generate the Master Cell Bank (completed November 30, 1992), and certification 
tests have been started. 
TABLE 1 
Process Testing 
Master Cell Bank Characterization of the cell bank 
Infect cells with adenovirus Adenovirus seed virus testing 
(adventitious contaminants) 
Cell Lysate Harvest Testing 
Purified Product*- Product Testing 
Ad5-CB-CFTR 
*Bulk Harvest and/or Final Fill Material 
VI.D. 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. 
VI.D.l. Sterility Test (Protocol 1514.510 MA, 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. 
VLD.2. Mycoplasma (Protocol 1514.102003 MA, Inc.): 
This protocol reflects the 1987 "Points to Consider" attachment and U.S. 
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. 
VI.D.3. Electron Microscopy (Protocol 1516.013 MA, Inc.): 
This protocol is designed to evaluate cell cultures for the presence of virus by 
examination of fixed cell pellets using transmission electron microscopy. 
VI.D.4. In Vitro Virus Assay (Protocol 1514.003 MA, 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 
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Recombinant DNA Research, Volume 17 
