1. PRODUCTION OF HIGHLY PURIFIED BOVINE TRANSFERRIN Group 5B Arjun Goel, Connie Marcelli, Lianne Kark, Sara Dang, Vivien Tai Faculty Advisor Prof C.A. Mims

2. Timetable

3. Agenda Mass Balances & Waste Treatment Key Unit Discussion Unit Specifications Current Challenges

4. Process Overview Crude Paste Filtration Chromatography Dry Transferrin Powder

5. Agenda Mass Balances & Waste Treatment Key Unit Discussion Unit Specifications Current Challenges

6. Mass Balance — Feeds Starting Paste 18 kg D.I. Water 571 L Celite 6 kg FeCl 3, NaHCO 3, Na-Citrate for iron Binding Diafiltration Buffers Wash for Column EDTA for Iron Chelating Process Distilled Water 160 L

7. Mass Balance — Products Water 740 L Celite 6 kg Inorganics FeCl 3, NaHCO 3, Na-Citrate, EDTA, NaOH Purified Apotransferrin 350g / Batch Process Trace Proteins Ethanol 4.5 kg SolidsLiquids Immunoglobins for recovery

8. Waste Treatment--Liquids Option 1 Option 2 Neutralize Acid/Base Addition To Sewer Waste Water Solid Waste Waste Water Process Reverse Osmosis Ion Exchange

9. Waste Treatment — Solids 1.Reverse Osmosis cartridges 2.Filter press cakes Disposal via contractor Mainly salts and celite trapped in filters

10. Agenda Mass Balances & Waste Treatment Key Unit Discussion Unit Specifications Current Challenges

11. Key Unit: R-101

12. Stirred Tank Cylindrical Conical Bottom Tank Material: SS316 In Cold Room 0-5 o C

13. Key Unit: R-101 Volume of solution in tank: 400 L Unit Selected: –Capacity: 757 L ( 200 Gal ) –Depth: 1.46 m –Diameter: 0.91m

14. Key Unit Accessories Level Indicator pH Monitor Temperature Indicator SS316 piping Valve Air Diaphragm Pump Mixer

15. Agenda Mass Balances & Waste Treatment Key Unit Discussion Unit Specifications Current Challenges

16. Filter Press: FP-101 12” in diameter Stainless Steel plates and frames Cellulose Pads Four 1” sludge frames 40 psi Cost: $6 703

17. Filter Press: FP-101 3 Applications: 1)4 Plates: removes 12 000 g paste @ 19 L/min 2) 2 Plates: removes 1 320 g denatured protein @ 9 L/min 3) 2 Plates: collects 3 600 g precipitated protein @ 9 L/min

18. Prefiltration SF-101 Before ultrafiltration Millipore low protein binding cartridge filter Filtration Area: 0.7m 2 80 psi Entrains particles > 0.5  m US$208 for 2

19. Ultrafiltration

20. Spiral Wound Filter UF-101 Concentration: 450 L to 90 L Polysulfone Flow: 100 L/hr Flux: 22 L/m 2 hr Pressure: 60 PSI Cost: $8 212

21. Spiral Wound Filter UF-102 Concentrate from 30L to 15L Regenerated cellulose Membrane Pressure: 60 PSI Flow: 10L/hr Flux: 11 L/m 2 hr Cost: $2 154

22. Hollow Fiber UF-201 Concentrate from 12 L to 6 L Polysulfone 25 PSI 6 L/hr 30 L/ m 2 hr Peristaltic Pump

23. DEAE Column

24. DEAE Sepharose Fast Flow Column: C-201 Automated Includes: The Column The Bioprocess Skid The computer DEAE Resin Total Cost: CND$211, 000

25. Water Distillation Column T-204 Rate: 190 L/24 hrs Storage capacity: 280 L Size: 2m x 1m x 0.6m Cost : US$ 3 945

26. Deionized Water Unit I-101 Flow range: 15 – 23 L/min Size: 1.5m x 0.6m x 0.4m Cost: US$ 4 142

27. Lyophilizer L-101 Similar unit used in plant Capacity: 8-12 L ~6 L of water to condense per batch

28. Future Directions Key unit development –P&ID –Design equations –Control Philosophy Economic Evaluation Plant Layout Appendices