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International comparability in spectroscopic measurements of protein structure by circular dichroism: CCQM-P59.1

International comparability in spectroscopic measurements of protein structure by circular dichroism: CCQM-P59.1
International comparability in spectroscopic measurements of protein structure by circular dichroism: CCQM-P59.1
Circular dichroism (CD) is a spectroscopic technique that is widely used to obtain information about protein structure, and hence is an important tool with many applications, including the characterization of biopharmaceuticals. A previous inter-laboratory study, CCQM-P59, showed that there was a poor level of comparability between laboratories in CD spectroscopy. In a follow-up study reported here, we achieved our goal of demonstrating improved comparability and data quality, primarily by addressing the problems identified in the previous study, which included cell path-length measurement, instrument calibration and good practice in general. Multivariate analysis techniques (principal component analysis and soft independent modelling of class analogies) were shown to be useful in comparing large spectral data sets and in classifying spectra. However, our results also show that there is more work to be done to improve confidence in the technique as the discrepancies observed were partially due to systematic effects, which the statistical approaches do not consider. We therefore conclude that there is a need for an improved understanding of the uncertainties in CD measurement.
0026-1394
Ravi, Jascindra
d0c93c4d-3a20-4b47-83de-588adc502509
Rakowska, Paulina D
73bf2145-e2fa-46ee-9ad9-f74c80d5a369
Garfagnini, Tommaso
66eda1bb-4217-447a-a136-38ea9edd51a6
Baron, Bruno
b50e1c0a-0f59-4d14-b260-33271c3549f9
Charlet, Philippe
6fd8b569-7fa1-4aad-bbc7-d4e09b3aa639
Milev, Stoyan
a19a3dec-581c-4867-96da-5ce24d20b7ef
Lorenz, Julie DeSa
cf4cd11f-bcaf-4aa5-b2e4-dce72f40309c
Plusquellic, David
fea845f2-5e4a-4efb-a6c6-f92befb99f77
Wien, Frank
95f2d0e0-b9bc-40b1-b144-289f07ffe565
Ravi, Jascindra
d0c93c4d-3a20-4b47-83de-588adc502509
Rakowska, Paulina D
73bf2145-e2fa-46ee-9ad9-f74c80d5a369
Garfagnini, Tommaso
66eda1bb-4217-447a-a136-38ea9edd51a6
Baron, Bruno
b50e1c0a-0f59-4d14-b260-33271c3549f9
Charlet, Philippe
6fd8b569-7fa1-4aad-bbc7-d4e09b3aa639
Milev, Stoyan
a19a3dec-581c-4867-96da-5ce24d20b7ef
Lorenz, Julie DeSa
cf4cd11f-bcaf-4aa5-b2e4-dce72f40309c
Plusquellic, David
fea845f2-5e4a-4efb-a6c6-f92befb99f77
Wien, Frank
95f2d0e0-b9bc-40b1-b144-289f07ffe565

Ravi, Jascindra, Rakowska, Paulina D, Garfagnini, Tommaso, Baron, Bruno, Charlet, Philippe, Milev, Stoyan, Lorenz, Julie DeSa, Plusquellic, David and Wien, Frank (2010) International comparability in spectroscopic measurements of protein structure by circular dichroism: CCQM-P59.1. Metrologia, 47 (6). (doi:10.1088/0026-1394/47/6/001).

Record type: Article

Abstract

Circular dichroism (CD) is a spectroscopic technique that is widely used to obtain information about protein structure, and hence is an important tool with many applications, including the characterization of biopharmaceuticals. A previous inter-laboratory study, CCQM-P59, showed that there was a poor level of comparability between laboratories in CD spectroscopy. In a follow-up study reported here, we achieved our goal of demonstrating improved comparability and data quality, primarily by addressing the problems identified in the previous study, which included cell path-length measurement, instrument calibration and good practice in general. Multivariate analysis techniques (principal component analysis and soft independent modelling of class analogies) were shown to be useful in comparing large spectral data sets and in classifying spectra. However, our results also show that there is more work to be done to improve confidence in the technique as the discrepancies observed were partially due to systematic effects, which the statistical approaches do not consider. We therefore conclude that there is a need for an improved understanding of the uncertainties in CD measurement.

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Published date: 4 October 2010

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Local EPrints ID: 445180
URI: http://eprints.soton.ac.uk/id/eprint/445180
ISSN: 0026-1394
PURE UUID: 7f4c7530-2106-4477-a448-f7434e60e149
ORCID for Paulina D Rakowska: ORCID iD orcid.org/0000-0002-3710-8395

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Date deposited: 24 Nov 2020 17:34
Last modified: 28 Apr 2022 02:31

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Contributors

Author: Jascindra Ravi
Author: Tommaso Garfagnini
Author: Bruno Baron
Author: Philippe Charlet
Author: Stoyan Milev
Author: Julie DeSa Lorenz
Author: David Plusquellic
Author: Frank Wien

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