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

Quasnichka, H, Tarlton, JF, Anderson-Mackenzie, JM, Billingham, ME, Bailey, AJ and Pickford, AR (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.

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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
Subjects : Veterinary Medicine
Divisions : Surrey research (other units)
Authors :
Tarlton, JF
Anderson-Mackenzie, JM
Billingham, ME
Bailey, AJ
Pickford, AR
Date : February 2005
DOI : 10.1016/j.jim.2004.12.016
Copyright Disclaimer : © 2005 Elsevier B.V. All rights reserved.
Uncontrolled Keywords : Collagen type I synthesis, Guinea pig, PICP, Competition ELISA, SPR
Depositing User : Symplectic Elements
Date Deposited : 17 May 2017 10:47
Last Modified : 24 Jan 2020 20:03

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