring and then 10 mL of a 38.8 mM
associated with the SPR gold film had been demonstrated to be trisodium citrate (10 mL of 1.14% (w/w) trisodium citrate) solution
able to greatly enhance the signal of SPR.6 Many Au nanoparticles- was added quickly, which resulted in a color change of the solution
enhanced SPR biosensors had been developed for the detection from pale yellow to deep red. After the color change, the solution
of large biomolecules, such as DNA and protein.7 However, was refluxed for an additional 15 min and left to cool to room
reports on using this method to detect small molecules are still temperature. The UV-vis spectrum shows the maximum extinc-
limited.8 Currently the existing SPR biosensor for small molecules tion value of the 519-nm plasmon peak is ∼3.0 as shown in Figure
is mainly constructed by an indirect competitive immunoassay S1 (Supporting Information) curve a.
method.9 This method uses tedious steps to construct the sensing Au nanoparticles modified by ss-DNA were prepared according
surface. Furthermore, small molecules coexisted in solution with to the literature with some modification.11 That is, transfer 3 mL
a large amount of antibody proteins, which easily affects the of the already prepared Au nanoparticles to the NaOH-treated
accuracy of the detection results. Thus, it is imperative to develop glass vials and then add 60 µL of 5 µM ss-DNA with magnetic
novel methods that can simply and directly detect small molecules stirring to facilitate the reaction for 16 h. The final Tris-HCl
by SPR spectroscopy. concentration is ∼1 mM. Centrifuge the modified Au nanoparticles
Here, we combine the amplifying characteristic of Au nano- at 14000g at room temperature for 25 min twice to remove the
particles with the advantage of aptamer technique to design a free ss-DNA. At last, disperse the Au nanoparticles in 2 mL of
“pseudo” sandwich reaction for detecting small molecules by SPR buffer containing 2.8 mM NaCl, 1 mM Tris-HCl, pH 8.0.
spectroscopy. UV-vis absorption spectra of Au nanoparticles modified by
ss-DNA before centrifugation are shown in curve b of Figure S1
EXPERIMENTAL SECTIONS
(Supporting Information), from which we can see that the addition
Materials. Trisodium citrate, hydrogen tetrachloroaurate(III)
of ss-DNA into Au nanoparticles stabilized by citrate did not
(HAuCl4), 6-mercaptohexan-1-ol, adenosine, uridine, cytidine, and
change the absorbance of Au nanoparticles at 519 nm greatly.
guanosine were purchased from Sigma and used as received. DNA
After centrifugation, a visible absorption peak appears at 519.5
molecules were obtained from Integrated DNA Technologies
nm (curve c) with a decreased intensity. The decrease of peak
(IDT). The sequence of the adenosine-binding aptamer was 5′-
intensity mainly comes from the loss of Au nanoparticles during
SH-C6-AGA GAA CCT GGG GGA GTA TTG CGG AGG AAG GT-
the process of centrifugation.
3′ (aptamer); the sequence of its part complementary strand was
In Situ SPR Measurement. The SPR experiments were done
5′-SH-C6-ACC TTC CTC CGC-3′ (ss-DNA). DNA solutions were
using Eco Chemie Autolab SPR systems (Brinkmann Instruments,
prepared by dissolving DNA in 50 mM pH 8.0 Tris-HCl buffer
New York). It works with a laser diode fixed at a wavelength of
including 138 mM NaCl. Different concentrations of adenosine
670 nm, using a vibrating mirror to modulate the angle of
and 1 mM uridine, cytidine, and guanosine were all prepared in
incidence of the p-polarized light beam on the SPR substrate. The
the Tris-HCl buffer.
instrument is equipped with a cuvette. A gold sensor disk (25 mm
Synthesis and Modification of Au Nanoparticles with a
in diameter) was mounted on the hemicylindrical lens (with index-
Diameter of ∼13 nm. All glassware used in the following
matching oil) to form the base of the cuvette. An O-ring (3-mm
procedures were cleaned in a bath of freshly prepared 3:1 HCl/
inner diameter) between the cuvette and disk prevents leakage.
