VIA. Strategy and Standard Operating Procedures 
The overall strategy for the production of Ad5-CB-CFTR for therapy is 
summarized in Figure 24. A master cell bank (MCB) of 293 cells will be established 
that has been evaluated for performance, in terms of production of recombinant 
adenoviruses, and for the absence of other pathogenic contaminants. The MCB will 
be infected with our initial Ad5-CB-CFTR virus to generate a lysate. Virus will be 
purified and cryopreserved in aliquots. The lysate and purified seed lot will be 
subjected to safety testing as described in section VT.G. and VI.H. 
Production of Recombinant Adenovirus 
293 Cells 
1 
Establish a Master Cell Bank (MCB) 
Prep of virus 
, seed lot ' 
Infect\Wlth,A"d(CB)GETR £’ 
riMake pysatel, 
, 
Purify and' character Ize-- - ~ 
. virus 
Wt . 1 M i 11 fi i® 
Prep of virus 
for therapy 
Infect with seed lot 
Make lysate 
Purify and characterize 
virus 
Figure 24 
The MCB will be plated and infected with the virus seed lot. Lysates will be 
harvested from the infected cells and evaluated for sterility and mycoplasma testing. 
Virus will be purified from the lysate, cryopreserved, and lots will be evaluated as 
described in section VI.G. and VI.H. 
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Recombinant DNA Research, Volume 17 
