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A robotic protocol for high‐throughput processing of samples for selected reaction monitoring assays
Author(s) -
Zhu Min,
Zhang Pingbo,
GengSpyropoulos Minghui,
Moaddel Ruin,
Semba Richard D.,
Ferrucci Luigi
Publication year - 2017
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201600339
Subject(s) - context (archaeology) , protocol (science) , sample preparation , reproducibility , throughput , chromatography , selected reaction monitoring , mass spectrometry , instrumentation (computer programming) , computer science , chemistry , tandem mass spectrometry , biology , medicine , operating system , pathology , paleontology , alternative medicine , wireless
Selected reaction monitoring mass spectrometry (SRM‐MS) is a sensitive and accurate method for the quantification of targeted proteins in biological specimens. However, the sample throughput and reliability of this technique is limited by the complexity of sample preparation, as well as instrumentation and data processing. Modern robotic equipment allows for rapid and accurate processing of large number of samples and makes SRM‐MS assay applicable in epidemiological studies. Herein, we describe an automated sample processing platform developed in the context of an SRM‐MS protocol for the assay of complement factor H protein and its variants in human plasma. We report detailed performance data on plasma digestion, sample cleanup and optimized robotic handling implemented on a Biomek ® NX p Workstation. Method validation was assessed with isotopically labeled peptide standards and had high reproducibility of intra‐day assay (CVs from 2.7 to 17.5% with median CV of 5.3%) and inter‐day assay (CVs from 4.8 to 17.6 with median CV of 7.2%) for all peptides.