(5) (a) Win, M. N.; Klein, J. S.; Smolke, C. D. Nucleic Acids Res. 2006, 19, An autosampler (Eco Chemie) with controllable aspirating-
5670–5682. (b) Redmen, J. E. Methods 2007, 43, 302–312. (c) Seenisamy, dispensing-mixing pipet was used to add samples into the cuvette
J.; Bashyam, S.; Gokhale, V.; Vankayalapati, H.; Sun, D.; Siddiqui-Jain, A.;
Streiner, N.; Shin-ya, K.; White, E.; Wilson, W. D.; Hurley, L. H. J. Am. and provide constant mixture by aspiration and dispensing during
Chem. Soc. 2005, 127, 2944–2959. measurements. This experimental arrangement maintains a ho-
(6) (a) Lyon, L. A.; Pena, D. J.; Natan, M. J. J. Phys. Chem. B 1999, 103, 5826– mogeneous solution and reproducible hydrodynamic conditions.
5831. (b) Link, S.; El-Sayad, M. A. J. Phys. Chem. B 1999, 103, 4212–4217.
(c) Decher, G.; Natan, M.; Peschel, S.; Smith, E. A. In Book of Abstracts, For the detailed experiment, the SPR gold film was first immersed
219th ACS National Meeting, San Francisco, American Chemical Society: into the aptamer solution for 12 h in order to assemble the
Washington, DC,2000. monolayer of aptamer. Then the modified gold film was thor-
(7) (a) He, L.; Musick, M. D.; Nicewarner, S. R.; Salinas, F. G.; Benkovic, S. J.;
Natan, M. J.; Keating, C. D. J. Am. Chem. Soc. 2000, 122, 9071–9077. (b) oughly rinsed with 50 mM Tris-HCl buffer and water to remove
Lyon, L. A.; Musick, M. D.; Natan, M. J. Anal. Chem. 1998, 70, 5177– the weakly adsorbed aptamer. Aptamer-modified SPR gold film
5183. (c) Tamada, K.; Nakamura, F.; Ito, M.; Li, X. H.; Baba, A. Plasmonics was immersed in 100 µM 6-mercaptohexanol for 1 h to block the
2007, 2, 185–191.
(8) (a) Zayats, M.; Pogorelova, S. P.; Kharitonov, A. B.; Lioubashevski, E. K.; uncovered gold surface. This gold film was used as a sensor
Willner, I. Chem. Eur. J. 2003, 9, 6108–6114. (b) Matsui, J.; Akamatsu, K.; surface to detect the concentration of adenosine. These detection
Hara, N.; Miyoshi, D.; Nawafune, H.; Tamaki, K.; Sugimoto, N. Anal. Chem. steps are similar to a sandwich assay. After the aptamer was
2005, 77, 4282–4285.
(9) (a) Soh, H.; Tokuda, T.; Watanabe, T.; Mishima, K.; Imato, T.; Masadome, immobilized on SPR gold film, a solution with different concentra-
T.; Asano, Y.; Okutani, S.; Niwa, O.; Brown, S. Talanta 2003, 60, 733– tions of adensine was first added to the SPR cell and reacted for
745. (b) Shankaran, D. R.; Gobi, K. V.; Sakai, T.; Matsumoto, K.; Toko, K.;
Miura, N. Biosens. Bioelectron. 2005, 20, 1750–1756. (c) Kim, S. J.; Gobi, (10) Storhoff, J. J.; Elghanian, R.; Mucic, R. C.; Mirkin, C. A.; Letsinger, R. L.
K. V.; Iwasaka, H.; Tanaka, H.; Miura, N. Biosens. Bioelectron. 2007, 23, J. Am. Chem. Soc. 1998, 120, 1959–1964.
701–707. (d) Kawaguchi, T.; Shankaran, D. R.; Kim, S. J.; Gobi, K. V.; (11) Wang, Y. L.; Wei, H.; Li, B. L.; Ren, W.; Guo, S. J.; Dong, S. J.; Wang, E. K.
Matsumoto, K.; Toko, K.; Miura, N. Talanta 2007, 554–560. Chem. Comm. 2007, 5220–5222.

