|
June 13
|
|
8:30 a.m. |
Welcome and introduction |
| 8:45-9 a.m. |
Workshop overview (Bentley) |
| 9-10:30 a.m. |
Lecture: Cell Physiology and Medium Requirements
(Payne) |
| 10:30-10:45 a.m. |
Break |
| 10:45-11:45 a.m. |
Lecture: Fermentors and Accessories,
Probes and Instrumentation (Bentley) |
| 11:45 a.m.-1 p.m. |
Lecture: Overview of Laboratory Experiments (Bentley)
| • |
high oxygen, low glucose, rich media |
| • |
low oxygen, low glucose, rich media |
| • |
low oxygen, high glucose, minimal
media |
| • |
high oxygen, high glucose, minimal
media |
|
| 1-1:30 p.m. |
Lunch Break |
| 1:30-2:30 p.m. |
Exercise 1: Familiarization with
fermentors (attendees assigned to each of the four types).
Examine fermentors, take vessel, etc., apart, reassemble,
make and record all needed measurements, add water, attach
tubing for air, acid/base and supplement addition. (Bentley,
Woodard, Marten, Payne) |
|
2:30-3:15 p.m.
|
Exercise 2 (begin): Prepare vessel
for medium addition; add medium, prepare vessel for autoclave
or in-place sterilization, sterilize. Cool down will be
handled by BSF personnel (staff).
|
| 3:15-3:30 p.m. |
Break |
| 3:30-4:15 p.m. |
Lecture: Recombinant E. coli
Protein Expression and Fermentation (Bentley) |
| 4:15-5:15 p.m. |
Exercise 2 (continue): Examine sterilized
fermentors in fermentation suite; regulate airflow and
agitation rate, see which impact dissolved oxygen tension.
(Bentley, Woodard, Staff) |
| 5:15 p.m. |
Adjourn
|
|
June 14
|
|
8-8:10 a.m. |
Meet in conference room for introduction to day two schedule. |
| 8:10-8:30 a.m. |
Exercise 3 (begin): Attach vessels
to fermentor; add other medium components; start gassing,
heating, etc.; check everything again for proper fit,
etc. Obtain inoculum in actively growing state, record
OD of fermentor. |
| 8:30-8:45 a.m. |
Exercise 3 (continue): Each group
inoculate fermentor, check everything; record information
needed; check again; take first sample, record OD. |
| 8:45-9 a.m. |
Exercise 4: Test YSI glucose analyzer, test
blanks and fermentor media; record in logbook. |
| 9-9:15 a.m. |
Break |
| 9:15-10:00 a.m. |
Lecture: Oxygen Consumption/
Transfer (Payne) |
| 10-10:30 a.m. |
Exercise 3 (continue): Take sample,
record OD, glucose levels. |
| 10:30-11:15 a.m. |
Case Study (Payne) |
| 11:15-11:45 a.m. |
Mini-lecture: Evaluation of run performance;
growth rate, doubling time, etc. (Bentley) |
| 11:45 a.m.-12:15 p.m. |
Exercise 3 (continue): Check fermentors;
record information; sample; start data graphing; ask questions,
visit other fermentors. |
| 12:15-1 p.m. |
Lunch break |
| 1-1:30 p.m. |
Exercise 3 (continue): Sample fermentors;
record data; graph data. |
| 1:30-2:30 p.m. |
Lecture: Scale-up of Fermentations:
Principles, Practice and Problems (Marten). |
| 2:30-3:00 p.m. |
Exercise 3 (continue): Check fermentors,
sample, record information, plot data; start yield and
growth rate calculations using glucose and OD data. |
| 3-3:30 p.m. |
Case Study (Marten) |
| 3:30-3:45 p.m. |
Break |
| 3:45-5:00 p.m. |
BSF demonstration of in situ sterilization,
cell separation and disruption,
questions and answers. |
| 5 p.m. |
Adjourn
|
