Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera

Selvarajah, Senthooran and Negm, Ola H. and Hamed, Mohamed R. and Tubby, Carolyn and Todd, Ian and Tighe, Patrick J. and Harrison, Tim and Fairclough, Lucy C. (2014) Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera. Mediators of Inflammation, 2014 . 820304/1-820304/12. ISSN 1466-1861

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Abstract

Biomarkers, including cytokines, can help in the diagnosis, prognosis, and prediction of treatment response across a wide range of disease settings. Consequently, the recent emergence of protein microarray technology, which is able to quantify a range of inflammatory mediators in a large number of samples simultaneously, has become highly desirable. However, the cost of commercial systems remains somewhat prohibitive. Here we show the development, validation, and implementation of an in-house microarray platform which enables the simultaneous quantitative analysis of multiple protein biomarkers. The accuracy and precision of the in-house microarray system were investigated according to the Food and Drug Administration (FDA) guidelines for pharmacokinetic assay validation. The assay fell within these limits for all but the very low-abundant cytokines, such as interleukin- (IL-) 10. Additionally, there were no significant differences between cytokine detection using our microarray system and the “gold standard” ELISA format. Crucially, future biomarker detection need not be limited to the 16 cytokines shown here but could be expanded as required. In conclusion, we detail a bespoke protein microarray system, utilizing well-validated ELISA reagents, that allows accurate, precise, and reproducible multiplexed biomarker quantification, comparable with commercial ELISA, and allowing customization beyond that of similar commercial microarrays.

Item Type: Article
Schools/Departments: University of Nottingham, UK > Faculty of Medicine and Health Sciences > School of Life Sciences
Identification Number: https://doi.org/10.1155/2014/820304
Depositing User: Fairclough, Lucy
Date Deposited: 23 Feb 2017 08:38
Last Modified: 23 Feb 2017 16:18
URI: http://eprints.nottingham.ac.uk/id/eprint/40717

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