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Related Experiment Video

Updated: Jul 6, 2026

Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications
09:22

Budding Yeast Protein Extraction and Purification for the Study of Function, Interactions, and Post-translational Modifications

Published on: October 30, 2013

A microscale yeast cell disruption technique for integrated process development strategies.

Marc D Wenger1, Peter DePhillips, Daniel G Bracewell

  • 1The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK. marc_wenger@merck.com

Biotechnology Progress
|April 16, 2008
PubMed
Summary
This summary is machine-generated.

Adaptive Focused Acoustics (AFA) effectively disrupts yeast cells for microscale protein purification. This noncontact method yields results comparable to homogenization, accelerating process development for intracellular proteins.

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Area of Science:

  • Biotechnology
  • Bioprocess Engineering
  • Protein Purification

Background:

  • Miniaturizing protein purification accelerates development through parallel experimentation and automation.
  • Intracellular protein expression in yeast requires efficient small-scale cell disruption methods compatible with large-scale techniques.
  • Current methods need to provide representative protein release and contaminant profiles for downstream process studies.

Purpose of the Study:

  • Optimize adaptive focused acoustics (AFA) for disrupting milligram quantities of yeast cells.
  • Evaluate AFA for the microscale purification of recombinant human papillomavirus (HPV) virus-like particles (VLPs).
  • Compare AFA-based disruption to laboratory-scale homogenization for protein release and VLP recovery.

Main Methods:

  • Adaptive Focused Acoustics (AFA) was optimized for yeast cell disruption.
  • Microscale chromatography was used for subsequent VLP purification.
  • Yeast lytic enzyme was tested in combination with AFA for improved disruption efficiency.
  • Cell disruption and VLP recovery were assessed using light microscopy and chromatographic analysis.

Main Results:

  • AFA achieved soluble protein release equivalent to laboratory-scale homogenization.
  • VLP recovery using AFA followed by microscale chromatography was within 10% of homogenization, with comparable purity.
  • Addition of yeast lytic enzyme reduced processing time by nearly 3-fold and enhanced lysate comparability.
  • AFA minimized sample heating (< =8°C increase), unlike conventional sonication.

Conclusions:

  • AFA is a viable noncontact method for efficient yeast cell disruption at the microscale.
  • AFA combined with microscale chromatography enables rapid process development for intracellularly expressed proteins.
  • This approach facilitates representative studies for downstream purification of recombinant proteins.