+ Site Statistics
References:
54,258,434
Abstracts:
29,560,870
PMIDs:
28,072,757
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Comprehensive and Scalable Highly Automated MS-Based Proteomic Workflow for Clinical Biomarker Discovery in Human Plasma



Comprehensive and Scalable Highly Automated MS-Based Proteomic Workflow for Clinical Biomarker Discovery in Human Plasma



Journal of Proteome Research 2014



Over the past decade, mass spectrometric performance has greatly improved in terms of sensitivity, dynamic range, and speed. By contrast, only limited progress has been accomplished with regard to automation, throughput, and robustness of the proteomic sample preparation process upstream of mass spectrometry. The present work delivers an optimized analysis of human plasma samples in both small preclinical and large clinical studies, enabled by the development of a highly automated quantitative proteomic workflow. Several iterative evaluation and validation steps were performed before process "design freeze" and development completion. A robotic liquid handling workflow and platform (including reduction, alkylation, digestion, TMT labeling, pooling, and purification) were shown to provide better quantitative trueness and precision than manual operation at the bench. Depletion of the most abundant human plasma proteins and subsequent buffer exchange were also developed and integrated. Finally, 96 identical pooled human plasma samples were prepared in a 96-well plate format, and each sample was individually subjected to our developed workflow. This test revealed increased throughput and robustness compared with to-date published manual or less automated workflows. Our workflow is ready-to-use for future (pre-) clinical studies. We expect our work to facilitate, accelerate, and improve clinical proteomic discovery in human blood plasma.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 052269946

Download citation: RISBibTeXText

PMID: 25058407

DOI: 10.1021/pr500635f


Related references

A Highly Automated Shotgun Proteomic Workflow: Clinical Scale and Robustness for Biomarker Discovery in Blood. Methods in Molecular Biology 1619: 433-449, 2018

A comprehensive workflow of mass spectrometry-based untargeted metabolomics in cancer metabolic biomarker discovery using human plasma and urine. Metabolites 3(3): 787-819, 2013

Differential gel-based proteomic approach for cancer biomarker discovery using human plasma. Methods in Molecular Biology 854: 223-237, 2012

Proteomic data analysis workflow for discovery of candidate biomarker peaks predictive of clinical outcome for patients with acute myeloid leukemia. Journal of Proteome Research 7(6): 2332-2341, 2008

Fast quantitative urinary proteomic profiling workflow for biomarker discovery in kidney cancer. Clinical Proteomics 15: 42-42, 2019

Utilizing human blood plasma for proteomic biomarker discovery. Journal of Proteome Research 4(4): 1073-1085, 2005

Proteomic profiling of human plasma for cancer biomarker discovery. Proteomics 17(6), 2016

A proteomic workflow for discovery of serum carrier protein-bound biomarker candidates of alcohol abuse using LC-MS/MS. Electrophoresis 30(12): 2207-2214, 2009

Toward Biomarker Development in Large Clinical Cohorts: An Integrated High-Throughput 96-Well-Plate-Based Sample Preparation Workflow for Versatile Downstream Proteomic Analyses. Analytical Chemistry 88(17): 8518-8525, 2018

Proteomic Biomarker Discovery in 1000 Human Plasma Samples with Mass Spectrometry. Journal of Proteome Research 15(2): 389-399, 2016

Computational biomarker pipeline from discovery to clinical implementation: plasma proteomic biomarkers for cardiac transplantation. Plos Computational Biology 9(4): E1002963, 2013

Proteomics of Cerebrospinal Fluid: Throughput and Robustness Using a Scalable Automated Analysis Pipeline for Biomarker Discovery. Analytical Chemistry 87(21): 10755-10761, 2016

Time course proteomic profiling of human myocardial infarction plasma samples: an approach to new biomarker discovery. Clinica Chimica Acta; International Journal of Clinical Chemistry 412(11-12): 1086-1093, 2012

Isolation of urinary exosomes for RNA biomarker discovery using a simple, fast, and highly scalable method. Methods in Molecular Biology 1182: 145-170, 2015

Solvent interaction analysis as a proteomic approach to structure-based biomarker discovery and clinical diagnostics. Expert Review of Proteomics 13(1): 9-17, 2016