U.S. Citizens Affected by Macular Degeneration ZERO Production of Monoclonal Antibodies using a Perfusion Tank System Zero Flux Given: Brandon Blaesing, Brid Fillion, Thomas Gende, Sabrina Marnoto GIVEN Introduction Process Flow Diagram Monoclonal Antibodies (MAbs) Seed Train Product Train Downstream Key TIC Control Systems Pump 14 Stream Number 25 Temperature oC Represents 50% of the current biotech market Expected market growth: $125 billion by 2020 Project based off Avastin™ (Bevacizumab) 1 Pressure; bar (gage) CWTR Utility Designation Symbol Valve https://www.khanacademy.org/test-prep/mcat/biological-sciences-practice/biological-sciences-practice-tut/e/antibody-class-differences Colon Cancer Macular Degeneration Common in geriatric patients: Causes vision loss 2nd most deadly form of cancer 50,000 deaths a year https://www.macular.org/what-macular-degeneration Year U.S. Citizens Affected by Macular Degeneration 2017 9.1 million 2050 17.8 million Customer Requirements https://www.cancer.ca/en/cancer-information/cancer-type/colorectal/statistics/?region=on] 1,000 kg/year of MAbs 95% humanized sequence and 5% mouse sequence Chemically defined medium Purify MAbs to as pure as possible Material Balance   Stream 1/5 2 3/6 4/8 7 9 10 11 12 13 14 15 16 17 18 19/20 21/22 23/29 24 25 26 27 28 30 31 32 33 34 35 36 37 38 39 40 41/42 43/46 44 45 47/79/80 48/49 50 51 52/53 54 55 56 57/58 59 60 61 62 63/64 65 66 67/70 68 69 71 72 73 74 75/76 77 78 81 82 83 84 85 86 Total Flow Rate (kg/batch) 8708 0.0000015 228 325 8707 0.78 8699 7.84 78.4 0.7 17.0 0.1 3.87 0.797 8.76 88.4 78.3 8621 210 243 4.06 7.42 74.8 7837 784 49.7 160 45.6 197 879 8764 48 207 24.1 8759 8671 8669 930 1718 356 376 9580 647 2311 943 1319 1363 403 2663 12243 1368 1753 13991 366 323 385 348 Temperature (C) Pressure (Bar) 1 1.2 1.0 Component  MW (kg/kgmol) Cells - 0.00000158 0.0103 0.104 1.04 5.47 23 20.5 2.3 2.2 2.23 Media Mixture 374 3 4 371 337 370 368 Glucose 180.156 0.736 17.00 0.054 0.68 0.003 0.03 0.3 160.1 3.3 0.014 33.0 32.7 Air (w/o Oxygen) 28.97 257 3.055 64.9 61.9 56.2 192 5.580 56.24 156 36.0 CO2 44.01 1.006 1.84 18.5 11.8 51.3 O2 0.812 17.2 16.4 14.95 1.48 9.57 41.5 Water For Injection 8333 0.25 0.02 0.75 8325 7.5 75 0.00081 0.01864 5.942E-05 0.0007 8.25 83.3 8250 0.23 7500 750 0.055 0.18 833 3.43 8330 8247 1677 349 646 2258 1278 78.7 1328 379 2606.32 10853 22.4 1707 12560 346 56.3 Product 0.0078 19.0 18.8 18.6 0.244 19 18.4 0.19 0.43 0.184 0.61 18.2 18.0 17.8 Citric Acid Buffer Solution 258.06 41.87 7.29 34.58 Phosphate Buffer Saline Sodium Hydroxide 39.997 53.58 13.74 39.84 Ammonia 17.03 0.646 0.323 Ammonium Chloride 53.491 6.83 4.89 1.95 Protein A Impurities 1.55 0.17 1.37 1.54 Ethanol 46.07 Ammonium (NH4+) 0.0003 0.0029 0.0294 0.099 0.32 Critical To Quality Variables Variables Property Temperature Reactors 37oC Product Storage 4oC pH Bioreactor pH 7.2-7.6 Dissolved O2 >50% to promote optimal growth Viral Inactivation Occurs at pH 3.3 Novel Concepts Safety Economic Analysis Relief Valves Inherent Safe Design Profitability Total equipment cost: $13.8MM 10% Discount Rate Single Use Reactors Spring relief valves Bursting valves Minimize: Optimization of media components Substitute: Sterilization of media with filter Moderate: Temperature and level indicators on all reactors Simplify: Citric acid & NaOH buffer & regenerate Fixed Capital Investment $13.8 MM Working Capital $2 MM Total Capital $15.9 MM NPV (i=0%) $162.3 Billion NPV (i=10%) $82.4 Billion NPV (i=4014.22%) $0 PBP 0.001 NPT (i=10%) 2 DBEP 2.001 NRR 517932% DCFRR 4109.44% Salvage Value $1.7 MM Single Use CIP/SIP Equipment Fouling ü X Cost Safety Controllability Accountability Size Selling price: $25MM/kg $16.2 Billion net profit after tax 10% discount rate : $82.4 Billion profit in 10 years Perfusion Tank 100L perfusion reactor Splits contents in tank ½ cells inoculate the process trains A/B/C ½ cells return to reactor Feed Stream Conclusions and Recommendations Acknowledgements Conclusions Recommendations Recirculated Stream We would like to acknowledge the chemical engineering department, Dr. Jeffrey Halpern, Future Dr. Lev Levintov, Emily Poworoznek, Mihwa Palmieri from GE, and Greg Williams from Fisher Scientific Plant is a GO Will be profitable within 3 years Monitor media components Increase titers to 8-10 g/L Ferment for 48 more hours Out Stream 100 L Reactor 50/50 Cell Split