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Open Access Research

Clinical application for the preservation of phospho-proteins through in-situ tissue stabilization

C Bart Rountree12*, Colleen A Van Kirk2, Hanning You12, Wei Ding12, Hien Dang12, Heather D VanGuilder2 and Willard M Freeman2

Author Affiliations

1 Department of Pediatrics, Penn State Hershey Medical Center, 500 University Drive, Hershey, PA 17033

2 Department of Pharmacology, Penn State College of Medicine, 500 University Drive, Hershey, PA 17033

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Proteome Science 2010, 8:61  doi:10.1186/1477-5956-8-61

Published: 22 November 2010

Abstract

Background

Protein biomarkers will play a pivotal role in the future of personalized medicine for both diagnosis and treatment decision-making. While the results of several pre-clinical and small-scale clinical studies have demonstrated the value of protein biomarkers, there have been significant challenges to translating these findings into routine clinical care. Challenges to the use of protein biomarkers include inter-sample variability introduced by differences in post-collection handling and ex vivo degradation of proteins and protein modifications.

Results

In this report, we re-create laboratory and clinical scenarios for sample collection and test the utility of a new tissue stabilization technique in preserving proteins and protein modifications. In the laboratory setting, tissue stabilization with the Denator Stabilizor T1 resulted in a significantly higher yield of phospho-protein when compared to standard snap freeze preservation. Furthermore, in a clinical scenario, tissue stabilization at collection resulted in a higher yield of total phospho-protein, total phospho-tyrosine, pErkT202/Y204 and pAktS473 when compared to standard methods. Tissue stabilization did not have a significant effect on other post-translational modifications such as acetylation and glycosylation, which are more stable ex-vivo. Tissue stabilization did decrease total RNA quantity and quality.

Conclusion

Stabilization at the time of collection offers the potential to better preserve tissue protein and protein modification levels, as well as reduce the variability related to tissue processing delays that are often associated with clinical samples.