In-situ gas analysis for fermentation BlueSens gas sensor - apz

Dr. Holger Mueller
In-situ gas analysis for fermentation
- or the easy way to control my process!
BlueSens gas sensor GmbH
• Founded in July 2001 by Dr. Udo Schmale and Dr. Holger Mueller
- up to 11 employees in 2009
- global network of distributors
- market leader for gas sensors in bioprocesses in Europe
• Main Focus:
Gas analysis to monitor bioprocesses
- Pharmaceutical industry
- Research and development
- Food industry
- regenerative energy (biogas)
• Philosophy:
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Measuring where something happens (no sampling)
Main Market: Pharmaceutical Industry
• Process optimization (R&D)
• controlled Scale up
• Quality control (yeast, yogurt etc.)
• Process control
• Start culture (shake flask) to determine the optimal time for inoculation
• Fermentation process (e.g. insulin production, cell culture, yeast production)
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Biogas Market
Laboratory scale yield tests
• CH4 quality
• Produced gas volume
Biogas plant
• CH4 control just in the near of the engine
• Total analysis of CO2, CH4, O2 and H2S
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Laboratory Scale
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Gas analysis for the Scale up: “normal scale up”
CO2 / O2 gas analysis
M
M
M
M
Shake flask
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Small size multi-fermenter
Gas analysis for the Scale up: “modern scale up”
• Online calculation of OUR, CER and RQ
• Increasing reproducibility
• Increasing productivity
• Greater understanding about your process
• Metabolic flux analysis and mass-balance calculations
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Gas analysis for the Scale up: “modern scale up”
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Principal of BCpreFerm
CO2
O2
CO
IR-CO2-Sensor
CO
GL 45 with PTFE
steril barrier (0,22 µm) with know difusion
rate
O
2
2
O 2-
2
A
ZrO2-O2-Sensor
O2
GL 45 with PTFE
steril barrier (0,22 µm)
O2
GL 45 with PTFE
steril barrier (0,22 µm)
CO2 CO2
O2
GL 14 for sampling
O2
Substrat +n O2
n CO2+ Produkt X
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Example 1
Optimisation of media with BCpreFerm
9
N2-source, 1 Litre flaks
260 rpm
H30(CO2) 0,5
H35(CO2) 0,75
H34(CO2) 1
H36(CO2) 2
8
CO2-Concentration [Vol.%]
7
Sampling
6
5
4
3
2
1
0
-1
00:00
12:00
24:00
36:00
48:00
60:00
Time [hh:mm]
Source: Bayer Healtcare AG, Wuppertal
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72:00
84:00
96:00
Example 2
OD-measurement versus CO2-measurement concerning produced biomass
0,030
produced CO2 [mol/l]
0,025
Shake flask
CO2 mol/l 50ml
CO2 mol/l 150ml
0,020
0,015
0,010
0,005
0,000
0
2
4
6
8
10
12
optical density
Source: Hexal Forschungs GmbH
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Example 3
Carbon balance in S. cerevisiae by online and offline analysis
Online data – BlueSens gas sensors
Jan Heyland
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Offline data - HPLC
Example 3
Physiological growth parameter and carbon balance
Spec. yield coefficient
[Cmol /Cmol]
Spec. rates
[mmol /gh]
Carbon balance could be exactly closed and is reproducible
⇒ good method for quantitative physiological studies
Jan Heyland
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Production scale
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Control of fermentation processes
“normal” gas analysis
M
air in
CO2 / O2
gas analysis
air out
gas cooler
p ro c e s s c o n tr o l
• a lot of maintenance
• delay of data
• difficult to control the process
Transport of the gas to the analyzer!
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Process control with in-situ gas sensors
“modern” gas analysis conform to PAT (process analytical technology)
air in
M
CO2
O2
air out
RS 232
4 -20 mA
processcontrol
• low maintenance
• online data
• easy to control the process
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Measuring where something happens!
Process control with in-situ gas sensors
...or for
tubes
for smale
pipes...
...for hugh
pipes...
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Advantages of BlueSens’ sensors
• small investment cost
• no sampling
• no gas treatment
• very stable and reliable measurements
• usable with every fermenter brand
• high redundancy through multiple sensors
• easy installation
• fits to every tube or pipe
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The size doesn’t
matter!
Process control with multiple sensors
Parallel Cultivation conform to PAT
air in
M
CO2
air out
O2
air out
M
• easy communication via Ethernet
CO2
air out
O2
air in
CO2
RJ45 (RS232)
RJ45 (RS232)
air in
O2
air out
RJ45 (RS232)
O2
air in
CO2
RJ45 (RS232)
• complete control loops
• different drivers available
air in
M
CO2
air out
O2
air out
RJ45 (RS232)
O2
air in
CO2
RJ45 (RS232)
BACCom12
RS232 / TCP/IP
FermVis
(Online OUR,
CER and RQ)
LabView
MFCS/Win
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Example 4
Measurement of the Oxygen uptake rate (OUR) and Carbon dioxide emission rate
(CER) of a yeast fermentation
Phase 2
Phase 1
CER [kmol/h]
OUR [kmol/h]
Phase 3
Ereignis 1
Phase 4
<- Zeitachse ->
Source: Schering AG, Engineo GmbH, BlueSens gas sensor GmbH
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Phase 5
Example 6
Discontinuous feeding of molasses based on an ethanol control
2,0
15
1,5
10
1,0
5
0,5
0
RQ
Concentration [Vol.%]
20
CO2 [Vol.%]
O2 [Vol.%]
RQ
0,0
25
30
35
Time [h]
www.ohly.com
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