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Use of focused acoustics for cell disruption to provide ultra scale-down insights of microbial homogenisation and its bioprocess impact - recovery of antibody fragments from rec E. coli.
MedLine Citation:
PMID:  22383367     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
An ultra scale-down device that provides insight of how industrial homogenisation impacts bioprocess performance is desirable in the biopharmaceutical industry, especially at the early stage of process development where only a small quantity of material is available. In this work, we assess the effectiveness of focused acoustics as the basis of an ultra scale-down cell disruption method to mimic and study high-pressure, step-wise homogenisation of rec E. coli cells for the recovery of an intracellular protein, antibody fragment (Fab'). The release of both Fab' and of overall protein follows first order reaction kinetics with respect to time of exposure to focused acoustics. The rate constant is directly proportional to applied electrical power input per unit volume. For nearly total protein or Fab' release (>99%), the key physical properties of the disruptate produced by focused acoustics, such as cell debris particle size distribution and apparent viscosity show good agreement with those for homogenates produced by high pressure homogenisation operated to give the same fractional release. The only key difference is observed for partial disruption of cells where focused acoustics yields a disruptate of lower viscosity than homogenisation, evidently due to a greater extent of polynucleic acids degradation. Verification of this ultra scale-down approach to cell disruption by high pressure homogenisation is achieved using ultra scale-down centrifugation to demonstrate the same sedimentation characteristics of disruptates prepared using both the scaled-down focused acoustic and the pilot-scale homogenisation methods for the same fraction of protein release. Biotechnol. Bioeng. © 2012 Wiley Periodicals, Inc.
Authors:
Qiang Li; Jean P Aucamp; Alison Tang; Alex Chatel; Mike Hoare
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-3-1
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  -     ISSN:  1097-0290     ISO Abbreviation:  -     Publication Date:  2012 Mar 
Date Detail:
Created Date:  2012-3-2     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
Copyright © 2012 Wiley Periodicals, Inc.
Affiliation:
Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK. qli@hotmail.co.uk.
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