title: 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.
abstract: An ultra scale-down (USD) device that provides insight of how industrial homogenization 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 USD cell disruption method to mimic and study high-pressure, step-wise homogenization of rec Escherichia 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 homogenization 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 homogenization, evidently due to a greater extent of polynucleic acids degradation. Verification of this USD approach to cell disruption by high-pressure homogenization is achieved using USD centrifugation to demonstrate the same sedimentation characteristics of disruptates prepared using both the scaled-down focused acoustic and the pilot-scale homogenization methods for the same fraction of protein release.
authors.affiliation: Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, UK. qli@hotmail.co.uk
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.forename: Jean P
authors.initials: JP
authors.lastname: Aucamp
authors.forename: Alison
authors.initials: A
authors.lastname: Tang
authors.forename: Alex
authors.initials: A
authors.lastname: Chatel
authors.forename: Mike
authors.initials: M
authors.lastname: Hoare
mesh_terms: D001433:Bacteriolysis; D001709:Biotechnology / Q000379:methods*; D004560:Electricity; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics; D007128:Immunoglobulin Fragments / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000302:isolation & purification; D013016:Sound; D013678:Technology, Pharmaceutical / Q000379:methods
chemical_list: D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins