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Artículo

Beyond Fourier Transform Infrared Spectroscopy: External Cavity Quantum Cascade Laser-Based Mid-infrared Transmission Spectroscopy of Proteins in the Amide i and Amide II Region

Schwaighofer, Andreas; Montemurro, MilagrosIcon ; Freitag, Stephan; Kristament, Christian; Culzoni, Maria JuliaIcon ; Lendl, Bernhard
Fecha de publicación: 06/2018
Editorial: American Chemical Society
Revista: Analytical Chemistry
ISSN: 0003-2700
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Química Analítica

Resumen

In this work, we present a setup for mid-IR measurements of the protein amide I and amide II bands in aqueous solution. Employing a latest generation external cavity-quantum cascade laser (EC-QCL) at room temperature in pulsed operation mode allowed implementing a high optical path length of 31 μm that ensures robust sample handling. By application of a data processing routine, which removes occasionally deviating EC-QCL scans, the noise level could be lowered by a factor of 4. The thereby accomplished signal-to-noise ratio is better by a factor of approximately 2 compared to research-grade Fourier transform infrared (FT-IR) spectrometers at equal acquisition times. Employing this setup, characteristic spectral features of three representative proteins with different secondary structures could be measured at concentrations as low as 1 mg mL -1 . Mathematical evaluation of the spectral overlap confirms excellent agreement of the quantum cascade laser infrared spectroscropy (QCL-IR) transmission measurements with protein spectra acquired by FT-IR spectroscopy. The presented setup combines performance surpassing FT-IR spectroscopy with large applicable optical paths and coverage of the relevant spectral range for protein analysis. This holds high potential for future EC-QCL-based protein studies, including the investigation of dynamic secondary structure changes and chemometrics-based protein quantification in complex matrices.
Palabras clave: FT-IR SPECTROSCOPY , EC-QCL , PROTEINS
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/85452
DOI: http://dx.doi.org/10.1021/acs.analchem.8b01632
URL: https://pubs.acs.org/doi/10.1021/acs.analchem.8b01632
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Articulos(CCT - SANTA FE)
Articulos de CTRO.CIENTIFICO TECNOL.CONICET - SANTA FE
Citación
Schwaighofer, Andreas; Montemurro, Milagros; Freitag, Stephan; Kristament, Christian; Culzoni, Maria Julia; et al.; Beyond Fourier Transform Infrared Spectroscopy: External Cavity Quantum Cascade Laser-Based Mid-infrared Transmission Spectroscopy of Proteins in the Amide i and Amide II Region; American Chemical Society; Analytical Chemistry; 90; 11; 6-2018; 7072-7079
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