Standardized and automated solid-phase extraction procedures for high-throughput proteomics of body fluids

J Proteomics. 2012 Dec 21:77:144-53. doi: 10.1016/j.jprot.2012.07.023. Epub 2012 Jul 26.

Abstract

In order to balance the speed of analytical sample preparation procedures with mass spectrometry (MS)-based clinical proteomics the application of high-throughput robotic systems for body fluid workup is essential. In this paper we describe the implementation of various solid-phase extraction (SPE) sample preparation protocols on two different platforms, namely: 1) Magnetic bead-based SPE of peptides and proteins from body fluids on a Hamilton liquid handling workstation; 2) Cartridge-based SPE on a SPARK Symbiosis system. All SPE protocols were optimized for MS-based proteomics and compared with respect to obtained peptide- and protein profiles. Throughput numbers that were achieved in a 24 hour time frame for the sample workup procedures were more than 700 samples for the magnetic bead-based method and over 1000 samples for the cartridge-based method.

MeSH terms

  • Blood Proteins / chemistry*
  • Blood Proteins / isolation & purification*
  • Blood Proteins / metabolism
  • Female
  • Humans
  • Male
  • Plasma / chemistry*
  • Plasma / metabolism
  • Proteomics / instrumentation*
  • Proteomics / methods*
  • Proteomics / standards*
  • Robotics / instrumentation
  • Robotics / methods

Substances

  • Blood Proteins