Influence of liquid medium and surface morphology on the response of QCM during immobilization and hybridization of short oligonucleotides

被引:30
作者
Ha, TH
Kim, S
Lim, G
Kim, K [1 ]
机构
[1] Seoul Natl Univ, Sch Chem & Mol Engn, Lab Intelligent Interfaces, Seoul 151742, South Korea
[2] Samsung Adv Inst Technol, Biochip Project Team, Suwon 440600, South Korea
关键词
quartz crystal microbalance; oligonucleotides; single-stranded DNA;
D O I
10.1016/j.bios.2004.02.027
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
With the goal of developing a quartz crystal microbalance (QCM)-based DNA sensor, we have conducted an in situ QCM study along with fluorescence measurements using oligonucleotides (15-mer) as a model single-stranded DNA (ss-DNA) in two different aqueous buffer solutions; the sequence of 15-mer is a part of iduronate-2-sulphate exon whose mutation is known to cause Hunter syndrome, and the 15-mer is thiolated to be immobilized on the Au-coated quartz substrate. The fluorescence data indicate that the initial immobilization as well as the subsequent hybridization with a complementary strand is hardly dependent on the kind of buffer solution. In contrast, the mass increases deducible from the decrease of QCM frequency via the Sauerbrey equation are 2.7-6.2 and 3.0-4.4 times larger than the actual mass increases, as reflected in the fluorescence measurements, for the immobilization and the subsequent hybridization processes, respectively. Such an overestimation is attributed to the trapping of solvent as well as the formation of quite a rigid hydration layer associated with the higher viscosities and/or densities of the buffer solutions. Another noteworthy observation is the excessively large frequency change that occurs when the gold electrode is deposited in advance with Au nanoparticles. This clearly illustrates that the QCM detection of DNA hybridization is also affected greatly by the surface morphology of the electrode. These enlarged signals are altogether presumed to be advantageous when using a QCM system as an in situ probing device in DNA sensors. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:378 / 389
页数:12
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