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Use and Availability:
The AAV2 reference material (RM) ATCC VR-1616 is available from ATCC at a nominal cost. Each vial contains 0.5 mL of AAV2 RM at 3.28 x 10E10 vg/mL, or 4.37 x 10E9 IU/mL with TCID50 analysis. The pTR-UF-11 vector plasmid ATCC MBA-331 used to produce the reference material is also available from ATCC.  However, the HEK-293 production cells are not available for sale, and you should plan to use your own cell bank to develop and test your internal reference material.  Other relevant materials available from ATCC include the HeLa RC32 cell line (ATCC CRL-2972) and the adenovirus type 5 helper virus (ATCC VR-1516) that are used to conduct the infectious titer assay.
Sterility Testing:
Prior to freezing, the filtered formulated bulk was sampled (5 mL) and tested for bioburden by AppTec (now WuXiAppTec, Inc.). It was found negative for aerobes, fungi, spores, and obligate anaerobes, with an assay sensitivity of <5 CFU/sample. The filtered formulated bulk was tested for endotoxin and found negative, with an assay limit of detection of 0.06 EU/mL.
The vialed rAAV2 RSS (ATCC VR-1616 lot 58051221) was tested for sterility and endotoxin at the Indiana University Vector Production Facility according to GLP guidelines. No aerobes, anaerobes or fungi were detected following direct inoculation onto thioglycollate broth, trypticase soy broth and Sabouraud dextrose agar and incubated for 14 days at the appropriate temperature. Negative and positive controls (Bacillus subtilis, Candida albicans, Bacteroides vulgatus) were included in the test. Absence of endotoxin was verified using the limulus amebocyte lysate gel clotting assay. Test samples were assayed in duplicate. Negative, positive and spiked controls were included. The sensitivity of the assay was 0.06 EU/mL.
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AAV RM Project Participants: