Centrifugal recovery and dissolution of recombinant Gly-IGF-II inclusion-bodies: the impact of feedrate and re-centrifugation on protein yield.
The impact of centrifuge feedrate and multiple centrifuge passes on protein yield following recombinant Gly-Insulin-like Growth Factor II (Gly-IGF-II) inclusion-body dissolution has been investigated. Altering centrifuge feedrate did not significantly improve the overall protein yield following dissolution. Improved centrifuge recovery at a low feedrate was offset by poorer inclusion body paste purity. This reduced purity resulted in a significant loss of protein during inclusion-body dissolution due to proteolysis. Multiple centrifuge passes improved the inclusion-body paste purity. This resulted in a net improvement in the overall protein yield following dissolution. This work demonstrates that a strong interaction exists between centrifuge performance and inclusion-body dissolution for protease-sensitive products such as Gly-IGF-II.
Wong
H H
HH
Department of Chemical Engineering, University of Adelaide, Australia.
O'Neill
B K
BK
Middelberg
A P
AP
D019149:Bioreactors; D001709:Biotechnology; D002498:Centrifugation / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005069:Evaluation Studies as Topic; D005285:Fermentation; D006801:Humans; D002479:Inclusion Bodies / Q000737:chemistry; D007335:Insulin-Like Growth Factor II / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D011994:Recombinant Proteins; D007335:Insulin-Like Growth Factor II