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Filter-Aided Sample Preparation (FASP) for Improved Proteome Analysis of Recombinant Chinese Hamster Ovary Cells

  • Orla ColemanEmail author
  • Michael Henry
  • Martin Clynes
  • Paula Meleady
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1603)

Abstract

Chinese hamster ovary (CHO) cells are the most commonly used mammalian host cell line for biopharmaceutical production because of their ability to correctly fold and posttranslationally modify recombinant proteins that are compatible with human use. Proteomics, along with other ‘omic platforms, are being used to understand the biology of CHO cells with the ultimate aim of enhancing CHO cell factories for more efficient production of biopharmaceuticals. In this chapter, we will describe an efficient protocol called Filter Aided Sample Preparation (FASP) for the extraction of proteins from CHO cells for proteomic studies. FASP uses a common ultrafiltration device whereby the membrane pores are small enough to allow contaminating detergents to pass through, while proteins are too large and are retained and concentrated in the filter unit. This method of sample preparation and protein digestion is universally applicable and can be easily employed in any proteomics facilities as standard everyday laboratory reagents and equipment are used.

Key words

Proteomics Protein solubilization Filter-aided sample preparation FASP Chinese hamster ovary LC-MS/MS 

Notes

Acknowledgments

The authors acknowledge funding from Science Foundation Ireland (grant ref. 13/1A/1841).

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Orla Coleman
    • 1
    Email author
  • Michael Henry
    • 1
  • Martin Clynes
    • 1
  • Paula Meleady
    • 1
  1. 1.National Institute for Cellular BiotechnologyDublin City UniversityDublin 9Ireland

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