Essential modification of the Sircol Collagen Assay for the accurate quantification of collagen content in complex protein solutions

Lareu, Ricky R.; Zeugolis, Dimitrios I.; Abu-Rub, Mohammad; Pandit, Abhay; Raghunath, Michael

doi:10.1016/j.actbio.2010.02.004

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
Title:	Essential modification of the Sircol Collagen Assay for the accurate quantification of collagen content in complex protein solutions
Authors:	Lareu, Ricky R. Zeugolis, Dimitrios I. Abu-Rub, Mohammad Pandit, Abhay Raghunath, Michael
DOI:	10.1016/j.actbio.2010.02.004
Published in:	Acta Biomaterialia
Volume(Issue):	6
Issue:	8
Page(s):	3146
Pages to:	3151
Issue Date:	2010
Publisher / Ed. Institution:	Elsevier
ISSN:	1742-7061 1878-7568
Language:	English
Subject (DDC):	572: Biochemistry
Abstract:	Collagen contains the unique imino acid hydroxyproline (HyPro), which is involved in the stabilization of this triple helical molecule. The concentration of HyPro is customarily used to calculate the total collagen content in a cell culture environment and in acid hydrolysates of normal and pathophysiological tissues. Radiolabelling, chromatographic and calorimetric assays have been developed over the years for the accurate determination of collagen content through HyPro estimation. Recently, the Sircol Collagen Assay (SCA) has been almost exclusively adopted as the fastest and simplest colorimetric method for the determination of collagen concentration in complex protein solutions. We show here that the colorimetric SCA, which is based on the binding of Sirius red (SR) to collagen, is flawed by interference of non-collagenous proteins (e.g. serum). In fact, we demonstrate that SCA in cell culture systems and tissue hydrolysates results in a dramatic overestimation of collagen content ranging from 3- to 24-fold. In order to rescue this otherwise very practical assay, we introduce a simple purification procedure that allows the removal of interfering non-collagenous proteins from culture media and tissue samples so that accurate measurements with SCA are now possible.
URI:	https://digitalcollection.zhaw.ch/handle/11475/12209
Fulltext version:	Published version
License (according to publishing contract):	Licence according to publishing contract
Departement:	Life Sciences and Facility Management
Organisational Unit:	Institute of Chemistry and Biotechnology (ICBT)
Appears in collections:	Publikationen Life Sciences und Facility Management

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Lareu, R. R., Zeugolis, D. I., Abu-Rub, M., Pandit, A., & Raghunath, M. (2010). Essential modification of the Sircol Collagen Assay for the accurate quantification of collagen content in complex protein solutions. Acta Biomaterialia, 6(8), 3146–3151. https://doi.org/10.1016/j.actbio.2010.02.004

Lareu, R.R. et al. (2010) ‘Essential modification of the Sircol Collagen Assay for the accurate quantification of collagen content in complex protein solutions’, Acta Biomaterialia, 6(8), pp. 3146–3151. Available at: https://doi.org/10.1016/j.actbio.2010.02.004.

R. R. Lareu, D. I. Zeugolis, M. Abu-Rub, A. Pandit, and M. Raghunath, “Essential modification of the Sircol Collagen Assay for the accurate quantification of collagen content in complex protein solutions,” Acta Biomaterialia, vol. 6, no. 8, pp. 3146–3151, 2010, doi: 10.1016/j.actbio.2010.02.004.

LAREU, Ricky R., Dimitrios I. ZEUGOLIS, Mohammad ABU-RUB, Abhay PANDIT und Michael RAGHUNATH, 2010. Essential modification of the Sircol Collagen Assay for the accurate quantification of collagen content in complex protein solutions. Acta Biomaterialia. 2010. Bd. 6, Nr. 8, S. 3146–3151. DOI 10.1016/j.actbio.2010.02.004

Lareu, Ricky R., Dimitrios I. Zeugolis, Mohammad Abu-Rub, Abhay Pandit, and Michael Raghunath. 2010. “Essential Modification of the Sircol Collagen Assay for the Accurate Quantification of Collagen Content in Complex Protein Solutions.” Acta Biomaterialia 6 (8): 3146–51. https://doi.org/10.1016/j.actbio.2010.02.004.

Lareu, Ricky R., et al. “Essential Modification of the Sircol Collagen Assay for the Accurate Quantification of Collagen Content in Complex Protein Solutions.” Acta Biomaterialia, vol. 6, no. 8, 2010, pp. 3146–51, https://doi.org/10.1016/j.actbio.2010.02.004.