1. 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
Colon Cancer
Macular Degeneration
Common in geriatric patients: Causes vision loss
2nd most deadly form of cancer
50,000 deaths a year
Year
U.S. Citizens Affected by Macular Degeneration
2017
9.1 million
2050
17.8 million
Customer Requirements
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
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 -
0.0103
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
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
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
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
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= %) $0
PBP
0.001
NPT (i=10%) 2
DBEP
2.001
NRR
517932%
DCFRR %
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