2.5.5 Fab expression pattern on dot blot
The FTN2 samples from microbioreactor cultivations were analyzed
initially in a high-throughput manner using dot blot to obtain a simple
yes-or-no-answer. For that purpose, 200 µL of cell suspension were
centrifuged in a 48-deep-well plate at 1200 rpm for 10 min. The
supernatant was removed and pelleted cells resuspended in 400 µL TE
buffer (pH 8). Resuspended cells were incubated for 1 hour at 60°C with
shaking, and then subsequently centrifuged as described above. The
supernatant (3 µL) was pipetted on a nitrocellulose membrane. Blocking,
incubation with antibodies, and membrane development were performed as
described below for Western blotting.