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Uncertainty in measurement of protein circular dichroism spectra

Uncertainty in measurement of protein circular dichroism spectra
Uncertainty in measurement of protein circular dichroism spectra

Circular dichroism (CD) spectroscopy of proteins is widely used to measure protein secondary structure, and to detect changes in secondary and higher orders of structure, for applications in research and in the quality control of protein products such as biopharmaceuticals. However, objective comparison of spectra is challenging because of a limited quantitative understanding of the sources of error in the measurement. Statistical methods can be used for comparisons, but do not provide a mechanism for dealing with systematic, as well as random, errors. Here we present a measurement model for CD spectroscopy of proteins, incorporating the principal sources of uncertainty, and use the model in conjunction with experimental data to derive an uncertainty budget. We show how this approach could be used in practice for the objective comparison of spectra, and discuss the benefits and limitations of this strategy.

biopharmaceutical, circular dichroism, protein structure, spectroscopy
0026-1394
67-79
Cox, Maurice G.
d913ff40-a486-411a-9f0f-48cfcc8ec5e1
Ravi, Jascindra
d0c93c4d-3a20-4b47-83de-588adc502509
Rakowska, Paulina D.
73bf2145-e2fa-46ee-9ad9-f74c80d5a369
Knight, Alex E.
504f9ce0-8a2d-432b-b6ca-08a6b23f335d
Cox, Maurice G.
d913ff40-a486-411a-9f0f-48cfcc8ec5e1
Ravi, Jascindra
d0c93c4d-3a20-4b47-83de-588adc502509
Rakowska, Paulina D.
73bf2145-e2fa-46ee-9ad9-f74c80d5a369
Knight, Alex E.
504f9ce0-8a2d-432b-b6ca-08a6b23f335d

Cox, Maurice G., Ravi, Jascindra, Rakowska, Paulina D. and Knight, Alex E. (2014) Uncertainty in measurement of protein circular dichroism spectra. Metrologia, 51 (1), 67-79. (doi:10.1088/0026-1394/51/1/67).

Record type: Article

Abstract

Circular dichroism (CD) spectroscopy of proteins is widely used to measure protein secondary structure, and to detect changes in secondary and higher orders of structure, for applications in research and in the quality control of protein products such as biopharmaceuticals. However, objective comparison of spectra is challenging because of a limited quantitative understanding of the sources of error in the measurement. Statistical methods can be used for comparisons, but do not provide a mechanism for dealing with systematic, as well as random, errors. Here we present a measurement model for CD spectroscopy of proteins, incorporating the principal sources of uncertainty, and use the model in conjunction with experimental data to derive an uncertainty budget. We show how this approach could be used in practice for the objective comparison of spectra, and discuss the benefits and limitations of this strategy.

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More information

Published date: February 2014
Keywords: biopharmaceutical, circular dichroism, protein structure, spectroscopy

Identifiers

Local EPrints ID: 485935
URI: http://eprints.soton.ac.uk/id/eprint/485935
ISSN: 0026-1394
PURE UUID: a02257d3-9dd9-48b5-b1c9-122d29df6093
ORCID for Paulina D. Rakowska: ORCID iD orcid.org/0000-0002-3710-8395

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Date deposited: 04 Jan 2024 04:56
Last modified: 18 Mar 2024 03:59

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Contributors

Author: Maurice G. Cox
Author: Jascindra Ravi
Author: Alex E. Knight

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