control loops for glucose feed in cell culture and ... · pdf filecontrol loops for glucose...

Control Loops for Glucose Feed in Cell Culture and Microbial Fermentation

Bioprocesses Based on Online Measurement

Henry Weichert, PAT & Automation

IFPAC, January 2014

turning science into solutions

Migration from Process Development to Manufacturing

Develop Design Space Utilize Design Space

Data Information Process

Understanding Demo of

Understanding Process Control

DoE MVA

SCADA System

Sensors & Analytical Systems

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Cells as Platforms

Cells

Microorganisms Insect cells Animal cells

Bacteria Sf 9 Hybridoma

E.coli Sf 21 BHK

Yeast CHO

Pichia pastoris NS0

Saccaromyces cerevisiae

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Most relevant industrial biopharmaceutical processes

Methods (traditional Hybridoma)

Monoclonal antibodies

Recombinant proteins (Insulin)

Vaccines

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•developed by G.Köhler and C.Milstein in 1975

•Nobelprize 1984

Why Online Glucose | Lactate Measurement

Glucose

One of the key nutrients Basis of energy pathways

Global Trends Lactate

One of the key metabolites Influences pH | Indicator for cultivation conditions

Typical concentrations (at starting point)

Cell culture Glc 4-10 g/l Lac 0-5g/l Doubling time ~24h

Microbial culture Glc 20-60 g/l Lac 0g/l Doubling time ~ minutes

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Solution

Why Online Glucose | Lactate Measurement (2)

Problem

• Issues during manual sampling

• Time consuming, on weekends, during nights

• Risk of contamination

• Automated connection (sampling and communication)

• Direct measurement which represents cell culture status

• Sampling possibility for all setups (small volumes, SU and MU)

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Consistent Sensor Utilization along Scale-Up Example: Glucose-Lactate Measurement

R&D PD Clinical/Production




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BioPAT®Trace: Online Monitoring & Control of Glucose and Lactate

Glucose | Lactate measurement in real-time

Fully automated, self calibrating

Modes: Filtration, Dialysis

Plug & Play

Reusable system

SU sensor & fluidics set ready to use

Automated glucose feed control with

MFCS win + Recipe Module

Compact device

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BioPAT®Trace: Online Monitoring & Control of Glucose and Lactate

- Glucose | Lactate measurement in real-time

- SU sensor and fluidics ready to use

- Automated glucose feed control with MFCS win + Recipe Module

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Typical Setup of BioPAT®Trace

Calibration liquids

Transportation buffer

Analyzer Probe

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Online Sampling for Reusable Bioreactors – Filtration Probes

Advantages Robust polypropylene

membrane

Very fast response time

(up to 60 measurements / hour)

Various adapters

Variable length

Easy handling

But: Volume reduction

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Online Sampling for Reusable Bioreactors – Dialysis Probes

Advantages No volume reduction

Fast response time

(up to 30 measurements / hour)

Independent of media composition and viscosity

Various lengths and adapters

Diffusion membrane

Acceptor current

unloaded loaded

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Comparison to existing reference analysis

Measurement during Batch (CHO cells, measurement every 6h)

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Comparison to existing reference analysis

Measurement during Fed Batch (CHO cells measurement every 6h)

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Long Time stability over 45 day culture

Lo

Long time culture

of a CHO cell line

for 45 days.

Data provided by

University Bielefeld,

Prof.Gudermann

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BioPAT® MFCS/win: Sophisticated Process Control and Automation Consistent Process Data Base along Product Lifecycle

R&D PD Clinical/Production

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Process Automation & Control with Recipes

Recipe Editor Sequential Recipe Operations

Sequential & Parallel Phases

Review & Approve

￭ The necessary set of information that uniquely defines the production requirements for a specific product or operational procedure. [ANSI/ISA-88.01]

￭ Typical objective is to achieve process robustness and reproducibility through reliable automation to reduce risk of errors and free up operator time.

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Advantages of automated control setups

Recipes and PID Contoller

• Automated setup of feeding points

• Endpoint determination / end of culture / no glucose

• Automated calculations of uptake rates / per cell or per volume

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BioPAT®Trace – Integrated Glc | Lac Measurement and Process Control

Global Trends BioPAT®Trace

Parallel measurement of Glucose and Lactate

Dialysis or Filtration Mode Compact design for flexible

integration

DCU – Local Control Unit

Direct display of g/L Glucose | g/L Lactate

Quick and simple process monitoring

BioPAT®MFCS/win + Recipe Module

Recipe for Process Automation Automatic event based control of

cultivations: Adding feeds Aeration control

Process and yield optimization

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Setup PID Controller in BioPAT®MFCS/win

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Status in the Group Display

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Example of a 2 Point Glucose Controller in MFCS/win (S88)

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Basic Setup of the 2 Point Controller

Process Parameter BioPAT®Trace

Glc (g/l) Lac (g/l)

Standard 1 0,5 0,24

Standard 2 10,08 5,01

Measurement period 2 min

Glc-concentration (feed) 100g/l

Pump rate (100%) 10ml/min

Setup of controller in BioPAT®MFCS

Glucose_low 38% means 3,8 g/l

Glucose_high 40% means 4,0 g/l

Setpoint pump 10% means 1 ml/min

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Example: Glucose control in a CHO culture

CHO culture running over 12 days with cell dilutions in between

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BioPAT®Trace | Customer Projects – Lonza Hopkinton, MA

BioPAT trace - comparison to YSI

for more detailed info:

Lonza Hopkinton

Sohan Patel

[email protected]

Direct 774-759-8780

Configuration: Sartorius 40L Biostat C | BioPAT Trace with 90mm Dialysis probe (Ingold port adaptation)

Process details: Undisclosed (please contact Sohan Patel for specific details)

Primary Objective: Automate feed start based on glucose depletion –

currently labor intensive manual measure and start

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mailto:[email protected]

Conclusion

Develop Design Space Monitor Design Space




DoE MVA

SCADA System / MFCS

Sensors & Analytical Systems

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Thank you for your interest

www.sartorius-stedim.com/PAT

control loops for glucose feed in cell culture and ... · pdf filecontrol loops for glucose...

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