GE Healthcare

3 4 5 6 8 10 12 14

FDA ph 90% 70% DNA DNA Cytodex TM Baxter Biosciences 3

FDA EMEA cytodex 1 4400 cm 2 /g cytodex 1 25 g 17-0448-01 cytodex 1 100 g 17-0448-02 cytodex 1 500 g 17-0448-03 cytodex 2 3300 cm 2 /g cytodex 2 25 g 17-0484-01 cytodex 2 100 g 17-0484-02 cytodex 2 500 g 17-0484-03 cytodex 3 2700 cm 2 /g cytodex 3 10 g 17-0485-01 cytodex 3 100 g 17-0485-02 cytodex 3 500 g 17-0485-03 DNA 4

B A B C D E 1 CHO r-hbsag Bu- tyl-s Sepharose 6 Fast Flow r-hbsag r-hbsag 90% 2 1982 Butyl-S Sepharose 6 Fast Flow 1991 WHO 34 r-hbsag 2 90% r-hbsag 5% DEAE Sepharose Fast Flow 9% Sepharose 6 Fast Flow 800 CCS r-hbsag 97% 1 2 3 r-hbsag 1 Butyl-S Sepharose 6 Fast Flow XK50/20 130 ml 300 ml CCS 12 mg rhbsag 0.6 M ph 7.0 Butyl-S Sepharose 6 Fast Flow r-hbsag Sephadex G 25 DEAE Sepharose 6 Fast Flow DNA Sepharose 6 Fast Flow r-hbsag 1. CHO r-hbsag 2. Butyl-S Sepharose 6 Fast Flow r-hbsag 5

Butyl-S Sepharose 6 Fast Flow CHO r- HBsAg Bytyl-S Sepharose 6 Fast Flow 11-0026-34 ph Butyl-S Sepharose 6 Fast Flow 25 ml 17-0978-10 Butyl-S Sepharose 6 Fast Flow 200 ml 17-0978-02 Butyl-S Sepharose 6 Fast Flow 1 L 17-0978-03 Sephdex G-25 coarse 100 g 17-0044-01 Sephdex G-25 coarse 500 g 17-0044-02 Sephdex G-25 coarse 5 kg 17-0044-03 Butyl-S Sepharose 6 Fast Flow BioProcess TM DEAE sepharose Fast Flow 25 ml 17-0709-10 DEAE sepharose Fast Flow 500 ml 17-0709-01 DEAE sepharose Fast Flow 10 L 17-0709-05 Regulatory Support File RSF Butyl-S Sepharose 6 Fast Flow 5 L 17-0978-04 0.45 µm HCP 750 kda Sepharose 4 Fast Flow HCP 60 Q Sepharose XL virus licensed HCP DNA FDA EMEA 750 kda 3. 6

3 1 Vero Sepharose 4 Fast Flow 4 DNA Q Sepharose XL virus licensed DNA 5 Sepharose 4 Fast Flow 18-1020-52 Sepharose 6 Fast Flow Q sepharose XL virus licensed 18-1123-82 Sepharose 4 Fast Flow Sepharose 4 Fast Flow 1 L 10 L 17-0149-01 17-0149-05 Sepharose 6 Fast Flow 1 L 17-0159-01 Sepharose 6 Fast Flow 10 L 17-0159-05 Q Sepharose XL Virus Lic 300 ml 17-5437-01 Q Sepharose XL Virus Lic 1 L 17-5437-03 Q Sepharose XL Virus Lic 4. Sepharose 4 Fast Flow 5 L 17-5437-04 1. * <100 ng ** * n.d. = ** dose = 45 µg 5. Sepharose 4 Fast Flow Q Sepharose XL virus licensed 7

DNA DNA DNA GMP 100 kda DNA DNA RNA DNA cgmp Sepharose 6 Fast Flow PlamidSelect SOURCE 30Q 300 kda DNA RNA DNA DNA 6 7 1 Sepharose 6 Fast Flow RNA 2 PlamidSelect oc DNA DNA 3 SOURCE 30 Q 6. AKTApilot AKTApilot Sepharose 6 Fast Flow BPG TM 100 AKTApilot Sepharose 6 Fast Flow DNA PlamidSlect FineLINE TM 70 AKTApilot PlamidSelect DNA SOURCE 30Q FineLINE TM 35 7. 2 PlamidSelect DNA DNA RNA DNA oc = ccc = 8

