SPADA Meeting Book

Appendix A: Example of a Culture Verification Statement 191 192 Culture Verification Statement for Francisella tularensis LVS test culture lot 2425-3243 193

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Summary

This culture verification statement documents the relatedness of Francisella tularensis LVS 195 test culture lot 2425-3243 (Manufacturer A; Boston, MA) to the sponsor-specified index culture 196 lot 9210-2349 (Manufacturer A). 197

198

Passage History

F. tularensis LVS test culture lot 2425-3243 was derived from lot 9210-2349 via an 199 intermediate culture (lot 8434-6286). The production and handling of the test and intermediate 200 cultures are summarized as follows. 201 Lot 2425-3243 was derived from lot 8434-6286 by Manufacturer A on 20 August 2019 202 according to the following procedure. A 10 µL aliquot of thawed lot 8434-6286 was spread onto 203 Sheep’s Blood Agar (Remel) and incubated at 35°C with 5% CO 2 for 48 h. The bacterial growth 204 was suspended to 1.0 McFarland unit in Tryptic Soy Broth (Remel) supplemented with 12.5% 205 glycerol. The resulting suspension was aliquoted into 1 mL cryotubes and stored at -80°C. 206 Lot 8434-6286 was derived from the index culture (lot 9210-2349) by Manufacturer A on 14 207 July 2016 using a method identical to the one described above. 208

209

Orthogonal Testing

Manufacturer A used whole genome sequencing along with Biolog and Vitek 2 GN 210 phenotypic assays to compare F. tularensis LVS test culture lot 2425-3243 with index culture lot 211 9210-2349. These results, which are reported in the attached Certificate of Analysis (1) and 212

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