Production of recombinant proteins in high‐density insect cell cultures

Open Access
Publisher Website
Google Scholar
Abstract
The effect of the growth phase of Spodoptera frugiperda (Sf9) cells on the production of recombinant proteins (β-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 β-galactosidase/106 cells and 1.7 μg glucocerebrosidase/106 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 × 106 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. © 1993 John Wiley & Sons, Inc.