Analytical Chemistry, Vol. 80, No. 18, September 15, 2008 7175
Figure 1. Schematic representation of the SPR biosensor for the detection of the small molecules.

30 min. After that, SPR cell was washed with buffer and water. formed.13,14 Thus, the amounts of Au nanoparticles-tagged comple-
Last, the Au nanoparticles-tagged complementary ss-DNA was mentary ss-DNA bound to the aptamers immobilized on the SPR
added to the SPR cell and the SPR angle-time curve was surface by DNA hybridization will decrease with the increase of
recorded. The inject rate for all samples was 10 µL/s; the volume the amount of aptamers possessing tertiary structure, which
for all samples dispensed in the SPR cell was 40 µL, and the mixing increases with the increasing concentrations of small molecules.
rate for all samples during the process of experiment was Through detecting DNA hybridization of Au nanoaprticles-tagged
10 µL/s. complementary ss-DNA with aptamers by SPR angle-time curves,
we can deduce a reliable relation between the concentrations of
small molecules and SPR angle shift. Based on this principle, a
RESULTS AND DISCUSSION SPR biosensor for small molecules with high sensitivity and
Detection Principle of SPR Biosensor. The principle of the selectivity is designed.
SPR biosensor for detecting small molecules is shown in Figure SPR Spectroscopy Detects the Concentration of Adenos-
1. Aptamers responding to small molecules are first immobilized ine by the Amplifying Effect of Gold Nanoparticles. We employ
on SPR gold film as sensor surface by gold-sulfur affinity. The antiadenosine aptamer/adenosine as a model system to detect the
immobilized aptamers on the SPR gold surface have an ss-DNA sensing ability of this SPR biosensor for small molecules (adenos-
ine in this case). We choose this well-studied system13 so we can
structure. As ss-DNA, it can hybridize with its complementary ss-
compare the sensitivity of the SPR biosensor we developed with
DNA and form a stable duplex structure. In order to enhance the
other methods. Moreover, it is the first time adenosine was
change of SPR signal to result in the DNA hybridization, we label
detected using the SPR biosensor based on an aptamer technique.
the complementary ss-DNA with Au nanoparticles.12 As a result,
During our experiments, we first detected the molecule density
the angle shift of SPR will be greatly increased when Au
of aptamer immobilized on SPR gold film (See Figure S2 in
nanoparticles-tagged complementary ss-DNA is added to the SPR
Supporting Information.). The molecule density of aptamer is 0.67
reaction cell (case 1 in Figure 1). For case 2 in Figure 1, on the
ng/mm2, which is similar to the value detected by other
other hand, when small molecules are first added to the SPR cell,
methods.13d And then, we utilized SPR angle-time curve to
aptamers immobilized on SPR gold film will react with small
observe the change of SPR signal resulting from the conforma-
molecules. Aptamers with a free coil will fold from the outer 3′
tional change of aptamer triggered by adenosine. The results were
end to the inner 5′ end and form a stable and well-defined tertiary
structure with the introduction of the small molecules. The (13) (a) Liu, J. W.; Lu, Y. Anal. Chem. 2004, 76, 1627–1632. (b) Liu, J. W.; Lu,
concentration of small molecule determines the amount of Y. J. Am. Chem. Soc. 2007, 129, 8634–8643. (c) Zhao, W. A.; Chiuman,
W.; Brook, M. A.; Li, Y. F. ChemBioChem 2007, 8, 727–731. (d) Shen, L.;
aptamers possessing a tertiary structure. When more small
Chen, Z.; Li, Y. H.; Jing, P.; Xie, S. B.; He, S. L.; He, P. L.; Shao, Y. H.
molecules are added, more aptamer with tertiary structure will Chem. Commun. 2007, 2169–2171. (e) Li, B. L.; Du, Y.; Wei, H.; Dong,
be formed. Interestingly, the aptamer will be difficult to hybridize S. J. Chem. Commun. 2007, 3780–3782. (f) Zayats, M.; Huang, Y.; Gill, R.;
Ma, C. A.; Willner, I. J. Am. Chem. Soc. 2006, 128, 13666–13667.
