A Cost-Effective
High-Throughput Plasma and Serum
Proteomics Workflow Enables Mapping of the Molecular Impact of Total
Pancreatectomy with Islet Autotransplantation
Version 2 2018-04-20, 19:47
Version 1 2018-04-19, 13:50
Posted on 2018-04-20 - 19:47
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 1 μL each from 62 plasma samples in 96-well plates
and analyzed the product by quantitative data-independent acquisition
liquid chromatography/mass spectrometry; 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 statistically significant abundance
alterations, which represent 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., finger stick. Application of this plasma/serum
proteomics workflow resulted in the first mapping of the molecular
implications of TPIAT on the serum proteome.
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Bennike, Tue Bjerg; D. Bellin, Melena; Xuan, Yue; Stensballe, Allan; Møller, Frederik Trier; J. Beilman, Gregory; et al. (2018). A Cost-Effective
High-Throughput Plasma and Serum
Proteomics Workflow Enables Mapping of the Molecular Impact of Total
Pancreatectomy with Islet Autotransplantation. ACS Publications. Collection. https://doi.org/10.1021/acs.jproteome.8b00111
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AUTHORS (12)
TB
Tue Bjerg Bennike
MD
Melena D. Bellin
YX
Yue Xuan
AS
Allan Stensballe
FM
Frederik Trier Møller
GJ
Gregory J. Beilman
OL
Ofer Levy
ZC
Zobeida Cruz-Monserrate
VA
Vibeke Andersen
JS
Judith Steen
DC
Darwin L. Conwell
HS
Hanno Steen
KEYWORDS
body fluidfeature quantification16 serum proteins1 μ Lpancreatitis patientshigh-throughput workflowMolecular Impactproteomics workflowTPIATserum proteome62 plasma samplesprognostic protein biomarkersinterindividual variabilityIslet Autotransplantation Bloodserum volumesSerum Proteomics Workflow Enables MappingPVDF-membrane-based digestiondata-independent acquisitionTotal PancreatectomyCost-Effective High-Throughput Plasmaabundance alterationsislet autotransplantation