Dual-Platform Proteomics Study of Plasma Biomarkers in Pediatric Patients Undergoing Cardiopulmonary Bypass

Department of Pediatrics, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, USA.
Pediatric Research (Impact Factor: 2.31). 03/2010; 67(6):641-9. DOI: 10.1203/PDR.0b013e3181dceef5
Source: PubMed


Plasma samples from pediatric cardiac patients undergoing cardiopulmonary bypass (CPB) procedures were used to identify and characterize patterns of changes in potential biomarkers related to tissue damage and inflammation. These included proteins associated with systemic inflammatory response syndrome. Potential biomarkers were identified using a dual-platform proteomics approach requiring approximately 150 microL of plasma, which included two-dimensional difference gel electrophoresis (2D-DIGE) and a multiplexed immunoassay. Methods used in the dual approach measured levels of 129 proteins in plasma from pediatric CPB patients. Of these, 70 proteins changed significantly (p<0.05) between time points, and 36 of these retained significance after the highly stringent Bonferroni correction [p<0.001 for 2D-DIGE and p<0.00056 for multianalyte profile (MAP) assays]. Many of the changing proteins were associated with tissue damage, inflammation, and oxidative stress. This study uses a novel approach that combines two discovery proteomics techniques to identify a pattern of potential biomarkers changing after CPB. This approach required only 150 microL of plasma per time point and provided quantitative information on 129 proteins. The changes in levels of expression of these proteins may provide insight into the understanding, treatment, and prevention of systemic inflammation, thereby helping to improve the outcomes of pediatric CPB patients.

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Available from: John L Myers, Apr 15, 2014
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    • "These procedures have been described in detail previously [29,30,123]. Information about the 2D-DIGE study is provided in a form that complies with the most recent version <> of Minimum Information About a Proteomics Experiment – Gel Electrophoresis (MIAPE-GE) standards currently under development by the Human Proteome Organization Proteomics Standards Initiative (see Additional file 6). "
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    • "We have used this procedure in previous studies for other types of protein samples [26,46,143] but a detailed account, including many modifications and refinements appears below. For identification of spots, all 791 protein spots from the experiment were picked from the picking gel using a robot-directed spot picker (Ettan Spot Picker, GE Healthcare). "
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