Production of recombinant proteins in high-density insect cell cultures

The effect of the growth phase of Spodoptera frugiperda (Sf9) cells on the production of recombinant proteins (beta-galactosidase and glucocerebrosidase) was investigated. Cells infected with the recombinant Autographa californica nuclear polyhedrosis virus at the late exponential and stationary pha...

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Bibliographic Details
Published in:Biotechnology and bioengineering 1993-06, Vol.42 (2), p.235-239
Main Authors: Reuveny, S, Kim, Y.J, Kemp, C.W, Shiloach, J
Format: Article
Language:English
Subjects:
ADN RECOMBINADO
ADN RECOMBINE
Animal cells
BACULOVIRIDAE
baculovirus
BETA GALACTOSIDASA
BETA GALACTOSIDASE
Biological and medical sciences
BIOREACTEUR
BIORREACTORES
Biotechnology
CULTIVO DE CELULAS
CULTURE DE CELLULE
Eukaryotic cell cultures
EXPRESION GENICA
EXPRESSION DES GENES
Fundamental and applied biological sciences. Psychology
GLICOSIDASAS
GLYCOSIDASE
high density culture
insect cells
Lepidoptera
MEDIO DE CULTIVO
Methods. Procedures. Technologies
MILIEU DE CULTURE
Miscellaneous
Noctuidae
PATHOGENESE
PATOGENESIS
recombinant protein production
SINTESIS DE PROTEINAS
SPODOPTERA FRUGIPERDA
SYNTHESE PROTEIQUE
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Summary:The effect of the growth phase of Spodoptera frugiperda (Sf9) cells on the production of recombinant proteins (beta-galactosidase and glucocerebrosidase) was investigated. Cells infected with the recombinant Autographa californica nuclear polyhedrosis virus at the late exponential and stationary phases yielded low quantities of expressed protein. Highest enzyme yields were obtained using Sf9 cells from the early exponential phase (0.9 mg beta-galactosidase/10(6) cells and 1.7 micrograms glucocerebrosidase/10(6) cells). Infection of resuspension of cells collected from various phases of growth in fresh medium resulted in 75% restoration of maximal expression levels. This finding suggested either nutrient limitation or waste product accumulation as the cause of the decrease in productivity at the latter phases of growth. Further experiments revealed that the highest productivity levels could be obtained with cultures of Sf9 cells grown in a fermentor to a cell concentration of 4 X 10(6) mL-1. The medium needed to be replaced prior to infection with the recombinant virus and supplemented with a mixture of glucose, L-glutamine, and yeastolate ultrafiltrate
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.260420211