A Cost-Effective High-Throughput Plasma and Serum Proteomics Workflow Enables Mapping of the Molecular Impact of Total Pancreatectomy with Islet Autotransplantation
| Autor: | Gregory J. Beilman, Ofer Levy, Melena D. Bellin, Yue Xuan, Darwin L. Conwell, Zobeida Cruz-Monserrate, Hanno Steen, Judith A. Steen, Vibeke Andersen, Frederik Trier Møller, Allan Stensballe, Tue Bjerg Bennike |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Proteomics
0301 basic medicine Cost-Benefit Analysis medicine.medical_treatment Islets of Langerhans Transplantation Computational biology Blood Proteins/analysis Tandem mass spectrometry Transplantation Autologous Biochemistry Article Workflow 03 medical and health sciences Pancreatectomy proteomics Tandem Mass Spectrometry medicine data-independent acquisition Humans Data-independent acquisition plasma business.industry TPIAT Blood Proteins General Chemistry Blood proteins Autotransplantation 030104 developmental biology Pancreatitis Biomarker (medicine) biomarker Proteomics/methods business serum Biomarkers Biomarkers/blood Chromatography Liquid |
| Zdroj: | Bennike, T B, Bellin, M D, Xuan, Y, Stensballe, A, Møller, F T, Beilman, G J, Levy, O, Cruz-Monserrate, Z, Andersen, V, Steen, J, Conwell, D L & Steen, H 2018, ' A Cost-Effective High-Throughput Plasma and Serum Proteomics Workflow Enables Mapping of the Molecular Impact of Total Pancreatectomy with Islet Autotransplantation ', Journal of Proteome Research, vol. 17, no. 5, pp. 1983-1992 . https://doi.org/10.1021/acs.jproteome.8b00111 Bennike, T B, Bellin, M D, Xuan, Y, Stensballe, A, Trier Møller, F, Beilman, G J, Levy, O, Cruz-Monserrate, Z, Andersen, V, Steen, J, Conwell, D L & Steen, H 2018, ' A cost-effective high throughput plasma and serum proteomics workflow enables mapping of the molecular impact of total pancreatectomy with islet autotransplantation ', Journal of Proteome Research, vol. 17, no. 5, pp. 1983–1992 . https://doi.org/10.1021/acs.jproteome.8b00111 |
| DOI: | 10.1021/acs.jproteome.8b00111 |
| Popis: | Blood is an ideal body fluid for the discovery or monitoring of diagnostic and prognostic protein biomarkers. However, discovering robust biomarkers requires the analysis of large numbers of samples to appropriately represent interindividual variability. To address this analytical challenge, we established a high-throughput and cost-effective proteomics workflow for accurate and comprehensive proteomics at an analytical depth applicable for clinical studies. For validation, we processed one μL each from 62 plasma samples in 96-well plates and analyzed the product by quantitative data-independent acquisition (DIA) liquid chromatography/mass spectrometry (LC/MS); the data were queried using feature quantification with Spectronaut. To show the applicability of our workflow to serum, we analyzed a unique set of samples from 48 chronic pancreatitis patients, pre- and post Total Pancreatectomy with Islet AutoTransplantation (TPIAT) surgery. We identified 16 serum proteins with a statistically significant abundance alteration, representing a molecular signature distinct from that of chronic pancreatitis. In summary, we established a cost-efficient high throughput workflow for comprehensive proteomics using PVDF-membrane-based digestion that is robust, automatable, and applicable to small plasma- and serum volumes, e.g. fingerstick. Application of this plasma/serum proteomics workflow resulted in the first mapping of the molecular implications of TPIAT on the serum proteome. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|