Automated Sampling Systems...# 2 • 25 employees in Massachusetts ... 0.420 mL/min. Gradient...
Transcript of Automated Sampling Systems...# 2 • 25 employees in Massachusetts ... 0.420 mL/min. Gradient...
Innovative Automation for Biotechnology # 1Innovative Automation for Biotechnology
Automated Sampling Systems
Cell Culture and Fermentation
Innovative Automation for Biotechnology # 2# 2
• 25 employees in Massachusetts• 8 representatives across the US• 13 distributor partnerships throughout Europe and Asia
Pacific
-
UK
BENELUXGermany
France
Sydney
KOREA
Thailand
ITALY
CHINA
Auckland
INDIA
Sweden, Norway, Denmark
PORTUGAL UAE
Indonesia, Singapore
JAPAN
Innovative Automation for Biotechnology # 3# 3
SanitaryReactor Barrier
Sample
Draw,
Prep
And
Delivery
Data
Analysis
Feedback
to
Controller
Optimization of
Fermentation or
Cell Culture
AutomatedClosedLoop
Solution
Phase 1
Phase 2
Phase 3
Innovative Automation for Biotechnology # 4# 4
Collaborations with Industry Leaders Providing Sample Draw, Prep and Analysis
HPLC
Nutrient Monitoring
Cell Viability
Fraction Collection
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Automated Sampling Automated Delivery Automated Analysis
Split DeliveryOne Sample – Four Analyses
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Fraction Collector
• The ARS-M delivers samples to sterile vials in the Fraction Collector for post run analysis. – Peltier Temperature Control– Maintains Temperature in a Range of 4°C to 37°C
• Optional Subzero Cooling with -40°C Chiller
Vial Size
Fraction Collector 60 Chilled Positions 15 ml
Fraction Collector 96 Chilled Positions 2 ml
Fraction Collector 24 Chilled Positions 50 ml
Fraction Collector Non-Refrigerated 50 ml
Autosampler 60 Position Chilled Tray 15 ml
Innovative Automation for Biotechnology # 7# 7
ManualSampling
Minimal Data Points
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Around-the-ClockOnline Sampling
All Day, All Night, All by Itself
Innovative Automation for Biotechnology # 9Innovative Automation for Biotechnology
ARS-M 840 Eight Inputs to 4 Instrument OutputsARS-M 440 Four Inputs to 4 Instrument OutputsARS-M 140 One Input to 4 Instrument Outputs
ARS-M SERIESAUTOMATED REACTOR SAMPLER
Innovative Automation for Biotechnology # 10# 10
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Volumes needed for analytical analysis
4 ml
4 ml
3 ml
6 ml
ARS
Cell Counter
HPLC
2.5L Working Vol
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Reactor Sampling VolumeGoal is to limit total volume removed to be <10% of initial working volume
100 150 200 250
1000
1500
2000
2500
Volume Available for Sampling, ml
Working Volume
10% Volume
Reactor Size Limitation on Sampling
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Biopharma
Animal Cell Culture
Ecoli_Yeast
Filamentous Fungi
Increasing Difficulty of Sampling
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DiptubeCell containing sampling
RIM ARSDiptube
Autoclave Sanitize in place, chemical
Glass or S.S. Bioreactor
Clamp/valve
Tubing sized to sampleviscosity
Connect by sanitary QC or tube welder
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Using dip tube-peristaltic loop for viscous samples
RIM ARS
Steam in place Sanitize in place, Chemical
Glass or S.S. Bioreactor
Clamp
tubing
Note: Peristaltic loop can be connected to top of head plate for Autoclaved Glass reactors. In line filtering available for cell free sampling.
Peristaltic Pump and loop
Connection by sterile QC or tube welder
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On line HPLC
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Groton-Agilent Online HPLC Assay System
• The Groton AAMK (Agilent Autosampler Modification Kit) turns the Agilent Autosamplerinto a Simple Fraction Collector.
• Once a fraction is collected in a vial or well plate the ARS-M starts the Agilent ChemStation and the 1200 technology is standard.
Fully Programmable, Fully Automatic24 Hours a Day, 7 Days a Week of Sampling
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ARS-M Hardware Upgrade Kitfor Agilent 1200 Autosamplers
3
4
1
6
2
5
Sample-Vial
Seat
Needle
Metering Device
Waste
Buffer Loop
Reactor pump input
Waste
6
3
4
1
2
5
Waste
ARSFLUSH
ALSMAIN
Internal plumbing of G1367D Hi-Performance Automated Liquid Sampler (Hi-P ALS)
Groton adds a valve and modifies the Agilent Autosampler internal valve to allow for Fraction Collection.
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ChemStation Integration• Set vial position where next fraction from
ARS-M will be collected.
• Set ChemStation methods for running samples, etc. within ARS-M commands.
• Start listening for commands from ARS.
• Stop listening to commands from ARS.
• Manually execute an ARS-M command.
When ChemStation is not listening for commands from the ARS-M, it can be used in regular analytical mode as a standalone instrument.
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On Line HPLC- Biofuel or Biopharma
min1 2 3 4 5 6 7
LU
0255075
100125150175200
0
LC: Agilent 1200 Series Rapid Resolution LCColumn: Agilent ZORBAX RRHT Eclipse Plus-C18, 2.1 x 50 mm, 1.8 µmDetector: Agilent 1200 Series Fluorescence DetectorSample: Amino acids, 50 pmol on column
Mobile phase A: 10mM Na2HPO4 – 10mM Na2B4O7, pH 8.2Mobile phase B: ACN:MeOH:H2O 45:45:10 by volumeFlow rate: 0.420 mL/minGradient timetable was used
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AUTOMATION FOR ETHANOL PRODUCTION
MKT-105 Rev A
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Biofuel System for On-line HPLC
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Biopharma Case StudyAutomation vs Manual Sampling
30 Liter B.Braun - 25 mm Ingold Side Port SIP Valves for Autosampling
Analysis Accuracy / Precision
Fraction Collector 2ml – 15 ml
Viable Cell Count ≤ 5%
Viability ≤ 2%
pH ≤ 0.035 Units
pO2 ≤ 7%
pCO2 ≤ 7%
Glucose, Lactate NH4 ≤ 7%
Amino Acids ≤ 10%
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Cedex Interface
0.0
5.0
10.0
15.0
20.0
25.0
30.0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
Elapsed Time (days)
Prod
uct C
once
ntra
tion
(mg/
L)
50.0
60.0
70.0
80.0
90.0
100.0
110.0
Cedex VCC Manual Cedex VCC Groton Cedex Via Manual Cedex Via
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PH Evaluation
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Glucose Evaluation
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Lactate Evaluation
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Future of Bioreactor Sampling
• Biopharm: Measurement of critical process parameters ( i.e. amino acids) along with product, ie. Glycosylated proteins. Feedback control
• Biofuel: Measurement of both fermentation and saccharification process. Enzyme monitoring