June
9 |
|
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
10 |
|
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
|
