Beruflich Dokumente
Kultur Dokumente
STUDENT NUMBER
: 2012862664
GROUP
: EH2226A
EXPERIMENT
: LAB1
DATE PERFORMED
: 27 MARCH 2015
SEMESTER
:6
PROGRAMME / CODE
: EH222
SUBMITTED TO
No.
1
2
3
4
5
Title
Marks
Theory/Introduction
Methodology/Procedure
Process Flow Diagram (PFD)
Workbook/Reports
Results
TOTAL MARKS
100
Remarks:
Checked by:
Rechecked by:
----------------------------------(
)
Date:
Date:
1
TABLE OF CONTENTS
Theory/Introduction
1-2
Methodology/Procedure
3-4
Workbook/Reports
Results
Appendices
8-14
THEORY/INTRODUCTION
Monoclonal antibodies or MAB is a bioproduct that produced by an aerobic kinetic fermentation
in a batch fermentation tank. The tank is fed with media and biomass and also another air stream
applied to the fermentation to the fermenter. The table provided by the lecturer, gives the
stoichiometric coefficient of the overall reaction. Information given was extracted from an
experimental data. It is also stated that the Monod Kinetic equation = = 0.2 was
applied.
Reactants
Products
Component
Stoich. Coeff.
Component
Stoich. Coeff.
Media
100.00
Biomass
28.00
Oxygen
70.00
Carbon Dioxide
70.00
Impurity
10.00
MAB
2.00
Water
60.00
The batch fermentation tank can be operated in either Batch or semi-Continuous mode. The tank
equipped with default input, vent and default output. The default input is where the fed comes in
for example in this case, the media and oxygen flows in whereas default outputs is where the
product comes out such as MAB, impurity, biomass and water. There is another stream called
vent that allow the release of air if the pressure in the tank is over pressure and also if the tank is
under vacuum, it results in addition of air into the vessel so that the pressure inside the tank is
stable and equal to the atmospheric pressure. In this case, the carbon dioxide may produce and
release out tank through vent. (SuperPro, 1991)
Batch fermentation can be divided into few types such as kinetic, perfusion and stoichiometric.
During the first lab, it only involve kinetic fermentation which is a set of general equation provided
for the production rate was newly presented applying the kinetic theory on microbial cell growth.
Equations for product concentration in fermentation time courses were derived by developing
mathematically the general equation of production rate, and characteristic properties of
fermentation processes were clarified. Some examples of fermentations were analyzed kinetically
using the new kinetic theory. The calculated values of product, and cell concentrations were in
good agreement with the observed values. (T.Kono & T.Asai, 1969) Kinetic fermentation also
useful and potentially valuable for the improvement of batch process performance and essential
for continuous process design.
METHODOLOGY/PROCEDURE
1. SuperPro software was opened and new process file created.
2. Process model was started by clicking on the Task menu, and sat up the Set Mode of
Operation into the batch operation.
3. Pure components was then registered by using Pure Components item and the Register
Edit/View Properties was selected. The pure components that were registered are Media,
Oxygen, Nitrogen, Water, Biomass, Carbon Dioxide, Impurity, MAB.
4. Then, the Unit Procedures was clicked to find the Batch Vessel Procedure in a fermenter.
5. After the fermenter appeared on screen, the connect mode was switched on to draw the
streams in and out of the tank. The streams that need to be drawn are feed, water, air,
CO2 and broth.
6. Next, Operation Sequence set up opened when the fermenter diagram clicked twice and
then they was chose by referring to the question given by lecturer such as CHARGE-1
(Charge), CHARGE-2 (Charge), HEAT-1 (Batch Heating), FERMENT-1 (Batch Kinetic
Fermentation), COOL-1 (Batch Cooling), and TRANSFER-OUT-1 (Transfer Out).
7. In Operation Data menu, CHARGE-1 profile appeared after one right click on the
fermenter diagram and 10000 kg/batch of water was charged to the fermenter. This charge
operation was required a setup duration of 5 min and the processing time is calculated
based on 100 L/min.
8. In Operation Data menu, CHARGE-2 profile appeared after clicking the next sign and
duration of 20 min was setup and the processing time was calculated based on 10 kg/min.
The Feed stream was selected and the ingredient name or the component of Biomass
and Media was selected into the Composition menu. In the composition menu, a 500
kg/batch of a feed stream was setup at the Total flowrates, Set Mass Flow section. And
from the Flowrate (kg/batch) menu the Biomass was consisted of 50 kg and the rest is
Media was charged in.
9. In Operation Data menu, HEAT-1 profile appeared after clicking the next sign, Heating
Agent was change to steam. Fermenter was then heated to 37C by using steam at a
heating rate of 0.5C/min at a setup time of 5 min.
