22nd VH Yeast Conference
Berlin, April 27 – 28, 2009
New Raising Power Measurement Devices
for
Advanced Tasks
Helmut Trautmann
abiotec AG
advanced bioprocess technologies
CH-5445 Eggenwil
How all began . . .
Custumer-Driven Development for
Centre de Recherche Nestlé (CRN) in Lausanne:
Low Temperature Inactive (LTI) Yeast Project
asks for a sensitive measuring device to detect
gassing ativities of 100- to 1000-fold below
standard yeast dough conditions
Systems on the market did not fulfill these criteria!
We: A Spin-off Company from ETH-Zürich were already in collaboration
with Nestlé in the topic of
Advanced Bioprocess Technology
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
„Quality Control by Process Control“
Presented at Hannover Trade Fair
and at Biotechnica
Our concept from 1994 . . .
. . . becomes 10 years later a major demand of US-FDA‘s PAT-Initiative !
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Process Analytical Technology
in „State of Routine“
Sterile sampling
from bioprocess
Industrial
Sampling
System
(ISS)
Sample transport
Sample processing
sample storage
in cooler
Online
Process
Analyser
(OPA)
Analysis
(HPLC, GC etc.)
Transfer of
validated results
Roche Vitamins / DSM:
Production Plant with
6 Bioreactors, each 140 m3
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Monitoring of Product, Byproduct and Cell Density
On-line Monitoring of Industrial Fedbatch Process
100
80
Product
By-Product
Product / By-Product
Concentration
(arbitrary units)
60
80
OD (660 nm)
70
50
60
50
40
40
30
30
20
20
10
Optical Density (AU, 660 nm)
90
70
10
0
0
0
10
20
30
40
50
60
Time (h)
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
User Requirements (selection)
• Lower detection limit at about 1 ml gas per day
• Suitable system-volume for samples of about 30g dough
• Duration of experiments up to 3 weeks
• Ambient temperature range from 0°C to 40°C
• User-friendly concept
• Modular extendable system
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Selection of Measuring Principle
1. Open Systems
1.1 Massflow Sensors
not sensitive enough
1.2 „Classical“ Displacement principles
Automated evaluation of measuring requires additional sensors,
complex handling
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Selection of Measuring Principle
2. Closed Systems
Measurement of Pressure-Differences by
2.1 Absolute Pressure Sensors
2.2 Relative Pressure Sensors
reference measurement required
Disadvantage: Management of overpressure and security aspects
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Realised System
Gasvolume-Monitor „Niesler“
(Name according to the Initiators Dr. Peter Niederberger and Dr. Christof Gysler from CRN)
Recording of absolute pressure over time,
calculation of generated gas volumes according to
Ideal Gas Law
p*V=n*R*T
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Gasvolume-Monitor „Niesler“
System Overview
Measuring Range: >4 Magnitudes
Lower Limit: 1 ml per day
Upper Limit: 3 L per hour
Resolution: 0.05 ml
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Gasvolume-Monitor Niesler
Windows-based Operation-Software
• Automated and manual interactions with measuring units
• Units can be freely combined in experimental sets
• Multiple sets can be registered simultaneously
• Results are presented directly via online view on the screen
• Excel-readable files are generated for subsequent user-specific operations
• Generation of standardised reports in QA routine measurements is possible
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Data from Dr. Christof Gysler, Nestec Lausanne:
Activity = f (Temperature)
Niesler® gas-monitor:
¾ gassing activity increases with
increasing temperature
ml CO2 / 35 g dough (160 mg dry yeast)
Baker’s yeast in model dough
50
10°C
40
8°C
30
6°C
20
4°C
10
2.4 ul CO2 / g *h
2°C
0
0
-10
10
20
30
hrs
40
50
60
70
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Data from Dr. Christof Gysler, Nestec Lausanne:
Activity = f (Recipe)
Niesler® gas-monitor:
e.g.: NaCl concentration
40
ml CO2 /35g
30
1.0%
1.2%
20
1.4%
10
1.6%
0
0
12
24
36
48
-10
hrs (at 8°C)
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Data from Dr. Christof Gysler, Nestec Lausanne:
Activity = f (Yeast concentration)
Initial activity (5-35 hrs) vs. [conc.]
Cumulative CO2 development
60
1.2
0.1%
activity (ml /35g dough/h)
activity (ml /35g dough
50
0.2%
40
0.4%
0.6%
30
0.8%
20
10
1
0.8
0.6
0.4
0.2
0
0
20
40
60
80
0
0
-10
Storage time (hrs,)
0.2
0.4
0.6
0.8
1
Dry yeast concentration (%)
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Data from Dr. Christof Gysler, Nestec Lausanne:
Activity = f (Time)
¾At low temperatures and low yeast conc. the gassing activity is linear
« normal » dry Yeast / LTI Yeast
300
200
mlCO2 / 35g dough
8°C
150
+ 65%
10°C
250
200
150
100
17 ul CO2 / g *h
50
100
+ 13%
50
0
0
0
10
20
30
hrs
40
50
60
70
0
10
20
30
hrs
40
50
60
70
Temperature-Profile of LTI-Yeast is not according Arrhenius
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Niesler: New Edition 2009
ROBUST AND PRECISE
• Fabricated completely in stainless steel
• Water protected measuring head
• User-friendly and efficient Tri-Clamp fastener
• „Plug and Work“-Layout
• Improved concept of components => improved gas-tightness
• Extended Security-Concept for Overpressure (Software, Firmware, Hardware)
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Niesler 2009: Reproducibility
Gassing activity from 100g model dough at 30°C,
measured in 3 different Units
7
6
ml CO2 / g dough
rel. StdDev ≤ 1%
5
4
3
2
Unit 1
Unit 2
Unit 3
1
0
0
1
2
3
4
hrs
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Comparison of Niesler with SJA Fermentograph
Experiments at VH-Berlin
by Dipl. Biotechnol. Claus Decker
were just started . . .
3.50
6.00
3.00
5.00
ml CO2 / g Teig
CO2 [ml/g Teig]
2.50
2.00
1.50
4.00
3.00
2.00
1.00
H 11623 Niesser 50g direkt
0.50
1.00
H11642 std Niesser 100g
H 11623 SJA
H11642 SJA
0.00
0.00
0.00
0
0.5
1
1.5
2
2.5
0.50
1.00
1.50
2.00
2.50
Zeit [h]
Zeit [h]
Spec. Gassing Activity Rates vs. Dough Size
2.50
20
15
2.00
5
Endabweichung [%]
0
40
50
60
70
80
90
100
110
-5
-10
-15
-20
120
130
140
150
160
ml CO2 / g * h
Endabweichung [%]
10
1.50
1.00
0.50
-25
-30
Teigeinwaage [g]
0.00
0
50
100
150
200
250
300
350
Dough Size (g)
Exemplary first results: Gassing activity as a function of sample size
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009
Niesler 2009:
Future Perspectives:
Development of new Evaluation and Reporting Tools
for Industrial Routine Applications
Development of other Vessel Formats
adapted to Customer-specific Work-Flows
. . . to be continued . . .
What are your specific needs ?
New Raising Power Measurement Devices for Advanced Tasks – 22nd VH Yeast Conference in Berlin, April 27-28, 2009