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Table 1 Schedule for transient transfection of HEK 293-6E cells and subsequent feeding

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