1. 1 Douglas C. Lee, PhD Plasma Protein Therapeutics Association BPAC April 2011 Plasma Protein Therapies

2. 2 Plasma derived therapies; Manufacturing sites in USA Plasma derived therapies and Recombinant therapies; Manufacturing sites in USA, Austria, Belgium, Switzerland, and Italy Plasma derived therapies; Manufacturing site in Germany and in USA Plasma derived therapies; Manufacturing sites in Italy Plasma derived and Recombinant therapies; Manufacturing sites in USA, Switzerland, and Germany Plasma derived therapies; Manufacturing sites in Spain and USA Plasma derived therapies; Canadian Based (North America Regional Member Only) PPTA Members

3. Plasma Protein Therapies are special The starting material is human plasma Individual product classes (Immunoglobulin, clotting factors) are separated in a complex manufacturing process They are stable products with a defined shelf life of several years They are global, i.e. distributed throughout the world Plasma protein therapies are distinct from the labile blood components used for transfusion They are among the most highly regulated medicinal products 3

4. Relative risk From the donor public to the patient Finished product Virus inactivation / removal steps NAT testing Testing donations Inventory Hold Donor selection Donor population www.pptaglobal.org Standards and Certification / Pathogen Safety Systems

5. Overview of Pathogen Safety Systems  Step 5: Virus Inactivation Virus Removal Step 7: Packaging Control Step 1 Donor Screening, serological test on individual donations  Step 2 Donation Testing Nucleic Acid Technology (NAT), VMT  Step 3 Inventory Hold and Lookback  Step 4 Plasma Pool Testing In-process control Steps 6: Quality Assurance GMP Step 8: Post-Marketing Surveillance Steps 1 to 5: reduction of theoretical virus risk

6. Testing 1.5 Reduction > 9 Risk reduction (log 10 ) Selection 1 Pathogen Safety The Safety Tripod Reduction Virus inactivation/removal is the most effective safety measure and is designed to inactivate/remove a wide range of viruses  robustness  model viruses

7. PPTA Member Companies Testing Strategies

8. Overview Pathogen Safety Testing Paradigm Donation Testing Production Pool Testing Quality Control point Fractionation Viral Marker Tests on individual donations HBV surface antigen HIV-1 & HIV-2 antibody HCV antibody Viral Marker Tests HBV surface antigen HIV-1 & HIV-2 antibody Nucleic Acid Testing HBV HIV HCV NAT Minipool Testing NAT HBV HIV HCV Remove Positive Units

9. PPTA Member Companies Risk Reduction Strategies

10. 10 Virus Safety Viral Inactivation and Removal: Effective in eliminating HIV, HBV, HCV and other blood-borne infectious agents Validation: Provide evidence that selected steps of the manufacturing process effectively inactivate/remove viruses Provide indirect evidence that the manufacturing process will inactivate/ remove a wide range of viruses including (emerging) enveloped and non- enveloped viruses of diverse physico-chemical characteristics

11. Solvent/Detergent Pasteurization Dry heat Low pH Methods of Virus Inactivation and Removal Caprylate Precipitation Chromatography Nanofiltration Risk Reduction Method Selection is Dependent on the Virus’ Physical and/or Chemical Nature and Should Not Impact Product Attributes. For HBV, both its size and susceptibility to inactivation agents are readily considered when assessing a removal step.

12. 3.Measure total infectious virus in input and output fractions and calculate a log reduction value (LRV) Virus Clearance Experiments 1.Add virus to Input Load Input Fraction Output Fraction Scale-down model 2.Process spiked material using a bench scale model of the production step Example: Total Input = 10 5 viruses Total Output = 10 1 viruses Reduction = 10 5 /10 1 = 10 4 LRV = Log 10 10 4 = 4 Input virus (spiked into experiment) Residual virus (leftover after process step) Log Virus Reduction (LRV)

13. Page 13 The model virus concept, i.e. using a panel comprised of a wide range of physicochemically diverse viruses for the validation of virus reduction steps, with the goal to predict the behavior of a specific virus of interest, such as HBV. For HBV, a practical, relevant (specific) model virus is not available. Nevertheless, for some virus inactivation/removal steps Herpesviruses (PRV and others) have been used as unspecific model viruses since they share some structural characteristics (enveloped, DNA viruses) The Model Virus Concept

