High-density perfusion culture of insect cells with a biosep ultrasonic filter.
The baculovirus/insect cell expression system has provided a vital tool to produce a high level of active proteins for many applications. We have developed a very high-density insect cell perfusion process with an ultrasonic filter as a cell retention device. The separation efficiency of the filter was studied under various operating conditions. A cell density of over 30 million cells/mL was achieved in a controlled perfusion bioreactor and cell viability remained greater than 90%. Sf9 cells from a high-density culture and a spinner culture were infected with two recombinant baculoviruses expressing genes for the production of human chitinase and monocyte-colony inhibition factor. The protein yield on a cell basis from infecting high-density Sf9 cells was the same as or higher than that from the spinner Sf9 culture. Virus production from the high-density culture was similar to that from the spinner culture. The results show that the ultrasonic filter did not affect insect cells' ability to support protein expression and virus production following infection with baculovirus. The potential applications of the high-density perfusion culture for large-scale protein expression from Sf9 cells are also highlighted.
Zhang
J
J
Human Genome Sciences, Inc., 9410 Key West Avenue, Rockville, Maryland 20850, USA. junli_zhang@hgsi.com
Collins
A
A
Chen
M
M
Knyazev
I
I
Gentz
R
R
D000818:Animals; D016367:Baculoviridae; D019149:Bioreactors; D002460:Cell Line; D002470:Cell Survival; D002688:Chitinases / Q000096:biosynthesis* / Q000235:genetics; D004867:Equipment Design; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D018411:Spodoptera; D014162:Transfection / Q000379:methods; D014462:Ultrafiltration / Q000295:instrumentation / Q000379:methods; D014465:Ultrasonics
D011506:Proteins; D011994:Recombinant Proteins; D002688:Chitinases