with its complementary ss-DNA after its tertiary structure has (14) (a) Green, L. S.; Jellinek, D.; Jenison, R.; Ostman, A.; Heldin, C. H.; Janjic,
N. Biochemistry 1996, 35, 14413–14424. (b) Wu, Z. M.; Guo, M. M.; Zhang,
S, B.; Chen, C. R.; Jiang, J. H.; Shen, G. L.; Yu, R. Q. Anal. Chem. 2007,
(12) (a) Liu, J. W.; Lu, Y. Nat. Protoc.q 2007, 1, 246–252. 79, 2933–2939.

7176 Analytical Chemistry, Vol. 80, No. 18, September 15, 2008
 DNA. It has been demonstrated that an aptamer can bind tightly
and specifically to a variety of small molecules to form a tertiary
complex with a binding constant greater than that of an ordinary
DNA duplex.14 In other words, the hybridization interaction
between ss-DNA and aptamer will not induce the change of
aptamer from its tertiary structure to a duplex structure. Some
other references also confirm that the presence of a small molecule
will result in a DNA duplex consisting of aptamer and its partial
cDNA changing its conformation to bind its small molecule, with
the short complementary oligonucleic acid being released.13 In
Figure 2. SPR angle-time curves observing the conformational our experiment, we chose a similar short-chain ss-DNA sequence
change of aptamer without adenosine (curve 0) and with 1 mM to tag Au nanoparticles. So Au nanoparticles-tagged complemen-
adenosine (curve 1) in 50 mM Tris-HCl buffer. tary ss-DNA only hybridizes with the rest of the aptamers
possessing a free coil structure. Thus, the decrease of SPR angle
shift can be explained by the reduction of the molecule density
of aptamers with free coil structure on SPR gold film. When the
concentration of adenosine is further increased, we can observe
the SPR angle shift gradually decreases. However, the trend of
decrease will reduce after adenosines with high concentrations
are added, especially when the concentration of adenosine is larger
than 10-6 M. By analyzing the change of SPR angle shift with
the concentrations of adenosine, we obtain a good linear relation-
ship between the logarithms of adenosine concentrations and the
SPR angle shift over a range of 1 × 10-9-1 × 10-6 M (shown in
the inset of Figure 3). The detection results for analyzing the
concentration of adenosine are comparable with other optical and
Figure 3. SPR angle-time curves for the detection of the binding electrochemical aptasensors11 and are much superior to the
process between Au nanoparticles-tagged complementary ss-DNA
detection results obtained by general SPR sensor based on a
and aptamer after the aptamer is reacted with different concentrations
of adenosine for 30 min. Inset: curve describing the linear relationship molecularly imprinted technique.15 It must be pointed out that a
between the SPR angle shift and the adenosine concentrations. nonspecific adsorption can be found in this SPR biosensor. From
Figure 3, we can see that the SPR angle-time curves of Au
shown in Figure 2. The SPR angle-time curve was almost nanoparticles-tagged complementary ss-DNA binding with aptam-
unchanged in the absence of adenosine (curve 0). The SPR angle er immobilized on gold surface still change ∼0.1° even when the
slightly increased with time after 1 mM adenosine was added to aptamer reacts with 1 mM adenosine. The main reason for this
the cell (curve 1). It means adenosine binds with aptamer and phenomenon may come from the stereohindrance effect of the
results in the conformational change of aptamer. However, the aptamer possessing different structures. After adenosine is added
SPR angle would rapidly decrease after we washed SPR gold film to the cell, most of the free-coiled aptamer react with adenosine
with buffer. The angle shift resulting from the binding of and form its tertiary structure. However, the formation of a tertiary
adenosine was ∼0.004°. The value of angle shift is so small that structure of an aptamer may inhibit the further reaction between
we could not detect the trace concentration of adenosine by SPR of its adjacent free-coiled aptamer and adenosine. Thus, the
spectroscopy directly. Thus, it was necessary to amplify the SPR nonspecific adsorption occurs after Au nanoparticles-tagged
signal. We adopted a three-step procedure similar to a sandwich complementary ss-DNA is added.
immunoassay to complete the amplified detection of adenosine. Selectivity of SPR Biosensor for Detection of Adenosine.