14. HBV: Physical and Chemical Characteristics Polymerase protein Core proteins -- HBcAg and HBeAg Lipid envelope HBV DNA Envelope glycoprotein (HBsAg) Family Hepadnaviridae Lipid enveloped virus Spherical, 40 to 48 nm diameter Relaxed circular, partially ds DNA genome, 3.0 to 3.3 kb Genome associated with the polymerase protein and an electron-dense nucleocapsid composed of the core antigens, HBcAg and HBeAg Low resistance to physicochemical agents solvent/detergent mixtures (TNBP/polysorbate 80) non-ionic detergents (Triton X-100) caprylate heat treatment

15. Example of a Panel of Model Viruses No simple in vitro assay system exists to directly model HBV inactivation/removal in scaledown studies, therefore a panel of model viruses is used that have physical and chemical attributes similar to HBV.

16. 16 Examples for Overall HBV Reduction Factor (log 10 ) Factor VIII concentrates Company Virus ABCDE HSV-1> 15.2> 9.3 PRV> 11.3> 9.3 BHV> 19.00 HIV-1 > 13.1 > 9.4 BVDV > 9.4 > 10.3 VSV> 10.9

17. Page 17 Factor IX concentrates Company Virus ABCDE PRV> 15.5> 7.3NA HIV> 11.7 > 12.2 BVDV> 15.5 Examples for Overall HBV Reduction Factor (log 10 )

18. 18 Intravenous Immunoglobulin Company Virus ABCDE PRV> 17.7 > 22.9* > 20.8* > 23.2> 12.2 > 9.3* > 16.7* HIV > 20.3 > 14.0 BVDV > 16.6 > 16.9 WNV> 13.8> 9.9 *Overall HBV reduction factor for different formulations Examples for Overall HBV Reduction Factor (log 10 )

19. 19 Albumin Company Virus ABCDE PRV > 12.00* > 14.00* > 14.1* > 12.2* > 11.4> 16.3> 12.9 HIV-1 > 10.5 > 17.8 BVDV> 11.2> 16.3 *Overall HBV reduction factor for different formulations Examples for Overall HBV Reduction Factor (log 10 )

20. Examples: Kinetics of Virus Inactivation Page 20 Inactivation by Caprylic acid in IGIVInactivation by Dry Heat in F VIII Inactivation by S/D in F VIII

21. Page 21 References for Kinetics of Virus Inactivation Dichtelmüller et al. Contribution to safety of immunoglobulin and albumin from virus partitioning and inactivation by cold ethanol fractionation: a data collection from Plasma Protein Therapeutics Association member companies: Transfusion 2011, in press Dichtelmüller et al. Robustness of solvent/detergent treatment of plasma derivatives: a data collection from Plasma Protein Therapeutics Association member companies. Transfusion 2009, 49:1931-1943

22. Other Plasma Derived Therapies Data for these products were not presented but pathogen safety related data can be found on the FDA website or the websites of the manufacurers: Factor XIII Subcutaneous/intramuscular immunoglobulins Specific immunoglobulins Antithrombin III Alpha1-Proteinase Inhibitor C1 Esterase Inhibitor Fibrinogen S/D plasma Page 22

23. 23 Summary HBV is a lipid enveloped virus with a low resistance to physicochemical agents that are commonly used in the manufacturing process of plasma protein therapies Current pathogen testing paradigms for source plasma incorporate orthogonal approaches that includes redundant assay methods (NAT, VMT) and test points within the process (donation, production pool) The virus removal/inactivation capacity for HBV in today‘s manufacturing process is in the order of 9 log 10 or greater

24. 24 Conclusions Pathogen safety measures introduced by regulatory agencies and PPTA member companies have significantly improved the safety of plasma derived products relative to pathogens causing chronic diseases, such as HBV. Since 1988 no transmission of HBV by a plasma derived medicinal product has been reported. Regulatory requirements applied to plasma protein therapies are among the most stringent for any medicinal product. PPTA member companies have also introduced additional industry standards for donor selection/screening and testing requirements in the manufacturing process. Today, plasma protein therapies have a excellent safety record with regards to known and emerging pathogens

25. 25 Back up slides