2 cgmp 3 SOURCE 30Q and SOURCE 30S 18-1107-12 Plamidselect-Supercoiled Plasmid Puricication Starter Pack 18-1161-73 DNA 28-4057-63 2005-03-08-BJ cgmp Supercoiled Plasmid Purification Starter Pack Sepharose 6 Fast Flow 1 L 17-0159-01 Sepharose 6 Fast Flow 10 L 17-0159-05 10 mg DNA cgmp Plasmidselect Purification Pack 1 pack 17-6002-94 Plasmidselect 200 ml 17-5257-02 Plasmidselect 1 L 17-5257-03 DNA Plasmidselect Xtra starter Kit Plasmidselect Xtra 1 pack 200 ml 28-4052-68 28-4024-02 Plasmidselect Xtra 1 L 28-4024-03 SOURCE 30Q 50 ml 17-1275-01 SOURCE 30Q 200 ml 17-1275-02 SOURCE 30Q 1 L 17-1275-03 2. DNA % CGE* 95 DNA % AGE** 95 mg/ml 3.9 E.U./mg DNA 0.05 RNA µg/mg DNA 0.02 DNA µg/mg DNA 0.97 µg/ml < 51 * ** 1 3. * Sepharose 6 Fast Flow** 92 PlamidSlect % 72 SOURCE 30Q % 98 % 81 * A260 ** PicoGreen TM Molecular Probes pdna PlamidSlect-Supercoiled Plasmid Purification Starter Pack 10 mg DNA Sepharose 6 Fast Flow PlamidSlect SOURCE 30Q cgmp 9

Adv i ii iii iv v DNA 8 Q Sepharose XL virus licensed 0.65/0.2 µm 300 kda Q Sepharose XL virus licensed Ad5-CMV-NTR Q Sepharose XL virus licensed Ad5 Sepharose 4 Fast Flow 300 kda Ad5 8. Ad5 Cobra Biomanufacturing 9. Q Sepharose XL virus licensed Cobra Biomanufacturing 10

Data File Q sepharose XL 18-1123-82 9 Capto Q, S, DEAE 11-0025-76 AAV4 AAV5 Capto MMC 11-0035-45 11 Capto MMC Poster Capto Q Rapid adenovirus purification using Capto MMC Capto Q, S, DEAE Code No. Code No. Q Sepharose XL (virus licensed) 18-1178-07 Further reading (4) 300 kda Capto MMC AAV Capto Q 300 kda Purified recombinant AAV 4eGFP 10. A. Capto MMC AAV4 B. Capto Q AAV4 11. AAV4 11

12 FMD Frenkel BHK PEG Frenkel BHK 12 BHK

PEG 10 000 0.2 µm 0.45 µm Lumen E 20 LMH 3A 85 75 Lumen C E 40 LMH 500 KDa UniFlux membrane filteration system 18-1177-25 3-6 PEG 48 12. BHK 13

Fast Trak Custom Designed Media CDM Fast Trak CDM Fast Trak Education Regulatory Support Files Fast Trak www.gehealthcare.com/protein_purification Fast Trak Fast Trak Fast Trak 14

Manufacturing Process development Fast Trak Chromatography systems Chromatography columns Chromatography media Membrane systems Membrane devices Research Microcarrier matrices 15

1. Belew, M. et al. Purification of recombinant hepatitia B Surface antigen produced by transformed Chinese hamster ovary (CHO) cell line grown in culture Bioseparation 1, 397-408 (1991). 2. Scale up of a cost-effective and generic purification process for plasmid DNA. Life Science News Vol 16, 16-17, Code No 11-0003-48. 3. Scale-up of a method for highly pure plasmid DNA. Downstream 38. 20-21 (2004). Code No 11-0012-70. 4. Brument, N. et al. A versatile and scalable two-step ion-exchange chromatography process for purification of recombinant adenoassociated virus serotypes-2 and -5. Molecular Therapy 6. 678-686 (2002). 5. Herzer, S. et al. Key issues in the process development of purification procedures for viral vector and plasmid biopharmaceuticals. BioProcessing Journal 4, 67-72 (2005). 6. Pitarresi, T. M. et al. Purification of plasmid DNA on an anion exchange column without the use of RNase to process scale. Molecular Therapy 9, [Supplement 1] 53-54 (2004). 7. Herzer, S. Process design and scale-up of viral vector gene therapy products: New purification challenges. Cell & Gene Therapy 1, 43-48 (2004). 8. Diogo, M. M. et al Studies on the retention of plasmid DNA and Escherichia coli nucleic acids by hydrophobic interaction chromatography. Bioseparation. 10, 211-220 (2001) 9. Ferreira, G. N. Cabral, J. M. and Prazeres, D. M. Purification of supercoiled plasmid DNA using chromatographic processes. J. Mol. Recognit. 11, 250-251 (1998). 10. Diogo, M. M. et al. Production, purification and analysis of an expemental DNA vaccine against rabies. J. Gen. Med. 3, 577-584 (2001). 11. Jendrek, S. and Ekstrom, D. et al. Development of a production and purification method for type 5 adenovirus. BioProcessing Journal 4, 42-47 (2005). 12. Blanche, F. et al. An improved anion-exchange HPLC method for the detection and purification of adenoviral particles. Gene Ther. 7, 1055-1062 (2000). 13. Purification human influenza virus vaccine - towards a generic strategy. Downstream 37, 20-21 (2004) Code No 11-0008-46. http://www.gelifesciences.com.cn lifesciences@ge.com 8 8 33 12 (852) 2100 6314 (852) 2100 6338 1 (010) 5806 9689 (010) 6787 1162 100176 2401 (021) 5257 4650-67337 (021) 5208 1282 200336 1212 (020) 8363 3828-67961, 67956 (020) 8363 3291 510060 42 12 A-C (028) 8678 2581 (028) 8678 2582 610017 GE imagination at work 2007-09-26-BJ