Development of an assay for the quantification of type I collagen synthesis in the guinea pig

Quasnichka, Helen L., Tarlton, John F., Anderson-MacKenzie, Janet M., Billingham, Michael E.J., Bailey, Allen J. and Pickford, Andrew R. 2005. Development of an assay for the quantification of type I collagen synthesis in the guinea pig. Journal of Immunological Methods 297 (1-2) , pp. 133-141. 10.1016/j.jim.2004.12.016

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Abstract

There is a need for a reliable assay for the quantification of collagen type I synthesis in the guinea pig, an important model for many connective tissue diseases. Procollagen type I C-terminal propeptide (PICP) is the established marker of type I collagen synthesis but, to date, no assay has been developed to measure PICP in guinea pig tissue extracts. A monoclonal antibody, known to cross-react with intact guinea pig procollagen type I (anti-PICP), was tested for its ability to bind soluble guinea pig PICP in crude skin extracts using a biosensor. Anti-PICP was immobilised to the surface of a sensor chip and antibody-antigen binding was detected using the phenomenon of surface plasmon resonance (SPR). The binding component in the SPR-immunoassay was identified as PICP by purification and N-terminal sequencing. Guinea pig PICP was purified from skin by gel filtration, ion exchange chromatography and lectin affinity chromatography. Purified PICP was then biotinylated and used with anti-PICP to develop a competition ELISA that was able to selectively and sensitively measure PICP in extracts of guinea pig connective tissue.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Publisher:	Elsevier
ISSN:	0022-1759
Date of Acceptance:	4 December 2004
Last Modified:	11 Mar 2023 02:06
URI:	https://orca.cardiff.ac.uk/id/eprint/134981

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