10. In Operation Data menu, FERMENT-1 profile appeared after clicking the next sign,
duration of fermentation set up for 24 hours. Chilled water was used as heat transfer agent
to maintain the fermenter at a final temperature of 37C. At Reactions Tab, the View/Edit
Kinetic Rate or symbol R was clicked and from the Kinetics for Fermentation menu the
Monod equation was selected and the Biomass was selected as the Rate Reference
Component and the B-Term and Media as the substrate.
11. In Operation Data menu, COOL-1 profile appeared after clicking the next sign, fermenter
was cooled to a final temperature of 5C by using Glycol at the cooling rate of 0.5C/min
at a setup time of 5 min.
12. In Operation Data menu, TRANSFER-OUT-1 profile appeared after clicking the next sign,
a process time of 5 hours was set up for further separation process.
13. Finally, all the Operation Data Menu completed and OK button was clicked.
14. After everything was done, Solve M&E Balance from the Task menu was clicked and the
process was check automatically if there is any errors available. If there is any, it will
appear as list at the bottom of page. Any errors listed need to be corrected until the
Perform M&E balanced ended successfully.
Water
carbon dioxide
37.00 C
1.01 bar
9863.83 kg/batch
8563338.77 L/batch
25.00 C
1.01 bar
10000.00 kg/batch
10053.24 L/batch
25.00 C
1.01 bar
500.00 kg/batch
500.01 L/batch
5.00 C
0.87 bar
10500.00 kg/batch
10476.82 L/batch
Broth
25.00 C
1.01 bar
9851.26 kg/batch
P-1 / FR-101
Fermentation
13249.57 L
CD = 33.81 h
Air
The diagram above is the process diagram of the fermentation process. The feed involves biomass, media,
air and water. Temperature inlet of the fed is 25C and pressure of 1.01 bar. Mass flowrate of water is
1000kg/batch while media is 500kg/batch and air has mass flowrate of 9851.26kg/batch. The product of
the kinetic fermentation are vent and broth. Carbon dioxide release out from the vessel with temperature
of 37C and flowrate of 9863.83/batch. Whereas, the broth flow out from fermenter with temperature of
5C and flowrate of 10500kg/batch.
WORKBOOK/REPORTS
Stream summary
Time Ref:
Batch
Type
Total Flow
Temperature
Pressure
Liq/Sol Vol
Flow
Total
Contents
Biomass
Carb. Dioxide
Impurity
MAB
Media
Nitrogen
Oxygen
Water
Time Ref:
Batch
Type
Total Flow
Air
Broth
kg
C
bar
L
Raw Material
9851.2560
25.00
1.013
0.0000
kg
kg
carbon
dioxide
Emission
9863.8279
37.00
1.013
0.0000
Water
10500.0000
5.00
0.867
10476.8198
Biomass and
Media
Raw Material
500.0000
25.00
1.013
500.0148
9851.2560
10500.0000
500.0000
9863.8279
10000.0000
0.0000
394.0502
0.0000
0.0000
0.0000
7092.9044
2364.3015
0.0000
Air
50.0000
0.0000
0.0000
0.0000
450.0000
0.0000
0.0000
10000.0000
Broth
10500.0000
0.0000
393.9283
0.0000
0.0000
0.0000
7102.6920
2367.2075
0.0000
carbon
dioxide
Emission
9863.8279
0.0000
0.0000
0.0000
0.0000
0.0000
0.0000
0.0000
10000.0000
Water
Raw Material
9851.2560
50.0000
0.0000
0.0000
0.0000
450.0000
0.0000
0.0000
0.0000
Biomass and
Media
Raw Material
500.0000
Raw Material
10000.0000
25.00
1.013
10053.2387
Raw Material
10000.0000
Gann Chart
day
h
1
8
16
2
24
32
40
day
48
Complete Recipe
P-1 in FR-101
CHARGE-1
CHARGE-2
HEAT -1
FERMENT -1
COOL-1
T RANSFER-OUT -1
CHARGE-1 (1.76 h)
CHARGE-2 (1.17 h)
HEAT-1 (0.48 h)
FERM ENT-1 (24.00 h)
COOL-1 (1.15 h)
TRANSFER-OUT-1 (5.25 h)
RESULTS
1. Amount of air to be supplied for the aerobic fermentation process is 9851.26 kg/batch.
2. The size of the fermenter that is necessary for this batch is 13249.57L.
3. The composition of vent are carbon dioxide, nitrogen and oxygen. CO 2 produced is 3.2440 %
and the flow rate is 9863.83 kg/batch.
4. The composition of broth are 1.6762% Biomass, 0.4286% Impurity, 97.8095% water and
0.0857% MAB and 10500kg MAB produced per batch.
5. The heating operation required is 0.48 hour and the cooling operation required is 1.15 hour.
6. The total time required to complete one batch is 33.81 hour.
7. The graph below shows the biomass attained stationary phase by providing the concentration
profiles for media, biomass and MAB.
