After the inoculation hose is connected to
the reactor and steamed for 20 minutes, the
expanded seed stock is pumped into the reactor
containing the media.
And now fermentation begins.
The operator takes zero hour readings and
monitors batch temperature, agitator RPMs,
dissolved oxygen levels, pH, vessel pressure,
optical density, air flow rate and glucose
concentrations.
When the targeted levels of glucose and optical
density are achieved, it's time to add IPTG
to the vessel to “turn on” the expression
of the Green Fluorescent Protein in the cells.
After about 5 hours, final readings are taken
and a sample is drawn to check the percentage
of cell solids.
We now refer to the product as broth…it
contains spent media and cells.
The process is complete when the glucose is
mostly consumed, and the batch has reached
the desired concentration.
The batch is then cooled down, pumped into
a broth tank...and labeled with the batch
number, volume, time and date.
The harvested broth will now move downstream
to Recovery where the cells will be ruptured
to free the GFP…And the protein will be
separated from the other broth components.
