the purified product for endotoxin, microbial contaminants, extraneous toxins and infectious 
adenovirus completes the test battery. Each step will be carefully tested for the presence of 
wild type adenovirus. The majority of tests will be performed by Microbiological Associates 
Inc. (MAI), an independent laboratory that has provided contract research and safety assessment 
for the pharmaceutical industry. A contract has been established with MAI for this project. 
Characterization of the MCB should be completed in three months. 
T A BLE , 1 
Process 
Master Cell Bank 
Infect cells with adenovirus 
Cell Lysate 
Purified Product*- 
Ad.CB-CFTR 
*Bulk Harvest and/or Final Fill 
Testing 
Characterization of the cell bank 
Adenovirus seed virus testing 
(adventitious contaminants) 
Harvest Testing 
Product Testing 
Material 
IV.D.2.a Characterization of Master Cell Bank (MCB) 
The following tests will be performed on aliquots of the Master Cell Bank, these tests are based 
on the FDA's "points to consider" for use of mammalian cell lines. The MCB will be discarded if 
any test scores positive. 
Sterility Test (Protocol 1514.510 M.A.. Inc.) : 
This test exceeds 21 CFR 610.12 in utilizing three different broth media and one agar medium 
for detection of both aerobic and anaerobic bacterial as well as fungal contaminants. 
Mycoplasma. (P rotocol 15 1 4 .1Q20Q3 M .A., I nc.): 
This protocol reflects the 1987 "Points to Consider" attachment and US FDA expectations as to 
mycoplasma testing. The study includes direct inoculation of test article on two types of agar 
and in the semi-solid broth medium. Broth inoculated with test article is subcultured at three 
intervals and incubated aerobically and anaerobically. In addition to the agar and broth 
cultivation, direct inoculation of the test article on Vero indicator cells and staining with 
Hoechst stain permits microscopic examination for the presence of mycoplasma. 
Electron Microscopy (Pro tocol 1516.013 M.A.. Inc.) : 
This protocol is designed to evaluate cell cultures for the presence of virus by examination of 
fixed cell pellets using transmission electron microscopy. 
In Vitro Virus Assay (Protocol 1514.003 M.A.. Inc.) : 
A broad spectrum of viral contaminants are detected using an In Vitro Assay with sensitive 
indicator cells. Indicator cells, including the test article cells, MRC-5 cell line, and Vero cells. 
Test procedures include direct inoculation of test article as well as blind passage of supernatant 
fluids at 14 days. Indicator cells are observed for cytopathic effect for the entire 28 day period. 
Hemagglutination and hemadsobtion are performed twice during the assay. 
Recombinant DNA Research, Volume 16 
[829] 
