Automated Sample Preparation Method for Mass Spectrometry Analysis on Recombinant Proteins

J Chromatogr A. 2009 May 15;1216(20):4457-64. doi: 10.1016/j.chroma.2009.03.041. Epub 2009 Mar 20.

Abstract

A completely automated procedure for the purification and desalting of proteins with a polyhistidine purification tag prior to mass spectrometry analysis is presented. The system is ideal for rapid quality control and optimization studies and it provides researchers with a straightforward, reliable tool for studies of recombinant proteins. Forty-eight samples can be prepared within 4.5h and only small cultivation and buffer volumes are needed. In this proof of concept, 19,000-35,000Da recombinant proteins from both crude and clarified cell lysates were successfully prepared for subsequent analysis by electrospray ionization and matrix-assisted laser desorption/ionization mass spectrometry as well as by gel electrophoresis.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Automation
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Humans
  • Recombinant Proteins / chemistry*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Recombinant Proteins