Adeno-Associated Virus Hybridization (Protocol 1516.104016 M,A», IncJ : 
This assay detects adeno-associated virus DNA present in the test article as determined by 
Southern hybridization using a 32 P labeled DNA probe. 
Parvovirus B-19 Hybridization (Protocol 1516. 104017 M.A.. Inc.): 
This protocol is designed to detect parvovirus B-19 DNA that may be present using Southern 
hybridization with a 32 P labeled DNA probe. 
Replication Competent-Adenovirus Assay (Protocol 1514.031002 MA. incJ : 
This assay is designed to detect the presence of human adenovirus in the test article. Indicator 
cell cultures are inoculated and observed periodically for CPE, a passage is performed for 
enhancement. 
IV.D.2.b Characterization of adenovirus seed Lot 
Each seed lot will be prepared, cryoperserved and aliquots evaluated according to the following 
criteria: 
•Mycoplasma (M.A., lnc.)-see above 
•Sterility (M.A., lnc.)-see above 
•In vitro virus (M.A., lnc.)-see above 
•Adeno-associated virus (M.A., lnc.)-see above 
•Parvovirus B-19 hybridization (M.A., lnc.)-see above 
•Electron microscopy (M.A., lnc.)-see above 
•Replication competent Ad5-please see description of assays in section IV.D.2.e 
•Presence of functional Ad.CB-CFTR-see description of assay in section IV.D.2.e 
IV.D.2.C 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. 
IV.D.2.d Characterization of purified lots of Ad.CB-CFTR for therapy 
Aliquots of each lot will be subjected to the following tests before use: 
•Sterility (M.A., lnc.)-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. 
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