TECHNICAL NOTE
4
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Screening for optimal growth medium
The components in the growth medium can boost cell growth during protein expression,
and thus are a critical factor to consider during optimization of protein expression for
a given target. To explore the effects of different growth media, E. coli cells expressing
His-tagged HRV3C were grown overnight at 20 °C in two different media, Terrific Broth
(TB) medium with 0.5 mM IPTG or ZYM 5052 auto-induction medium
1
. When the clarified
lysates were analyzed, no significant effect of the growth medium on yield was seen
(Figure 3A and B) by either method — SDS-PAGE analysis shows bands of similar intensity
for both media, and Andromeda traces show peaks of a similar height. Both growth
media tested yield a similar amount of total target protein as well as folded target protein.
Figure 3: Samples collected from cultures grown in two
different growth media were analyzed via SDS-PAGE
and Andromeda. A) SDS-PAGE of the lysate (L), clarified
lysate (C), and purified protein (P). The expected
molecular weight of the protein of interest is indicat-
ed by an arrow next to the standard. B) Andromeda
unfolding profiles of clarified lysate samples collected
from cultures grown in TB medium with 0.5 mM IPTG
(purple) and ZYM 5052 auto-induction medium (yellow).
Unfolding profiles represent the average of at least
three technical replicates. The shaded area indicates
the standard deviation amongst all technical replicates.
First
Derivative
Temperature [°C]
30 35 40 45 50 55 60
TB medium
ZYM 5052 medium
A
B
