1. IND Review Chart (for submission of IND to CDER)

2. NDA Review Chart (for submission to CDER)

3. A Warning Letter

4. Picture of a Clean Room

5. Diagram of the Flow Pattern of HEPA-Filtered Air Through a Typical Clean Room Incoming air supply HEPA filter Items of clean room equipment Exhaust unit Ceiling

6. Clean Room Classification How clean rooms are classified: Two standards are used to classify clean rooms. The first, Federal Standard 209E, is used domestically and the newer standard is TC 209 from the International Standards Organization (ISO). Both standards classify a clean room by the number of particles found in the laboratory’s air. If a laboratory is a class 10 clean room by the Federal Standards, there are less than 10 particles per cubic foot. The following chart relates FS to ISO classifications and recommended action levels of microbiological quality, i.e. the maximum levels in the clean room air that microorganisms should be allowed to reach:

7. Federal Standard (particles/ft 3 ) a ISOParticles/m 3 Microbiological Action Levels (cfu/m 3 ) b 1<1>0 24 1335>0 104352>0 10053,5201c1c 1,000635,2007 10,0007352,00010 100,00083,520,000100 a Particles are of size 0.5 μm; the standard varies for particles of different sizes. b cfu means colony forming units. c Samples from Class 100 (ISO 5) environments should normally yield no microbiological contaminants.

8. Generalized Clean Room Design Clean room Transfer lock door Entry of raw material Entry of personnel (changing room) Bench Transfer lock Exit of product Exit of personnel (changing room)

9. Photo showing some clean room attire

10. A Tri-Clamp Connection

11. Diaphragm Valves

12. Piping in Biomanufacturing

13. A Peristaltic Pump Principle of operation Rotor Flexible Tubing Fluid

14. Side Cross-Section of Centrifugal Pump

15. General Components of Centrifugal Pump

16. Rotary Vane Positive Displacement Pump

17. On/Off valve Steam (in) Water (in) Steam (out) Water (out) Diagram of a typical jacketed processing vessel Such vessels are usually made from high grade stainless steel By opening or closing the appropriate valves, steam or cold water can be circulated through the jacket In this way the vessels contents can be heated or cooled, as appropriate In addition, passage of steam through the jacket of the empty vessel will effectively sanitize its internal surfaces

18. The range of CIP agents most often used to clean/sanitize chromatographic columns 0.5-2.0 M NaCl Non-ionic detergents (0.1-1.0%) NaOH (0.1-1.0 M) Acetic acid (20-50%) Ethanol (~20%) EDTA (~1.0 mM) Protease solution Dilute buffer Most CIP protocols would make use of two or more of these agents, allowing them to percolate sequentially through the column at a slow flow rate. Contact time can range from several minutes to overnight. Sodium hydroxide (NaOH) is particularly effective at removing most contaminant types.

19. Sterilization-in- Place Module

20. Bulk Freeze-Thawing of Purified Protein Formulation Process (Addition of Necessary Buffers, Purified (Thawed) Protein, and other Ingredients) Preparation & Posidyne- Filtration of Required Buffers Filtration of Formulated Material through a 0.22 micron Filter Filling of Vials Schematic of Formulation-Fill Unit Operation Sequence (The unit operation train will be different for a sustained-delivery formulation or a formulation that requires the use of a delivery device)