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
Published in : Acta biomaterialia
Volume(Issue) : 6
Issue : 8
Pages : 3146
Pages to: 3151
Publisher / Ed. Institution : Elsevier
Issue Date: 2010
License (according to publishing contract) : Licence according to publishing contract
Type of review: Peer review (Publication)
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.
Departement: Life Sciences and Facility Management
Organisational Unit: Institute of Chemistry and Biotechnology (ICBT)
Publication type: Article in scientific Journal
DOI : 10.1016/j.actbio.2010.02.004
ISSN: 1742-7061
1878-7568
URI: https://digitalcollection.zhaw.ch/handle/11475/12209
Appears in Collections:Publikationen Life Sciences und Facility Management

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