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The ability of two-dimensional infrared (2D-IR) spectroscopy to measure the amide I band of proteins in H2O rather than D2O-based solvents by evading the interfering water signals has enabled in vivo studies of proteins under physiological conditions and in biofluids. Future exploitation of 2D-IR in analytical settings, from diagnostics to protein screening, will, however, require comparisons between multiple datasets, necessitating control of data collection protocols to minimize measurement-to-measurement inconsistencies. Inspired by analytical spectroscopy applications in other disciplines, we describe a workflow for pre-processing 2D-IR data that aims to simplify spectral cross-comparisons. Our approach exploits the thermal water signal that is collected simultaneously with, but is temporally separated from the amide I response to guide custom baseline correction and spectral normalization strategies before combining them with Principal Component noise reduction tools. Case studies show that application of elements of the pre-processing workflow to previously published data enables improvements in quantification accuracy and detection limits. We subsequently apply the complete workflow in a new pilot study, testing the ability of a prototype library of 2D-IR spectra to quantify the four major protein constituents of blood serum in a single, label-free measurement. These advances show progress toward the robust data handling strategies that will be necessary for future applications of 2D-IR to pharmaceutical or biomedical problems.
- 2D-IR spectroscopy
- pre-processing steps
- protein library
FingerprintDive into the research topics of 'Measuring proteins in H2O using 2D-IR spectroscopy: pre-processing steps and applications toward a protein library'. Together they form a unique fingerprint.
- 1 Finished
Nordon, A. & Baker, M.
1/06/20 → 30/11/23
Data for: "Measuring Proteins in H2O using 2D-IR Spectroscopy; Pre-Processing Steps and Applications towards a Protein Library"
Rutherford, S. (Creator), Nordon, A. (Creator), Hunt, N. (Creator), Baker, M. (Creator), Greetham, G. (Creator) & Parker, A. (Creator), University of Strathclyde, 1 Nov 2022