SPADA Draft Documents

Appendix A: Example of a Culture Verification Statement

187

188

Summary

This Culture Verification Statement documents the relatedness of Francisella tularensis LVS 189 test culture lot 2425-3243 (Manufacturer A; Boston, MA) to the sponsor-specified index culture 190 lot 9210-2349 (Manufacturer A). 191

192

Passage History

F. tularensis LVS test culture lot 2425-3243 was derived from lot 9210-2349 via an 193 intermediate culture (lot 8434-6286). The production and handling of the test and intermediate 194 cultures is summarized as follows. 195 Lot 2425-3243 was derived from lot 8434-6286 by Manufacturer A on 20 August 2019 196 according to the following procedure. A 10 µL aliquot of lot 8434-6286 was spread onto Sheep’s 197 Blood Agar (Remel) and incubated at 35°C with 5% CO 2 for 48 h. The bacterial growth was 198 suspended to 1.0 McFarland unit in Tryptic Soy Broth (Remel) supplemented with 12.5% 199 glycerol. The resulting suspension was aliquoted into 1 mL cryotubes and stored at -80°C. 200 Lot 8434-6286 was derived from the index culture (lot 9210-2349) by Manufacturer A on 14 201 July 2016 using a method identical to the one described above. 202 204 phenotypic assays to compare F. tularensis LVS test culture lot 2425-3243 with index culture lot 205 9210-2349. These results, which are reported in the attached Certificate of Analysis (1) and 206 detailed on the manufacturer’s website (2) identified 5 single nucleotide polymorphisms in 207 noncoding regions of the bacterial genomes and generated identical genus and species calls on 208 Orthogonal Testing 203 Manufacturer A used whole genome sequencing along with Biolog and Vitek 2 GN

10