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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wisnieskie JR, Zougman A, Nagaraj N, Mann M (2009) Universal sample preparation method for proteome analysis. Nat Methods 6(5):359–362
Wiśniewski JR, Ostasiewicz P, Mann M (2011) High recovery FASP applied to the proteomic analysis of microdissected formalin fixed paraffin embedded cancer tissues retrieve known colon cancer markers. J Proteome Res 10(7):3040–3049
Wiśniewski JR, Nagaraj N, Zougman A, Gnad F, Mann M (2010) Brain phosphoproteome obtained by a FASP-based method reveals plasma membrane protein topology. J Proteome Res 9(6):3280–3289
Erde J, Ogorzalek Loo RR, Loo JA (2014) Enhanced FASP (eFASP) to increase proteome coverage and sample recovery for quantitative proteomic experiments. J Proteome Res 13(4):1885–1895
Deeb SJ, Cox J, Schmidt-Supprian M, Mann M (2014) N-linked glycosylation enrichment for in-depth cell surface proteomics of diffuse large B-cell lymphoma subtypes. Mol Cell Proteomics 13(1):240–251
Hecht ES, McCord JP, Muddiman DC (2016) A quantitative glycomics and proteomics combined purification strategy. J Vis Exp 109:e53735. doi:10.3791/53735
Baycin-Hizal D, Tabb DL, Chaerkady R, Chen L, Lewis NE, Nagarajan H, Sarkaria V, Kumar A, Wolozny D, Colao J, Jacobsen E, Tian Y, O’Meally RN, Krag SS, Cole RN, Palsson BO, Zhang H, Betenbaugh M (2012) Proteomic analysis of Chinese hamster ovary cells. J Proteome Res 11(11):5265–5272
Acknowledgments
The authors acknowledge funding from Science Foundation Ireland (grant ref. 13/1A/1841).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Coleman, O., Henry, M., Clynes, M., Meleady, P. (2017). Filter-Aided Sample Preparation (FASP) for Improved Proteome Analysis of Recombinant Chinese Hamster Ovary Cells. In: Meleady, P. (eds) Heterologous Protein Production in CHO Cells. Methods in Molecular Biology, vol 1603. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6972-2_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6972-2_12
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6971-5
Online ISBN: 978-1-4939-6972-2
eBook Packages: Springer Protocols