From: High-titer expression of recombinant antibodies by transiently transfected HEK 293-6E cell cultures
Original transfection protocol | Improved standard transfection protocol | |||
---|---|---|---|---|
- 48 hrs | Cell seed at 5·105 mL-1 or | ↓ | ||
- 24 hrs | Cell seed at 1·106 mL-1 | - 24-2 hrs | Cell seed at 1-2·106 mL-1 | |
0 hrs | PEI-mediated transfection | 0 hrs | Cationicpolymer-mediated transfection | |
24-48 hrs | Tryptone feed (0.5% TN1) | |||
48 hrs | Harvest of intracellular proteins or | 48 hrs | Hydrolysates (0.5%) + volume doubling | |
72 hrs | Glucose feed (4.5 g L-1) | |||
96 hrs | Sodium valproate feed (3.75 mmol L-1) | |||
120 hrs | Harvest of extracellular proteins | |||
144 hrs | or | |||
168 hrs | or | |||
192 hrs | Harvest of extracellular proteins | |||
Antibody yields in the range of 130 to 150 mg L -1 | Antibody yields in the range of 500 to 800 mg L -1 |