Biotechnology Research and Education Program
June 8  
 
8:30 a.m.
 
Welcome and introduction
8:45-9:00 a.m. Workshop Overview (Bentley)
9:00-10:30 a.m. Lecture: Cell Physiology and Medium Requirements (Payne)
10:30-10:45 a.m. Break
10:45-11:45 a.m. Lecture: Bioreactor Performance Criteria – Lag Phase, Growth Rate & Yield (Bentley)
11:45 a.m.-12:00 p.m.

Lecture: Overview of Laboratory Experiments (Bentley)

IPTG induced, chemically defined media, 10g/L glucose, no pH control, oxygen controlled
Autoinduced, complex media, no glucose, pH controlled, oxygen controlled
Arabinose induced, complex media, 5g/L glucose, no pH control, no oxygen control
12:00-1:00 p.m. Lunch Break
1:00-2:15 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. (Woodard, staff)

2:15-2:30 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).
2:30–3:15 p.m. Oxygen Consumption and Transfer (Payne)
3:15-3:30 p.m. Break
3:30-4:30 p.m. Lecture:  Recombinant E. coli and Protein Expression (Bentley)
4:30-5:15 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 Adjourn
 
   
June 9
 
8:00-8:10 a.m.
 
Meet in conference room for review of today’s 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.
8:45-9:00 a.m. Exercise 4: Test YSI glucose analyzer, test blanks and fermentor media; record in logbook.
9:00-9:15 a.m. Break
9:15-9:45 a.m. Mini-Lecture: Evaluation of run performance; growth rate, doubling time, etc. (Bentley)
9:45-10:30 a.m. Case Study:  Process Development for Enzyme Production (Payne)
10:30-11:00 a.m. Exercise 3 (continue): Take a sample, record OD, glucose levels.
11:00-11:45 a.m. Lecture: Interpreting Fermentation Data (Marten)
11:45 a.m.-12:15 p.m. Exercise 3 (continue): Check fermentors; record information; sample; start data graphing; ask questions, visit other fermentors.
12:15-1:00 p.m. Lunch break
1:00-1:30 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)
2:30-3:20 p.m. Exercise 3 (continue):  Check fermentors, sample, record information, plot data; start yield and growth rate calculations using glucose and OD data.
3:30-3:45 p.m. Break
3:45-4:15 p.m. Which group had the highest growth rate? Yield?
4:15–5:00 p.m.. BSF demonstration of in-situ sterilization, cell separation and disruption, questions and answers.
5:00 p.m. Adjourn

 

 

 

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