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      Stability of Proteins in Dried Blood Spot Biobanks*

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          Abstract

          An important motivation for the construction of biobanks is to discover biomarkers that identify diseases at early, potentially curable stages. This will require biobanks from large numbers of individuals, preferably sampled repeatedly, where the samples are collected and stored under conditions that preserve potential biomarkers. Dried blood samples are attractive for biobanking because of the ease and low cost of collection and storage. Here we have investigated their suitability for protein measurements. Ninety-two proteins with relevance for oncology were analyzed using multiplex proximity extension assays (PEA) in dried blood spots collected on paper and stored for up to 30 years at either +4 °C or −24 °C.

          Our main findings were that (1) the act of drying only slightly influenced detection of blood proteins (average correlation of 0.970), and in a reproducible manner (correlation of 0.999), (2) detection of some proteins was not significantly affected by storage over the full range of three decades (34 and 76% of the analyzed proteins at +4 °C and −24 °C, respectively), whereas levels of others decreased slowly during storage with half-lives in the range of 10 to 50 years, and (3) detectability of proteins was less affected in dried samples stored at −24 °C compared with at +4 °C, as the median protein abundance had decreased to 80 and 93% of starting levels after 10 years of storage at +4 °C or −24 °C, respectively. The results of our study are encouraging as they suggest an inexpensive means to collect large numbers of blood samples, even by the donors themselves, and to transport, and store biobanked samples as spots of whole blood dried on paper. Combined with emerging means to measure hundreds or thousands of protein, such biobanks could prove of great medical value by greatly enhancing discovery as well as routine analysis of blood biomarkers.

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          Most cited references31

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          What a drop can do: dried blood spots as a minimally invasive method for integrating biomarkers into population-based research.

          Logistical constraints associated with the collection and analysis of biological samples in community-based settings have been a significant impediment to integrative, multilevel bio-demographic and biobehavioral research. However recent methodological developments have overcome many of these constraints and have also expanded the options for incorporating biomarkers into population-based health research in international as well as domestic contexts. In particular using dried blood spot (DBS) samples-drops of whole blood collected on filter paper from a simple finger prick-provides a minimally invasive method for collecting blood samples in nonclinical settings. After a brief discussion of biomarkers more generally, we review procedures for collecting, handling, and analyzing DBS samples. Advantages of using DBS samples-compared with venipuncture include the relative ease and low cost of sample collection, transport, and storage. Disadvantages include requirements for assay development and validation as well as the relatively small volumes of sample. We present the results of a comprehensive literature review of published protocols for analysis of DBS samples, and we provide more detailed analysis of protocols for 45 analytes likely to be of particular relevance to population-level health research. Our objective is to provide investigators with the information they need to make informed decisions regarding the appropriateness of blood spot methods for their research interests.
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            Therapeutic Drug Monitoring by Dried Blood Spot: Progress to Date and Future Directions

            This article discusses dried blood spot (DBS) sampling in therapeutic drug monitoring (TDM). The most important advantages of DBS sampling in TDM are the minimally invasive procedure of a finger prick (home sampling), the small volume (children), and the stability of the analyte. Many assays in DBS have been reported in the literature over the previous 5 years. These assays and their analytical techniques are reviewed here. Factors that may influence the accuracy and reproducibility of DBS methods are also discussed. Important issues are the correlation with plasma/serum concentrations and the influence of hematocrit on spot size and recovery. The different substrate materials are considered. DBS sampling can be a valid alternative to conventional venous sampling. However, patient correlation studies are indispensable to prove this. Promising developments are dried plasma spots using membrane and hematocrit correction using the potassium concentration.
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              Micro-scale blood plasma separation: from acoustophoresis to egg-beaters.

              Plasma is a rich mine of various biomarkers including proteins, metabolites and circulating nucleic acids. The diagnostic and therapeutic potential of these analytes has been quite recently uncovered, and the number of plasma biomarkers will still be growing in the coming years. A significant part of the blood plasma preparation is still handled manually, off-chip, via centrifugation or filtration. These batch methods have variable waiting times, and are often performed under non-reproducible conditions that may impair the collection of analytes of interest, with variable degradation. The development of miniaturised modules capable of automated and reproducible blood plasma separation would aid in the translation of lab-on-a-chip devices to the clinical market. Here we propose a systematic review of major plasma analytes and target applications, alongside existing solutions for micro-scale blood plasma extraction, focusing on the approaches that have been biologically validated for specific applications.
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                Author and article information

                Journal
                Mol Cell Proteomics
                Mol. Cell Proteomics
                mcprot
                mcprot
                MCP
                Molecular & Cellular Proteomics : MCP
                The American Society for Biochemistry and Molecular Biology
                1535-9476
                1535-9484
                July 2017
                13 May 2017
                13 May 2017
                : 16
                : 7
                : 1286-1296
                Affiliations
                [1]From the ‡Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden;
                [2]§Danish Center for Neonatal Screening, Statens Serum Institut, Copenhagen, Denmark;
                [3]¶Centre for Inherited Metabolic Diseases, Karolinska University Hospital, Stockholm, Sweden;
                [4]‖Department of Public Health and Caring Sciences, Uppsala University, Uppsala, Sweden;
                [5]**Department of Medical Sciences, Uppsala University, Uppsala, Sweden
                Author notes
                ‡‡ To whom correspondence should be addressed: To whom the correspondence should be addressed: ulf.landegren@ 123456igp.uu.se .
                Article
                RA117.000015
                10.1074/mcp.RA117.000015
                5500761
                28501802
                91a49e8b-c45f-43b8-837a-c006bc6aeb4e
                © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

                Author's Choice—Final version free via Creative Commons CC-BY license.

                History
                : 7 April 2017
                : 18 May 2017
                Funding
                Funded by: Swedish Research Council
                Award ID: VR-INFRA 2009–6285
                Funded by: Swedish Research Council
                Award ID: VR-NT 2013–06023
                Funded by: European Research Council under the European Union's Seventh Framework Programme
                Award ID: (FP/2007–2013)/ERC Grant Agreement No 294409 (ProteinSeq)
                Funded by: IngaBritt och Arne Lundbergs Forskningsstiftelse (Ingabritt and Arne Lundberg Research Foundation)
                Funded by: Marie Curie ITN
                Award ID: (FP7/2007–2013) Grant Agreement No 316929 (GastricGlycoExplorer)
                Funded by: Danish National Biobank resource
                Award ID: supported by the Novo Nordisk Foundation
                Categories
                Technological Innovation and Resources

                Molecular biology
                Molecular biology

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