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Biotechnology and Bioprocess Engineering

, Volume 13, Issue 5, pp 613–623 | Cite as

The performance of a glass bead shaking technique for the disruption of Escherichia coli cells

  • Ramakrishnan Nagasundara Ramanan
  • Tau Chuan Ling
  • Arbakariya B. Ariff
Article
First Online:
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Accepted:

Abstract

The efficacy of a simple laboratory method for cell disruption based on the shaking of glass beads on a rotary shaker was assessed in this study, via measurements of the release of total protein and interferon-α2b from E. coli. The optimum conditions for cell disruption were detected after 30 min of shaking in Tris-HCl buffer (pH 8) at 300 rpm with 1.5 g of glass beads (diameter: 0.5 mm) per mL of cell suspension volume. Three test runs were conducted under the above conditions and the maximum average protein release values were determined as 3.048, 3.564, and 3.015 mg/mL, respectively. The amount of protein release was comparable to the amount of protein release in ultrasonica-tion and glass bead vortexing procedures. The amount of interferon-α2b release in the ultrasonication, glass bead vortexing, and glass bead shaking trials were 240, 172, and 201 ng/mL, respectively. This method was shown to process between 1 and 10 mL of sample volume in a 50 mL Falcon tube without a great deal of deviation, and was able to handle in excess of 60 samples simultaneously.

Keywords

cell disruption downstream processing E. coli glass beads shaker interferon-α2b 
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Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg GmbH 2008

Authors and Affiliations

  • Ramakrishnan Nagasundara Ramanan
    • 1
  • Tau Chuan Ling
    • 2
  • Arbakariya B. Ariff
    • 1
    • 3
  1. 1.Institute of BioscienceUniversiti Putra MalaysiaSerdangMalaysia
  2. 2.Department of Process and Food Engineering, Faculty of EngineeringUniversiti Putra MalaysiaSerdangMalaysia
  3. 3.Department of Bioprocess Technology, Faculty of Biotechnology and Biomolecular SciencesUniversiti Putra MalaysiaSerdangMalaysia

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