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Towards label-free mid-infrared protein assays: in-situ formation of bare gold nanoparticles for surface enhanced infrared absorption spectroscopy of bovine serum albumin

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Abstract

Surface enhanced infrared absorption spectroscopy (SEIRAS) with attenuated total reflection (ATR) configuration has been used to determine bovine serum albumin (BSA) protein adsorbed onto bare gold nanoparticles (AuNPs). The AuNPs forming the SEIRA-active substrate were directly synthesized inside the ATR liquid cell taking advantage of stainless steel assisted synthesis via the walls composing the ATR compartment. The formation of the AuNPs can be directly monitored via the enhancement of the corresponding water absorption features. The absorbance of BSA at the AuNPs deposited onto the Si ATR waveguide is significantly enhanced when compared to bare Si, thereby improving the sensitivity of detection. Apparently, the presence of the AuNPs layer at the ATR waveguide surface at the measured conditions does not affect the secondary structure of the protein. Measurements were performed in water and confirmed in deuterium oxide, which significantly reduces the strongly absorbing mid-infrared background of water in the spectral regime of interest. The limits of detection achieved for BSA protein analysis in water and deuterium oxide media are 1.93 and 4.15 mg·L−1, respectively, and the precisions at a 250 mg·L−1 concentration are 11.9% and 8.1%, respectively.

Bovine serum albumin has been determined via mid-infrared surface enhanced infrared absorption spectroscopy using an attenuated total reflection configuration. The assay is taking advantage of the signal enhancement caused by gold nanoparticles that were synthesized in situ by a stainless steel mediated method.

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Acknowledgements

A.I. López-Lorente wishes to thank the Alexander von Humboldt Foundation for the award of a Postdoctoral Fellowship at the Institute of Analytical and Bioanalytical Chemistry (University of Ulm, Germany). This work was in part supported by the Boehringer Ingelheim Ulm University BioCenter (BIU), and by the International Graduate School in Molecular Medicine at the University of Ulm, which provided a graduate student fellowship. Funding by the Horizon 2020 Framework Program of the European Union within the MSCA RISE Project TROPSENSE is also acknowledged.

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  1. Institute of Analytical and Bioanalytical Chemistry, Ulm University, 89081, Ulm, Germany

    Ángela Inmaculada López-Lorente, Pei Wang & Boris Mizaikoff

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  1. Ángela Inmaculada López-Lorente
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  2. Pei Wang
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Correspondence to Ángela Inmaculada López-Lorente.

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López-Lorente, Á.I., Wang, P. & Mizaikoff, B. Towards label-free mid-infrared protein assays: in-situ formation of bare gold nanoparticles for surface enhanced infrared absorption spectroscopy of bovine serum albumin. Microchim Acta 184, 453–462 (2017). https://doi.org/10.1007/s00604-016-2031-0

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  • DOI: https://doi.org/10.1007/s00604-016-2031-0

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