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Table 3 Periplasmic extraction methods tested

From: Development of an automated platform for the optimal production of glycoconjugate vaccines expressed in Escherichia coli

Conditiona

t [h]

T [°C]

Protease inhibitor [Y/N]

Control

0.5

4

Y

1A

0.5

4

N

1B

1

4

Y

1C

1

4

N

1D

0.5

22

Y

2A

O/N

60

Y

2B

O/N

22

Y

2C

O/N

22

N

2D

O/N

30

Y

2E

O/N

30

N

2F

1

60

Y

2G

1

60

N

2H

1

45

Y

2I

1

45

N

2 J

1

30

Y

2 K

1

30

N

2L

1

22

Y

2 M

1

22

N

  1. aThe prefix for each condition represents Periplasmic Buffer 1 or 2. The subsequent letter (A–M) distinguishes the different conditions, changing either duration (t), temperature (T) or protease inhibitor tablet addition during periplasmic extraction. ‘O/N’ shows overnight incubation of ~ 16 h