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

Selvarajah, Senthooran; Negm, Ola H.; Hamed, Mohamed R.; Tubby, Carolyn; Todd, Ian; Tighe, Patrick J.; Harrison, Tim; Fairclough, Lucy C.

Abstract

© 2014 Senthooran Selvarajah et al. 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.

Citation

Selvarajah, S., Negm, O. H., Hamed, M. R., Tubby, C., Todd, I., Tighe, P. J., …Fairclough, L. C. (2014). Development and validation of protein microarray technology for simultaneous inflammatory mediator detection in human sera. Mediators of Inflammation, 2014, 1-12. https://doi.org/10.1155/2014/820304

• Journal Article Type: Article
• Acceptance Date: Sep 8, 2014
• Publication Date: Oct 14, 2014
• Deposit Date: Feb 23, 2017
• Publicly Available Date: Feb 23, 2017
• Journal: Mediators of Inflammation
• Print ISSN: 0962-9351
• Electronic ISSN: 1466-1861
• Publisher: Hindawi Publishing Corporation
• Peer Reviewed: Peer Reviewed
• Volume: 2014
• Article Number: 820304
• Pages: 1-12
• DOI: https://doi.org/10.1155/2014/820304
• Public URL: https://nottingham-repository.worktribe.com/output/738310
• Publisher URL: https://www.hindawi.com/journals/mi/2014/820304/