Urinary 8-oxo-7,8-dihydroguanosine values determined by a modified ELISA improves agreement with HPLC-MS/MS

0398237 - ÚEM 2014 RIV US eng J - Journal Article
Rössner ml., Pavel - Mistry, V. - Singh, R. - Šrám, Radim - Cooke, M. S.
Urinary 8-oxo-7,8-dihydroguanosine values determined by a modified ELISA improves agreement with HPLC-MS/MS.
Biochemical and Biophysical Research Communications. Roč. 440, č. 4 (2013), s. 725-730. ISSN 0006-291X. E-ISSN 1090-2104
R&D Projects: GA ČR GAP503/11/0084; GA ČR GA13-13458S
Institutional support: RVO:68378041
Keywords : 8-oxodG * urine * ELISA
Subject RIV: DN - Health Impact of the Environment Quality
Impact factor: 2.281, year: 2013

ELISA is widely used for urinary 8-oxo-7.8-dihydro-2´-deoxyguanosine (8-oxodG) analysis. However, a lack of agreement between ELISA and chromatographic methods confines its application to the assessment of relative urinary 8-oxodG level. We investigated various ELISA modifications, seeking optimal conditions that would yield a good agreement between ELISA and HPLC-MS/MS. It was found, that purification of urine by solid phase extraction, then incubation with the anti-8-oxodG antibody at 40 C overnight and subsequent normalization of 8-oxodG levels per urinary creatinine resulted in a near-perfect correlation and agreement in mean levels between ELISA and HPLC-MS/MS. Our data show that, after introduction of a simple modification, ELISA quantification urinary 8-oxodG substantially improves. This method represents a significant advance for the ELISA, and encouraging its use in more studies adding to our knowledge of the role of this biomarker of oxidative stress in health and disease.
Permanent Link: http://hdl.handle.net/11104/0225748