70 Life Technologies
™
| Clone Screening
Clone Screening
2-5 days
2-5 days
5 days
2-5 days
14 days
2-5 days
Transfer remaining volume of each clone to be scaled (e.g., top 100) up to
a well of a 24-well plate containing 0.3-0.5 mL of pre-warmed medium
Single-cell
protein-producing clones
in 96-well plate
Transfer 0.5 mL of each clone to a well of a 6-well plate containing
1.5-2 mL pre-warmed medium
Passage each clone in 6-well plate (static or shaking) (e.g., 1 mL
of each clone into 2 mL of pre-warmed medium)
SECONDARY SCREEN: Seed each clone to a well of a 6-well plate (static or shaking)
at 3 x 10
5
cells/ml in 3 mL of medium; sample for protein production on day 5
Passage top 30 clones at 2 x 10
5
-3 x 10
5
cells/mL,
10-30 mL, in SF125
Proceed to optimize protein production using different culture conditions, or
continue the scale-up procedure to meet your bioproduction needs
TERTIARY SCREEN: Seed top 30 clones at 3 x 10
5
cells/mL, 30 mL, in SF125,
shake at 150 rpm; perform 14-day simple fed-batch with glucose-only feeding
to identify top 10 clones; maintain cell trains
Figure 16. Clone scale up from 96-well plates to shake flasks.
