Summary
This chapter covers the various methods of mechanical cell disruption and tissue homogenization that are currently commercially available for processing minute samples (<1 ml) to larger production quantities. These mechanical methods of lysing do not introduce chemicals or enzymes to the system. However, the energies needed when using these “harsh” methods can be high and destroy the very proteins being sought.
The destruction of cell membranes and walls is effected by subjecting the cells (1) to shearing by liquid flow, (2) to exploding by pressure differences between inside and outside of cell, (3) to collision forces by impact of beads or paddles, or (4) a combination of these forces. Practical suggestions to optimize each method, where to acquire such equipment, and links to reference sources are included.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Reinkemeier, M., Rocken, W., and Leitzmann, C. (1996) A rapid mechanical lysing procedure for routine analysis of plasmids from lactobacilli, isolated from sourdoughs. Int J Food Microbiol 29, 93–104.
Rezwan, M., Lanéelle, M. A., Sander, P., and Daffé, M. (2007) Breaking down the wall: Fractionation of mycobacteria. J Microbiol Methods, 68(1), 32–9.
Jung, J., Xing, X., and Matsumoto, K. (2001) Kinetic analysis of disruption of excess activated sludge by Dyno Mill and characteristics of protein release for recovery of useful materials. Biochem. Eng. J. 8, 1–7.
Lizotte, E., Tremblay, A., Allen, B. G., and Fiset, C. (2005) Isolation and characterization of subcellular protein fractions from mouse heart. Anal Biochem 345, 47–54.
Lindeskog, P., Haaparanta, T., Norgard, M., Glaumann, H., Hansson, T., and Gustafsson, J. A. (1986) Isolation of rat intestinal microsomes: partial characterization of mucosal cytochrome P-450. Arch Biochem Biophys 244, 492–501.
Nally, J. E., Whitelegge, J. P., Aguilera, R., Pereira, M. M., Blanco, D. R., and Lovett, M. A. (2005) Purification and proteomic analysis of outer membrane vesicles from a clinical isolate of Leptospira interrogans serovar Copenhageni. Proteomics 5, 144–152.
Autuori, F., Brunk, U., Peterson, E., and Dallner, G. (1982) Fractionation of isolated liver cells after disruption with a nitrogen bomb and sonication. J Cell Sci 57, 1–13.
Kelly, W. J., and Muske, K. R. (2004) Optimal operation of high-pressure homogenization for intracellular product recovery. Bioprocess Biosyst Eng 27, 25–37.
Mehlhorn, I., Groth, D., Stockel, J., Moffat, B., Reilly, D., Yansura, D., Willett, W. S., Baldwin, M., Fletterick, R., Cohen, F. E., Vandlen, R., Henner, D., and Prusiner, S. B. (1996) High-level expression and characterization of a purified 142-residue polypeptide of the prion protein. Biochemistry 35, 5528–5537.
Vinatier, J., Herzog, E., Plamont, M. A., Wojcik, S. M., Schmidt, A., Brose, N., Daviet, L., El Mestikawy, S., and Giros, B. (2006) Interaction between the vesicular glutamate transporter type 1 and endophilin A1, a protein essential for endocytosis. J Neurochem 97, 1111–25.
Jaki, B. U., Franzblau, S. G., Cho, S. H., and Pauli, G. F. (2006) Development of an extraction method for mycobacterial metabolome analysis. J Pharm Biomed Anal 41, 196–200.
Acknowledgments
Superb assistance was rendered by the following people who are most conversant in their noted equipment areas. Bead milling by Tim Hopkins (BIOSPEC PRODUCTS) and Norbert Roskosch (W. A. BACHOFEN AG); Rotor/Stators by Roger Munsinger (KINEMATICA USA/AG); High-pressure flowing liquid by William Pandolfe (INVESYS) and Thomai Panagiotou (MICROFLUIDICS); High-pressure batch gas by Deanna Shepard (PARR INSTRUMENTS); Low-pressure flowing gas by Lee Clark (GLAS-COL); and ultrasonic processors by Marc Lusting and Andrea Coppola (MISONIX).
My book editor Dr. Anthon Posch has been exceptionally supportive in both technical as well as motivational matters. My wife Robin and two sons Evan and Adam are the sparkle of my life.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Goldberg, S. (2008). Mechanical/Physical Methods of Cell Disruption and Tissue Homogenization. In: Posch, A. (eds) 2D PAGE: Sample Preparation and Fractionation. Methods in Molecular Biology™, vol 424. Humana Press. https://doi.org/10.1007/978-1-60327-064-9_1
Download citation
DOI: https://doi.org/10.1007/978-1-60327-064-9_1
Publisher Name: Humana Press
Print ISBN: 978-1-58829-722-8
Online ISBN: 978-1-60327-064-9
eBook Packages: Springer Protocols