VLH. 
Presence of functional Ad5-CB-CFTR-see description of assay in section 
VI.F. Characterization of cell lysate 
Each preparation of cell lysate will be pooled and aliquots will be evaluated 
for Mycoplasma and sterility (see above for descriptions) before virus is purified. 
Any lysate demonstrating contamination will be discarded. 
VI.G. Characterization of purified lots of Ad5-CB-CFTR for therapy 
Aliquots of each lot will be subjected to the following tests before use: 
Sterility (M.A., Inc.)-see above 
General Safety (Protocol 1514.509 M.A., Inc.): 
This assay is designed to detect extraneous toxic contaminants. Guinea 
pigs and mice are inoculated with test material and observed for overt 
signs of ill-health or unusual response. 
Limulus Amebocyte Lysate (LAL) (Protocol SPGT332070 M.A., Inc.): 
The purpose of this study is to detect and quantitatively determine the 
gram negative bacterial endotoxin level of a test article or extract using 
the Limulus Amebocyte Lysate (LAL) gel-clot method for testing. 
Replication Competent Ad5-please see description of assays in section 
VLH. 
Presence and quantity of functional Ad5-CB-CFTR-see description of 
assay in section 
VLH. 
VLH. Assays for functional CFTR virus and contaminating wild type Ad5 virus 
The final preparation of purified Ad5-CB-CFTR virus will be evaluated for 
the presence and concentration of virions capable of correcting the CF defect. The 
total particle number will be determined by measurement of the absorbance at 260 
nm. An aliquot of the virus preparation will be diluted serially and used to infect 
both 293 cells and the cell line CFPAC which was derived from a pancreatic 
adenocarcinoma of a patient with cystic fibrosis. The infected 293 cells will be 
evaluated for El transcomplementing virus using the previously descr ibed plaque 
forming assay. The infected CFPAC cells will be evaluated for CFTR protein 
expression by immunocytochemistry and correction of cAMP mediated chloride 
transport using SPQ assay. Descriptions of these assays are provided in section 
IV.A.2.B.(iii). 
Several experiments will be performed to evaluate the virus preparations for 
contamination with wild type adenoviruses. The presence of wild type adenoviruses 
in the 293 cell line will be evaluated by M.A., Inc. using a standard assay in which a 
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