Analytical Chemistry Chemical Cytometry Quantitates Superoxide
Analytical Chemistry Chemical Cytometry Quantitates Superoxide
Analytical Chemistry Chemical Cytometry Quantitates Superoxide
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Figure 6. (A) SPR angle-time curves of the separation products obtained after Fe3O4 MNP-antiadenosine aptamer conjugates are reacted<br />
with different concentrations of adenosine for 30 min. Inset: The linear relationship between the logarithms of adenosine concentrations and the<br />
SPR angle shift resulting from the binding of Fe3O4 MNP-antiadenosine aptamer conjugates. (B) SPR angle-time curves of 1 µM antiadenosine<br />
aptamer without adenosine (red line) and after react with 1 mM adenosine for 30 min (black line).<br />
10 nM adenosine because parts of aptamers on Fe3O4 MNPs<br />
react with adenosine and form its tertiary structure, which<br />
cannot hybridize with its complementary ss-DNA. With the<br />
further increase of the concentration of adenosine added to<br />
the Fe3O4 MNP-antiadenosine aptamer conjugate solution,<br />
the SPR angle shift continuously decreases until most of the<br />
antiadenosine aptamer binds with adenosine. By analyzing<br />
the change of SPR angle shift with the concentrations of<br />
adenosine, a good linear relationship between the logarithms<br />
of adenosine concentrations and the SPR angle shift is<br />
obtained with a range of 10-1 × 10 4 nM (shown in the insert<br />
of Figure 6A). This detection result is comparable with most<br />
other aptasensors 44 and is a little lower than that of detection<br />
results from SPR aptasensor which utilize Au NPs as an<br />
amplification reagent 20,29 but much superior to the detection<br />
results obtained by a general SPR sensor based on a molecularly<br />
imprinted technique. 45,46 It should be pointed out that the<br />
SPR angle shift only decreases to ∼107.33 m° even when the<br />
aptamer reacts with 1 mM adenosine. This nonspecific adsorption<br />
may come from the stereohindrance effect of aptamer<br />
possessing different structures. After adenosine is added to<br />
Fe3O4 MNP-aptamer conjugates, most free-coiled aptamers<br />
react with adenosine and form its tertiary structure. However,<br />
the formation of a tertiary structure of an aptamer may<br />
inhibit the further reaction between its adjacent free-coiled<br />
aptamer and adenosine. To clearly show the amplification<br />
effect of Fe3O4 MNPs, as a comparison, the SPR response<br />
resulting from the binding of aptamer without MNP is also<br />
studied. The black line in Figure 6B is the SPR angle shift<br />
resulting from the binding of 1 µM antiadenosine aptamer after<br />
antiadenosine aptamer reactes with 1 mM adenosine for 0.5 h.<br />
Only a 24.6 m° SPR angle shift is observed. Although the SPR<br />
angle shift resulting from the binding of antiadenosine aptamer<br />
could increase to 65.7 m° (Red line in Figure 6B) in the absence<br />
of adenosine, this value is still much lower than that of the<br />
(44) Wang, Y. L.; Wei, H.; Li, B. L.; Ren, W.; Guo, S. J.; Dong, S. J.; Wang, E. K.<br />
Chem. Commun. 2007, 5220–5222.<br />
(45) Taniwaki, K.; Hyakutake, A.; Aoki, T.; Yoshikawa, M.; Guiver, M. D.;<br />
Robertson, G. P. Anal. Chim. Acta 2003, 489, 191–198.<br />
(46) Yoshikawa, M.; Guiver, M. D.; Robertson, G. P. J. Mol. Struct. 2005, 739,<br />
41–46.<br />
6788 <strong>Analytical</strong> <strong>Chemistry</strong>, Vol. 82, No. 16, August 15, 2010<br />
Figure 7. SPR angle-time curves of the separation products<br />
obtained after Fe3O4 MNP-antiadenosine aptamer conjugates are<br />
reacted with different analytes for 30 min.<br />
value resulting from the binding of Fe3O4 MNP-antiadenosine<br />
aptamer conjugates (1082.94 m°). Besides sensitivity, the<br />
specification of aptamer promises the selectivity of the<br />
present SPR sensor for adenosine. Figure 7 exhibits SPR<br />
angle-time curves of the separation products obtained after<br />
Fe3O4 MNP-antiadenosine aptamer conjugates are reacted<br />
with different analytes for 30 min. It could be easily seen<br />
that the addition of Fe3O4 MNP-antiadenosine aptamer<br />
conjugates which are reacted with 1 × 10 6 nM cytidine,<br />
guanosine, and uridine result in big SPR angle shifts.<br />
However, The SPR angle shift only increases a little after<br />
Fe3O4 MNP-antiadenosine aptamer conjugates are reacted<br />
with 1 × 10 6 nM adenosine. These results not only demonstrate<br />
that Fe3O4 MNPs could greatly enhance the SPR signal<br />
but also, more importantly, give us an important indication<br />
that SPR spectroscopy could be an excellent candidate for<br />
detecting an MNP-based separation product.<br />
CONCLUSION<br />
In summary, the SPR response of the carboxyl group<br />
modified Fe3O4 MNPs of two different sizes onto an amino<br />
group modified SPR gold substrate has been studied. The<br />
results show the monolayer adsorption of Fe3O4 MNPs could