Project: Novel device for standardized and rapid blood Plasma Separation and STabilization (PSST)
Life science research, development and diagnostics rely heavily on blood-based analysis of proteins, peptides and metabolites. Most assays are performed on blood plasma and separation of plasma from whole blood is routinely done through centrifugation at animal house facilities and central laboratories. _x000D__x000D_Protein biomarkers are considered to have the potential to revolutionize diagnostics for cancer-, cardiovascular- and neurodegenerative diseases. However, a large obstacle for efficient strategies to find protein, peptide or metabolite biomarkers in body fluids is the lack of a standardized method for rapid separation and stabilization. The most commonly used method for separation, centrifugation, takes at least 10 minutes and requires relatively large volumes of blood samples which is in direct contrast to the need of current state-of-the-art analysis equipment. During this time, degradation of the blood samples has been ongoing for several minutes resulting in a substantial change in sample composition. This makes the subsequent analysis much more difficult or even impossible. The problem is widely published, for example in Ayache A et al., (Clinical Chemistry, 2006) and Boyanton B, “, (General Clinical Chemistry, 2002)._x000D__x000D_Together with Uppsala University, Denator AB has shown that several proteins and peptides that earlier have been reported to be related to specific disease states and have been suggested as pharmaceutical targets or biomarkers, are degraded within minutes after sampling (Sköld K et al. The significance of biochemical and molecular sample integrity in brain proteomics and peptidomics: Peptides as sample quality indicators”, in Proteomics 2007, 7, (24), 4445-56). Based on this research, Denator AB has developed a novel method to stabilize proteins, peptides and their modifications through rapid and homogenous heat inactivation. The methodology has been developed and launched for tissue samples and a proof of principle for the same method of stabilization of blood plasma has been generated. _x000D__x000D_The consortia intend to tackle the problem of poor quality blood plasma by developing and verifying a novel device for standardized and rapid blood plasma separation and stabilization, thereby offering a unique solution to an existing large problem in the life science research area as well as within clinical diagnostics. _x000D__x000D_The constellation of the consortia ensures that all key competencies required to develop and verify such a device are collected; _x000D_- Denator will utilize its unique knowledge in sample preparation, patents and instrumentation to develop and verify the stabilization part of the device. _x000D_- Sigolis has a unique combination of development skills and production capabilities within microstructures, handling of biological fluids in microsystems and experience from separation applications which will be utilized to develop the separation part of the device. _x000D_- Leiden University Medical Center (LUMC) will utilize their state-of-the-art laboratory and experience from plasma analysis to support and verify the development of the new device and in a second stage utilize it for biomarker research and publish the results. _x000D_- BioScope supports the development with state of the art knowledge of sample preparation protocols for efficient protein analysis. _x000D_- AstraZeneca ensures that the demands from large pharmaceutical companies are recognized in the design phase. _x000D__x000D_The goal is to develop and verify a fully functional prototype for blood samples in which plasma is rapidly separated from whole blood using microstructures, followed by stabilization using Denator's proprietary method for complete enzyme inactivation (i. e. stabilization). _x000D__x000D_The prototype will be further developed to a product which can be produced and sold within 12 months after the project is finalized. Initial target segment is protein science applications within academia and the pharmaceutical industry. The device ensures high quality blood plasma without disturbance from enzymatic degradation and enables efficient biomarker discovery and drug discovery research. _x000D__x000D_The device has the potential to be the future standard method for blood plasma handling for diagnostics and thereby enable a new generation of protein biomarkers to be implemented without today’s problem of sample degradation. The life science industry would thereby take a large step toward the vision of personalized medicine. _x000D_
| Acronym | PSST (Reference Number: 4816) |
| Duration | 01/04/2009 - 01/11/2010 |
| Project Topic | The purpose is to develop and verify a prototype for rapid and reliable blood plasma separation and stabilization thereby increasing the potential discovery of biomarkers for diagnostics and drug development. This will result in a commercially available disposable product. |
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Project Results (after finalisation) |
The PSST project consisted of seven work packages (WP's). The LUMC was responsible for WP1, namely "development, optimization and validation of plasma stabilization methodology", and responsible for WP5, termed "evaluation of plasma separation and stabilization prototype". To this end, plasma samples have been analyzed at the LUMC following a mass spectrometry (MS)-based proteomics strategy. The problem of degradation of proteins and peptides was demonstrated by comparing stabilized and untreated samples. Indeed, MS-analysis of stabilized plasma samples obtained with Stabilizor-technology showed better reproducibility in protein profiles. The stabilization parameters such as temeprature, heating time and buffer solutions have been optimized. |
| Network | Eurostars |
| Call | Eurostars Cut-Off 2 |
Project partner
| Number | Name | Role | Country |
|---|---|---|---|
| 5 | AstraZeneca R&D Mölndal | Partner | Sweden |
| 5 | Denator AB | Coordinator | Sweden |
| 5 | Departamento de Quimica, Faculdade de Ciências e Technologia, Universidade Nova de Lisboa | Partner | Portugal |
| 5 | Leiden University Medical Center, Department of Parasitology, Biomolecular Mass Spectrometry | Partner | Netherlands |
| 5 | Sigolis AB | Partner | Sweden |