Department of Chemical Engineering, Worcester Polytechnic Institute, 100 Institute Road, Worcester, Massachusetts 01609, USA.
Anal Chem. 2010 Aug 15;82(16):6782-9. doi: 10.1021/ac100812c.
Magnetic nanoparticles (MNPs) have been frequently used in bioseparation, but their applicability in bioassays is limited due to their extremely small size so that sensitive detection is difficult to achieve using a general technique. Here, we present an amplification technique using MNPs for an enhanced surface plasmon resonance (SPR) bioassay. The amplification effect of carboxyl group modified Fe(3)O(4) MNPs of two sizes on SPR spectroscopy is first demonstrated by assembling MNPs on amino group modified SPR gold substrate. To further evaluate the feasibility of the use of Fe(3)O(4) MNPs in enhancing a SPR bioassay, a novel SPR sensor based on an indirect competitive inhibition assay (ICIA) is developed for detecting adenosine by employing Fe(3)O(4) MNP-antiadenosine aptamer conjugates as the amplification reagent. The results confirm that Fe(3)O(4) MNPs can be used as a powerful amplification agent to provide a sensitive approach to detect adenosine by SPR within the range of 10-10,000 nM, which is much superior to the detection result obtained by a general SPR sensor. Importantly, the present detection methodology could be easily extended to detect other biomolecules of interest by changing the corresponding aptamer in Fe(3)O(4) MNP-aptamer conjugates. This novel technique not only explores the possibility of the use of SPR spectroscopy in a highly sensitive detection of an MNP-based separation product but also offers a new direction in the use of Fe(3)O(4) MNPs as an amplification agent to design high performance SPR biosensors.
磁性纳米粒子(MNPs)已被广泛应用于生物分离,但由于其极小的尺寸,其在生物测定中的适用性受到限制,因此使用一般技术很难实现灵敏检测。在这里,我们提出了一种使用 MNPs 的放大技术,用于增强表面等离子体共振(SPR)生物测定。首先通过将 MNPs 组装到氨基修饰的 SPR 金基底上来证明两种尺寸的羧基修饰的 Fe(3)O(4) MNPs 对 SPR 光谱的放大效果。为了进一步评估 Fe(3)O(4) MNPs 在增强 SPR 生物测定中的可行性,我们开发了一种基于间接竞争抑制测定(ICIA)的新型 SPR 传感器,用于通过使用 Fe(3)O(4) MNP-抗腺苷适配体缀合物作为放大试剂来检测腺苷。结果证实,Fe(3)O(4) MNPs 可用作强大的放大剂,通过 SPR 在 10-10000 nM 范围内提供灵敏的腺苷检测方法,这比普通 SPR 传感器的检测结果要好得多。重要的是,通过改变 Fe(3)O(4) MNP-适配体缀合物中的相应适配体,本检测方法可以很容易地扩展到检测其他感兴趣的生物分子。这项新技术不仅探索了 SPR 光谱在基于 MNPs 的分离产物的高灵敏度检测中的应用可能性,而且还为将 Fe(3)O(4) MNPs 用作放大剂设计高性能 SPR 生物传感器提供了新的方向。