Development of an assay for the quantification of type I collagen synthesis in the guinea pig
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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.
Full text not available from this repository.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 | |||||||||||||||||||||
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| Subjects : | Veterinary Medicine | |||||||||||||||||||||
| Divisions : | Surrey research (other units) | |||||||||||||||||||||
| Authors : |
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| 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 | |||||||||||||||||||||
| URI: | http://epubs.surrey.ac.uk/id/eprint/829235 |
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