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A label-free sensing method for phosphopeptides using two-layer gold nanoparticle-based localized surface plasma resonance spectroscopy

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Abstract

In this study, a new type of localized surface plasmon resonance (LSPR) sensing substrate for phosphopeptides was explored. It has been known that LSPR response for target species is larger in the near-infrared region (NIR) than in the visible region of the electromagnetic spectrum. Several types of noble metal nanoparticles (NPs) with NIR absorption capacities have been previously demonstrated as effective LSPR-sensing nanoprobes. Herein, we demonstrate a straightforward approach with improved sensitivity by simply using layer-by-layer (LBL) spherical Au NPs self-assembled on glass slides as the LSPR-sensing substrates that are responsive in the NIR region of the electromagnetic spectrum. The modified glass slide acquired an LSPR absorption band in the NIR, which resulted from the dipole–dipole interactions between Au NPs. To enable the chip to sense phosphopeptides, the surface of the glass chip was spin-coated with thin titania film (TiO2-Glass@Au NPs). Absorption spectrophotometry was employed as a detection tool. Tryptic digest of α-casein was used as a model sample. The feasibility of using the new LSPR approach for detecting a potential risk factor leading to cancers (i.e., phosphorylated fibrinopeptide A) directly from human serum samples was demonstrated. Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was used to confirm the results.

A label free LSPR sensing of phosphopeptides using the titania coated LBL spherical Au NPs coated glass slide (TiO2-Glass@Au NPs) as the sensing chip is demonstrated in this work

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Acknowledgments

We thank the National Science Council of Taiwan for supporting this work financially. We also thank Dr. Wei-Yu Chen and Ms. Tsai-Jung Yu for their assistance in obtaining the SEM images, and Mr. Kai-Wei Han for his assistance in preparing the figures.

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Authors and Affiliations

  1. Department of Applied Chemistry, National Chiao Tung University, Hsinchu, 300, Taiwan

    Jen-Yi Chen & Yu-Chie Chen

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  1. Jen-Yi Chen
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  2. Yu-Chie Chen
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Correspondence to Yu-Chie Chen.

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Chen, JY., Chen, YC. A label-free sensing method for phosphopeptides using two-layer gold nanoparticle-based localized surface plasma resonance spectroscopy. Anal Bioanal Chem 399, 1173–1180 (2011). https://doi.org/10.1007/s00216-010-4397-x

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  • DOI: https://doi.org/10.1007/s00216-010-4397-x

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