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Adaptive Focused Acoustics®(AFA®) for Cell Lysis

Cell lysis methods differ greatly depending on the species the cells are derived from. Covaris Adaptive Focused Acoustics®(AFA®) is unique in its ability to control the amount of energy delivered to samples making it ideal for sonication cell lysis workflows.

Contents

Robust extraction workflow solutions for cells lysis Powered by AFA®

Covaris AFA technology delivers highly tunable energy through Focused-ultrasonicator instruments. By controlling the dosage of acoustic energy delivered, it is possible to gently disrupt the cell membranes of mammalian cells (e.g., GPCR assays), or abruptly disrupt the cell walls of bacteria (e.g., total protein extraction). AFA provides the flexibility to homogenize cells and efficiently extract proteins, RNA, DNA, Metabolites, or other targets in buffers optimized for downstream analytical methods.

Downstream Multiomics Workflows:



Covaris Cell Lysis Workflows

A. Direct cell lysis

 

B. Dry tissue pulverization extraction workflow using cryoPREP and AFA

 

Resources

Cell Lysis Application Notes
Application notes to support your cell lysis workflows including mammalian cell lysis, bacterial cell lysis, HMW extraction and more.
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References

E.coli

  1. Li, Q., Mannall, G. J., Ali, S., & Hoare, M. (2013). An ultra scale-down approach to study the interaction of fermentation, homogenization, and centrifugation for antibody fragment recovery from recE. coli. Biotechnology and Bioengineering, 110(8), 2150-2160. doi:10.1002/bit.24891
  2. Li, Q., Aucamp, J. P., Tang, A., Chatel, A., & Hoare, M. (2012). Use of focused acoustics for cell disruption to provide ultra scale-down insights of microbial homogenization and its bioprocess impact-recovery of antibody fragments from rec E. coli. Biotechnology and Bioengineering, 109(8), 2059-2069. doi:10.1002/bit.24484
  3. Perez-Pardo, M. A., Ali, S., Balasundaram, B., Mannall, G. J., Baganz, F., & Bracewell, D. G. (2011). Assessment of the manufacturability of Escherichia coli high cell density fermentations. Biotechnology Progress, 27(5), 1488-1496. doi:10.1002/btpr.644
  4. Ordidge, G. C. Rapid microscale evaluation of the impact of fermentation conditions on inclusion body formation, solubilization and protein refolding yields (Unpublished doctoral dissertation). University College London (University of London).
  5. Release of Microbial Cells from a Solid Matrix, Cell Lysis, and Shearing of Nucleic Acids Performed in a One-Step Procedure

Mammalian Cell lysis

  1. Budnik, B., Levy, E., & Slavov, N. (2017). Mass-spectrometry of single mammalian cells quantifies proteome heterogeneity during cell differentiation. doi:10.7287/peerj.preprints.2767
  2. Dhabaria, A., Cifani, P., Reed, C., Steen, H., & Kentsis, A. (2015). A High-Efficiency Cellular Extraction System for Biological Proteomics. Journal of Proteome Research, 14(8), 3403-3408. doi:10.1021/acs.jproteome.5b00547
  3. Li, S., Plouffe, B. D., Belov, A. M., Ray, S., Wang, X., Murthy, S. K., . . . Ivanov, A. R. (2015). An Integrated Platform for Isolation, Processing, and Mass Spectrometry-based Proteomic Profiling of Rare Cells in Whole Blood. Molecular & Cellular Proteomics, 14(6), 1672-1683. doi:10.1074/mcp.m114.045724
  4. Ly, M., Lu, F., Maheshwari, G., & Subramanian, S. (2014). Microscale acoustic disruption of mammalian cells for intracellular product release. Journal of Biotechnology, 184, 146-153. doi:10.1016/j.jbiotec.2014.04.030

Yeast Cell Lysis

  1. Bláha BAF, Morris SA, Ogonah OW, et al. Development of a high-throughput microscale cell disruption platform for Pichia pastoris in rapid bioprocess design. Biotechnology Progress. 2017;34(1):130-140. doi:10.1002/btpr.2555.
  2. Wenger M, Dephillips P, Bracewell D. A Microscale Yeast Cell Disruption Technique for Integrated Process Development Strategies. Biotechnology Progress. 2008;24(3):606-614. doi:10.1021/bp070359s.
  3. Application Note: Active Extraction of Native Proteins from Yeast using Covaris Adaptive Focused Acoustics™ (AFA)
  4. Application Note: Protein Extraction from Yeast: Comparison of the Covaris Adaptive Focused Acoustics™ (AFA) Process to Conventional Bead Beating and Probe Sonication

Pseudomonas disruption

Application Note: Effective disruption of Pseudomonas aeruginosa biofilms using Adaptive Focused Acoustics™ (AFA)
Plant Cells
  1. Toorchi, M., Nouri, M., Tsumura, M., & Komatsu, S. (2008). Acoustic Technology for High-Performance Disruption and Extraction of Plant Proteins. Journal of Proteome Research, 7(7), 3035-3041. doi:10.1021/pr800012c
  2. Kota, U., & Goshe, M. B. (2011). Advances in qualitative and quantitative plant membrane proteomics. Phytochemistry, 72(10), 1040-1060. doi:10.1016/j.phytochem.2011.01.027
Hydrogels
  1. Burgess, K. A., Miller, A. F., Oceandy, D., & Saiani, A. (2017). Western blot analysis of cells encapsulated in self-assembling peptide hydrogels. BioTechniques, 63(6). doi:10.2144/000114617
CryoPREP
  1. Mundt, F., Rajput, S., Li, S., Ruggles, K. V., Mooradian, A. D., Mertins, P., . . . Ma, C. X. (2018). Mass spectrometry-based proteomics reveals potential roles of NEK9 and MAP2K4 in resistance to PI3K inhibitors in triple negative breast cancers. Cancer Research. doi:10.1158/0008-5472.can-17-1990
  2. Wylezich C, Papa A, Beer M, Höper D. A Versatile Sample Processing Workflow for Metagenomic Pathogen Detection. Sci Rep. 2018;8(1):13108.