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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.

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


Authors

Senthooran Selvarajah

Mohamed R. Hamed

Carolyn Tubby

Ian Todd



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., Harrison, T., & 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
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/
Contract Date Feb 23, 2017

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