Selective flocculation and precipitation for the improvement of virus-like particle recovery from yeast homogenate.
The purification of an intracellular product from a complex mixture of contaminants after cell disruption is a common problem in processes downstream of fermentation systems. This is particularly challenging for the recovery of particulate (80 nm in diameter) multimeric protein products, named virus-like particles (VLPs), from cell debris and other intracellular components. Selective flocculation for debris removal followed by selective precipitation of the target protein can be used as a preclarification step to aid purification. In this paper, selective borax flocculation of cell debris in yeast homogenate, followed by selective poly(ethylene glycol) precipitation of VLPs are defined with a view to demonstrating their potential in aiding the initial clarification stages of the purification sequence. The translation from laboratory scale to pilot scale operation is addressed, demonstrating the challenge of scale-up of solid-liquid separation stages for biological particle processing.
The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Torrington Place, London WC1E 7JE, U.K.
S
S
Tsoka
O C
OC
Ciniawskyj
O R
OR
Thomas
N J
NJ
Titchener-Hooker
M
M
Hoare
D001881:Borates / Q000737:chemistry; D011232:Chemical Precipitation; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D005427:Flocculation; D011092:Polyethylene Glycols / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000821:virology*; D014764:Viral Proteins / Q000737:chemistry / Q000302:isolation & purification*; D014771:Virion / Q000302:isolation & purification*
D001881:Borates; D014764:Viral Proteins; C018851:borax; D011092:Polyethylene Glycols