The procedure was as follows: First, immobilize aptamer to SPR The selectivity of the sensing system is another important
gold film; second, add the different concentrations of adenosine parameter for a biosensor. An excellent biosensor should not only
to SPR cell; and last, wash SPR cell and add Au nanoparticles- possess a good sensitivity but should also have a good selectivity.
tagged complementary ss-DNA. The SPR angle-time curve is In order to detect the selectivity of the present biosensor, we chose
used to monitor the reaction progress between Au nanoparticles- three kinds of compounds (uridine, cytidine, and guanosine),
tagged complementary ss-DNA and aptamer after different con- which belong to the nucleosides family, and have a structure
centrations of adenosine are reacted with the aptamer immobilized similar to that of adenosine. Figure 4 exhibits different SPR
on SPR gold film for 30 min. The results are shown in Figure 3. angle-time binding curves between Au nanopartocles-tagged
In the absence of adenosine, aptamers immobilized on SPR gold complementary ss-DNA and aptamer after aptamer immobilized
film directly hybridized with Au nanoparticles-tagged complemen- on SPR gold film reacted with all kinds of analytes for 30 min.
tary ss-DNA and resulted in the largest SPR angle shift (∼0.831°). The first curve from top to down in Figure 4 is a baseline, which
After 1 nM adenosine was added to the SPR cell, the SPR angle represents the SPR binding curve of Au nanopartocles-tagged
shift resulting from the binding of Au nanoparticles-tagged complementary ss-DNA with aptamer without any addition. The
complementary ss-DNA decreases (∼0.748°) because parts of
(15) (a) Taniwaki, K.; Hyakutake, A.; Aoki, T.; Yoshikawa, M.; Guiver, M. D.;
aptamers react with adenosine and form a tertiary structure, which Robertson, G. P. Anal. Chim. Acta 2003, 489, 191–198. (b) Yoshikawa,
cannot hybridize with Au nanoparticles-tagged complementary ss- M.; Guiver, M. D.; Robertson, G. P. J. Mol. Struct. 2005, 739, 41–46.

Analytical Chemistry, Vol. 80, No. 18, September 15, 2008 7177
 CONCLUSION
A novel strategy for the construction of a SPR sensing system
for highly sensitive adenosine detection is described. This method
combines the advantage of an aptamer technique with the
amplifying effect of Au nanoparticles to extend the application of
SPR spectroscopy for the detection of small molecules. Using this
method, we have successfully detected adenosine over a range
of 1 × 10-9-1 × 10-6 M by SPR spectroscopy. Meanwhile, we
also confirm that the SPR biosensor based on this method
possesses a good selectivity for small-molecule targets. More
significantly, it is fairly easy to generalize this strategy to detect
Figure 4. SPR angle-time curves for the detection of the binding a spectrum of targets by SPR spectroscopy using different
process between Au nanoparticles-tagged complementary ss-DNA aptamers. Therefore, it is expected that this method may offer a
and aptamer after aptamer is reacted with different kinds of analytes new direction in designing high-performance SPR biosensors for
for 30 min. sensitive and selective detection of a wide spectrum of small
other curves from top to down are the SPR binding curve of Au molecules.
nanopartocles-tagged complementary ss-DNA with aptamer after
the aptamer reacted with 1 mM cytidine, guanosine, uridine, and SUPPORTING INFORMATION AVAILABLE
adenosine for 30 min, respectively. Compared with the baseline, Additional information as noted in text. This material is
we can see the SPR angle shift slightly decreases after the aptamer available free of charge via the Internet at http://pubs.acs.org.
reacted with 1 mM cytidine, guanosine, and uridine. Oppositely,
The SPR angle shift greatly decreases after addition of 1 mM
adenosine. These results confirm that the developed strategy has Received for review June 23, 2008. Accepted July 21,
sufficient specificity and adenosine can be identified with high 2008.
selectivity. AC801281C

7178 Analytical Chemistry, Vol. 80, No. 18, September 15, 2008