Separation of recombinant human growth hormone from Escherichia coli cell pellet extract by capillary zone electrophoresis.
Free zone capillary electrophoresis (FZCE) was used to resolve recombinant human growth hormone (rhGH) and its variants from very crude mixtures of Escherichia coli (E. coli) cell paste extract. The methodology employs a phosphate deactivated fused-silica capillary and a 250 mM phosphate (pH 6.8), 1% (v/v) propylene glycol buffer with a high field strength of 600 V/cm. Resolution of rhGH and its variants from very crude mixtures did not change after 80 injections for the PSC capillaries. Bare silica and PVA coated capillaries had a more limited lifetime when injected with the same crude mixtures (10 to 30 injections). This FZC method provides a very powerful tool for assessing rhGH modification during the fermentation and isolation of rhGH that is not possible with other current analytical techniques.
McNerney
T M
TM
Department of Manufacturing Science, Genentech, Inc., South San Francisco, CA 94080, USA.
Watson
S K
SK
Sim
J H
JH
Bridenbaugh
R L
RL
D002457:Cell Extracts / Q000737:chemistry*; D002847:Chromatography, Agarose; D019075:Electrophoresis, Capillary / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics; D005285:Fermentation / Q000502:physiology; D019382:Human Growth Hormone / Q000031:analogs & derivatives / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D010710:Phosphates / Q000737:chemistry; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D012822:Silicon Dioxide / Q000737:chemistry
D002457:Cell Extracts; D010710:Phosphates; D011994:Recombinant Proteins; D019382:Human Growth Hormone; D012822:Silicon Dioxide