Production of Monoclonal Antibodies using a Perfusion Tank

Production of Monoclonal Antibodies using a Perfusion Tank System Zero Flux Given: Brandon Blaesing, Brid Fillion, Thomas Gende, Sabrina Marnoto Introduction Process Flow Diagram Monoclonal Antibodies (MAbs) Seed Train • Represents 50% of the current biotech market Downstream Product Train Key TIC 14 Stream Number 1 Pressure; bar (gage) 25 • Common in geriatric patients: Causes vision loss • 2 nd most deadly form of cancer • 50, 000 deaths a year 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 Critical To Quality Variables Component Cells Media Mixture Glucose Variables Property Temperature Reactors 37 o. C Product Storage 4 o. C p. H Bioreactor p. H 7. 2 -7. 6 Dissolved O 2 >50% to promote optimal growth Viral Inactivation Stream Total Flow Rate (kg/batch) Temperature (C) Pressure (Bar) MW (kg/kgmol) 180. 156 1/5 2 3/6 8708 0. 0000015 228 25 37 25 1 1 1 4/8 325 25 1 7 9 10 11 18 8707 0. 78 8699 25 25 1. 2 - 0. 00000158 374 228 - 18 374 - 0 - 12 7. 84 25 1. 2 13 78. 4 25 1. 2 14 0. 7 25 1. 2 15 17. 0 25 1. 2 16 0. 1 25 1. 2 17 0. 7 25 1. 2 18 19/20 21/22 23/29 24 25 3. 87 0. 797 8. 76 88. 4 82 78. 3 25 37 37 37 25 25 1. 2 1. 2 374 - 0 - 3 - 0. 736 17. 00 0. 054 0. 68 - 0. 0103 0. 104 0 0 0. 003 0. 03 Air (w/o Oxygen) 28. 97 - - - 257 - - - - - 3. 055 - CO 2 44. 01 32 - - - 68 - - - - - 0. 812 - Water For Injection Product Citric Acid Buffer Solution Phosphate Buffer Saline Sodium Hydroxide 18 39. 997 8333 - - 0. 25 - - 17. 03 - - - - 53. 491 - - - - Ammonia Ammonium Chloride Occurs at p. H 3. 3 Protein A Impurities Ethanol Ammonium (NH 4+) 258. 06 46. 07 - 0. 02 8333 0. 75 8325 - 75 0. 00081 0. 01864 5. 942 E-05 0. 0007 - Single Use Reactors X X X ü - - - 337 - 34 - 49. 7 160. 1 - - 5. 47 37 3. 3 23 374 33 - 192 3. 055 5. 580 56. 24 5. 580 - - 36 156 - - 17. 2 16. 4 14. 95 - - - 1. 006 1. 84 18. 5 51 1. 48 - - 9. 57 41. 5 - - 0. 75 - 8. 25 - 83. 3 - - - - 0. 0003 0. 0029 0. 0294 8250 0. 23 - - - - - - 7500 750 0. 055 0. 18 - 36. 0 156 11. 8 51. 3 - 20. 5 0 0. 014 56 57/58 59 60 61 62 63/64 65 66 67/70 356 376 9580 647 2311 943 1319 86 1363 403 25 25 25 1. 2 1 1 1. 2 1. 0 1. 2 1. 0 - - - 2. 23 368 32. 7 - - - - 2. 2 368 32. 7 - - - - - - - - - - 323 - 379 - 346 - - - - - - - - 8333 - - - 3. 43 0. 0078 - - - - 68 69 71 72 73 74 75/76 77 78 81 82 83 84 85 86 2663 12243 24 1368 1753 13991 366 366 32 61 323 385 348 25 25 25 25 1. 0 1. 2 1. 2 - - - 0. 099 0. 32 2. 3 2. 2 374 370 368 33 33. 0 32. 7 55 1718 25 1. 2 78. 8330 8247 - 1677 349 8247 646 2258 930 1278 7 1328 379 2606. 32 10853 22. 4 1328 1707 12560 346 19. 0 18. 8 - 18. 6 0. 244 19 18. 4 0. 19 0. 43 0. 184 0. 61 18. 2 18. 0 - 41. 87 7. 29 - 34. 58 41. 87 - 930 930 - 53. 58 13. 74 - 39. 84 53. 58 0. 32 - 0. 646 0. 323 3 6. 8 3 4. 89 - 1. 95 4. 89 1. 95 - 1. 55 0. 17 1. 37 0. 17 1. 54 - Relief Valves • Spring relief valves • Bursting valves • Moderate: Temperature and level indicators on all reactors Profitability 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 Conclusions and Recommendations Conclusions 100 L Reactor 50/50 Cell Split Out Stream • Plant is a GO • Will be profitable within 3 years 0. 323 - - 1. 95 - 4. 89 32 - 0. 323 - - 4. 89 1. 95 - Reactors Storage Vessels Gas Movers 1% 0% 3% 0% 5% 6% Mixers • Selling price: $25 MM/kg Recommendations • Monitor media components • Increase titers to 8 -10 g/L • Ferment for 48 more hours NPV(i=0. 10) = $82. 4 B NPT(i=0. 10) =. 001 years $1, 000. 00 Separators Heat Exchangers i=10% for years 0 -2 $3, 000. 00 84% • $16. 2 Billion net profit after tax • 10% discount rate : $82. 4 Billion profit in 10 years Recirculated Stream - 56. 3 323 - 10% Discount Rate Total equipment cost: $13. 8 MM ($1, 000. 00) ($3, 000. 00) 0 1 2 i = 10% ($5, 000. 00) ($7, 000. 00) ($9, 000. 00) ($11, 000. 00) ($13, 000. 00) ($15, 000. 00) Feed Stream 346 17. 8 - Economic Analysis • Simplify: Citric acid & Na. OH buffer & regenerate • ½ cells inoculate the process trains A/B/C • ½ cells return to reactor - 371 210 - • Substitute: Sterilization of media with filter Perfusion Tank • 100 L perfusion reactor • Splits contents in tank - 37 38 39 40 41/42 43/46 44 45 47/79/80 48/49 50 51 52/53 54 49. 7 160 45. 6 197 879 8764 48 207 24. 1 8759 8671 8669 930 25 25 37 37 25 25 25 1. 2 1 1 1 1. 2 64. 9 61. 9 56. 2 • Minimize: Optimization of media components CIP/SIP ü ü ü X - - 34 35 36 7 7837 784 25 25 25 1. 2 - Inherent Safe Design Equipment Fouling Cost Safety Controllability Accountability Size - 30 31 32 33 243 4. 06 7. 42 74. 8 25 37 37 37 1. 2 1 1 1 Safety Novel Concepts Single Use 1. 04 4 0. 3 26 27 28 71 8621 210 25 25 25 1. 2 Cumulative Discount Cash Flow (USD) U. S. Citizens Affected by Macular Degeneration Valve sciences-practice-tut/e/antibody-class-differences Colon Cancer Year Temperature o. C https: //www. khanacademy. org/test-prep/mcat/biological-sciences-practice/biological- Macular Degeneration https: //www. macular. org/what-macular-degeneration Pump CWTR Utility Designation Symbol • Expected market growth: $125 billion by 2020 • Project based off Avastin™ (Bevacizumab) Control Systems DBEP(i=0. 1) =2. 001 Years End of Year (dated from project inititation) Acknowledgements We would like to acknowledge the chemical engineering department, Dr. Jeffrey Halpern, Future Dr. Levintov, Emily Poworoznek, Mihwa Palmieri from GE, and Greg Williams from Fisher Scientific 3