For all expression systems, i.e. E.coli, yeast, insect cells (baculovirus) and mammalian cells, a test phase preceeds the production phase. In the Test Phase, expression is performed under standard conditions to evaluate the productivity of the expression system and to determine the culture volume necessary for the production of the amount of protein required by the customer. In the Production Phase, expression is performed in agreed culture volumes according to the yield of the recombinant protein as evaluated in the test phase. For details please refer to table below.
| |
E. coli |
Yeast |
Insect (baculovirus) |
Mammalia |
| |
Test Phase: Expression under standard conditions to determine the culture volume necessary for the production of the required amount of protein. |
| Culture volume for test scale expression |
Transformation
200 ml |
Transformation
PCR check
30 ml |
Generation of
baculovirus
2 ml |
Transfection
Selection
30 ml |
Test affinity purification*
and detection |
Test affinity purification,
SDS PAGE and
Western blot |
Test affinity purification,
SDS PAGE and
Western blot |
SDS PAGE and Western
blot |
Test affinity purification,
SDS PAGE and
Western blot |
| |
Production Phase: Expression under standard conditions in culture volumes fixed according to the customer´s requirements. Purification with Strep- or 6xHistidine-tag, other tags on request. |
Expression and
purification scales |
1 L
2 L
5 L
10 L
up to 300 L |
100 ml
500 ml
1000 ml
5000 ml |
High titer
1.0 L
5.0 L
12.5 L |
1 L eq. 0.25 m2**
5 L eq. 1.00 m2**
** adherent cells |
| Affinity purification* |
with Strep- or
6xHistidine-tag |
with Strep- or
6xHistidine-tag |
with Strep- or
6xHistidine-tag |
with Strep- or
6xHistidine-tag |