Graph
45
40
35
Concentration
30
25
Concentration
Biomass
(g/L)
20
Concentration
MAB
(g/L)
15
Concentration
Media
(g/L)
10
5
0
0
-5
10
15
20
25
30
Time, h
APPENDICES
April 6, 2015
7,911.28h
33.81h
33.81h
234.00
MP = Undefined
10
Molar
Mass
Mass
(%)
(%)
(%) MP)
Main Section
(none)
(none)
kg/yr
kg/batch
2,340,000
10,000.000
11,700
50.000
105,300
450.000
92,208
394.050
1,659,740
7,092.904
Oxygen
553,247
2,364.301
TOTAL
4,762,194
20,351.256
kg/yr
kg/batch
2,340,000
10,000.000
11,700
50.000
105,300
450.000
92,208
394.050
1,659,740
7,092.904
Oxygen
553,247
2,364.301
TOTAL
4,762,194
20,351.256
Water
Biomass
Media
Carb. Dioxide
Nitrogen
kg/kg MP
kg/kg MP
11
% Total
kg/yr
kg/batch
P-1
100.00
2,340,000
10,000.000
TOTAL
100.00
2,340,000
10,000.000
% Total
kg/yr
kg/batch
P-1
100.00
11,700
50.000
TOTAL
100.00
11,700
50.000
% Total
kg/yr
kg/batch
P-1
100.00
105,300
450.000
TOTAL
100.00
105,300
450.000
% Total
kg/yr
kg/batch
P-1
100.00
92,208
394.050
TOTAL
100.00
92,208
394.050
% Total
kg/yr
kg/batch
P-1
100.00
1,659,740
7,092.904
TOTAL
100.00
1,659,740
7,092.904
% Total
kg/yr
kg/batch
P-1
100.00
553,247
2,364.301
TOTAL
100.00
553,247
2,364.301
kg/kg MP
Biomass
Procedure
kg/kg MP
Media
Procedure
kg/kg MP
Carb. Dioxide
Procedure
kg/kg MP
Nitrogen
Procedure
kg/kg MP
Oxygen
Procedure
kg/kg MP
12
Main Section
10,000.000
50.000
Media
450.000
Carb. Dioxide
394.050
Nitrogen
7,092.904
Oxygen
2,364.301
TOTAL
20,351.256
Main Section
2,340,000
11,700
105,300
92,208
1,659,740
Oxygen
553,247
TOTAL
4,762,194
3. STREAM DETAILS
Stream Name
Water
Biomass and
Media
Air
carbon dioxide
Source
INPUT
INPUT
INPUT
P-1
P-1
P-1
P-1
OUTPUT
0.00
0.00
0.00
0.00
25.00
25.00
25.00
37.00
1.01
1.01
1.01
1.01
994.70
999.97
1.20
1.15
0.00
0.00
13.06
46.08
0.00
0.00
1.14
4.02
1.00
1.00
0.24
0.24
Biomass
0.000
50.000
0.000
0.000
Carb. Dioxide
0.000
0.000
394.050
393.928
Media
0.000
450.000
0.000
0.000
Destination
Stream Properties
Activity (U/ml)
Temperature (C)
Pressure (bar)
Density (g/L)
13
Nitrogen
0.000
0.000
7,092.904
7,102.692
Oxygen
0.000
0.000
2,364.301
2,367.208
Water
10,000.000
0.000
0.000
0.000
TOTAL (kg/batch)
10,000.000
500.000
9,851.256
9,863.828
TOTAL (L/batch)
10,053.239
500.015
8,221,312.821
8,563,338.766
Stream Name
Source
Destination
Broth
P-1
OUTPUT
Stream Properties
Activity (U/ml)
0.00
Temperature (C)
5.00
Pressure (bar)
0.87
Density (g/L)
1,002.21
- 243.63
- 19.96
1.00
50.000
Media
450.000
Water
10,000.000
TOTAL (kg/batch)
10,500.000
TOTAL (L/batch)
10,476.820
14
INITIAL
INPUT
OUTPUT
FINAL
IN-OUT
Biomass
0.000
50.000
50.000
0.000
0.000
Carb. Dioxide
0.000
394.050
393.928
0.122
0.000
Media
0.000
450.000
450.000
0.000
0.000
Nitrogen
11.986
7,092.904
7,102.692
2.198
0.000
Oxygen
3.639
2,364.301
2,367.208
0.733
0.000
Water
0.000
10,000.000
10,000.000
0.000
0.000
TOTAL
15.624
20,351.256
20,363.828
3.052
0.000
15
5. EQUIPMENT CONTENTS
FR-101
Procedure
Operation
P-1
START
0.000
0.000
P-1
CHARGE-1 (Charge)
1.759
10,053.239
P-1
CHARGE-2 (Charge)
2.926
10,553.253
P-1
3.409
10,599.654
P-1
27.409
10,599.654
P-1
28.559
10,476.820
P-1
33.809
0.000
Time (in h)
Volume (in L)
16