Continuous production and recovery of recombinant Ca~(++) binding receptor from HEK 293 cells using perfusion through a pached bed bioreactor

摘要

The extracellular domain of human parathyroid Ca~(++) receptor was needed for structural and clinical studies. The desired protein was produced by immobilizing the transformed HEK 293 cells in a packed bed configuration using a 2.2 liter bioreactor equipped with a vertical mixing impeller assembly and an internal basket. The production was done in two stages: a propagation stage followed by a production stage. In the first stage, which lasted approximately 140 hours, the bed was perfused with serum-containing medium, allowing the cells to grow at a constant growth rate. When the total cell population reached approximately 3 x 10~(10), the second stage was begun. Serum-free medium replaced the serum containing medium, and the perfusion process was continued for an additional 400 hours. During this time, the medium was pumped through the packed bed at a rate of 4-6 liters per day, keeping the residual glucose concentration at 1 gl~(-1).