Page 9 of Attachment B 
The ventilation systems, exhaust gas filtration, waste products control, 
biological safety cabinets, barriers to unrestricted access, and all 
interior room surfaces in the P-3 and P-2 laboratories conformed to the 
NIH Guidelines. The NIH Guidelines for the Pl-LS level does not require 
specific ventilation measures, or a distinctly confined area. Character- 
istics of fermentors used by Eli Lilly for organisms containing recom- 
binant DNA molecules are: 
1. Inoculation, sample collection, nutrient addition, and transfer 
of culture fluids can be performed with closed system piping. 
All pipes and feeder tubes are connected to a fermentor at its 
top. 
2. Each fermentor has its own separate system of feeder lines, 
with the exception of electrical service, steam, and pH control 
(the latter two of which are double-check valved). 
3. Agitator shafts which penetrate the fermentor shell have double 
mechanical seals and are top mounted on the fermentor. 
k. The 150-liter fermentors are operated under slightly negative 
pressure, but the 2000-liter fermentors are operated at a slightly 
positive pressure (approximately 5 psi). 
5. Sterilization of fermentor exhaust Is accomplished by double 
f i 1 tration. 
6. Drainage pipes at the bottom of fermentors are sealed closed. 
7. Before initial use, the ability to sterilize an entire fermentor 
and its ancillary piping is verified by a test protocol employing 
thermocouples. 
Validation Procedures 
Validation of thorough sterilization of the fermentor, inoculation device, 
and exhaust gases is the responsibility of a validation team. A design 
engineer, two systems technicians, and the lead operator make up the team. 
Thermocouples and a recording system is utilized to assure that all points 
of the fermentor have been effectively sterilized. Each team member holds 
equal responsibility in giving approval for production start up of the 
fermentor. Thus, each must be confident that containment, sterilization, 
and inactivation procedures and systems are effective for size of the 
system, the specific host organism, and the recombinant DNA molecule. 
The validation team also reviews the work procedures and practices of 
the operator during the fermentation cycle. Recommendations and changes 
in procedure can then be made by the team toward the prevention of 
breaching the containment design (resulting in exposure). 
6 
[ 509 ] 
