Chapter 3 Fermentation Production Process Scale Up

School of Health Sciences &
Technology
Department of Biotechnology
CUBTM 219: Fermentation Biotechnology
Chapter 3: Fermentation Production Process Scale-up

(Lecture 4)
By
Dr. R. Kamusoko (DPhil, MSc, BSc)

Intended learning Outcomes
• Demonstrate understanding on the processes and

procedures for fermentation process scale-up
• Design a process scale-up for fermentation

production
Contents
• Scope of fermentation process scale-up
• Scale-up studies
• Rules followed during scale-up
• Plan for process scale-up
• Cell banking strategy for large scale manufacturing
• Seed train scale-up strategy
• Production fermentation scale-up
• Linear scale-up parameters

Scope of Fermentation Process Scale-Up
• Microbial fermentation plays a critical role in many industrial

applications
• Fermentation processes are utilised to produce a final product,

convert substrates, catalyse reactions, remediate
environmentally toxic materials or make mass biological
materials
• Process scale-up enables a fermentation achieved in research

and development to operate at commercially-viable scale
• Successful scale-up involves many aspects of preparation and

planning
Purpose of Scale-Up
• To ensure that fermentation at large-scale is techno-

economically viable
• Allows investigation of a product and process on small- and

intermediate-scale before large amount of money is invested on
large-scale
• To get the same fermentation efficiency as obtained in small-

scale fermenters at the most economical values
• Failing of process at industrial-scale causes million dollars loss
• Scaling-up gives investors more confidence on chances of

success against economic disaster
Scale-Up Studies
• Studies carried out at lab- or pilot-scale to yield data that

could be used to extrapolate and build large-scale fermenters
• The aim is to build industrial size fermenters capable or close

to producing fermentation products as efficient as those
produced in small-scale fermenters
• A process developed in a lab (e.g. 0.5 – 10 L fermenters)

must be translated to a full- scale manufacturing process
(e.g. 20 000 – 2 000 000 L fermenters) with scale factors
ranging from thousands to millions
cont..
• Scale-up of large-scale processes is done in 2 stages:
1. Pilot-plant (100-10 000 L fermenters)
− to translate lab-scale process into a realistic scale-down

version of the manufacturing process
2. Demonstration plant (demo-scale) (10 000 to 100

000L fermenters)
− to minimise the risk of a large capital investment in the

full-scale manufacturing plant by further validating the
process, supply chain and market demand
Rules to be Followed During
Scale-Up
• Few rules are followed when doing scale-up studies:
1. Similarity in the geometry and configuration of

fermenters used in scaling-up
2. A minimum of three or four stages of increment in the

scaling-up of the volume of fermentation studies
3. Each jump in scale should be by a magnitude or power

increase and not an increase of a few litre capacity
− Slight increase in the working volume will not yield

significant data for scale up-operation
Steps in Scale-Up Studies
 Define product economics based on projected market size

and competitive selling and provide guidance for allowable
manufacturing costs
 Conduct laboratory studies and scale-up planning at the

same time
 Conduct preliminary larger than lab studies with equipment to

be used to aid in plant design
cont..
 Design and construct a pilot plant including provisions for

process and environment controls, cleaning and sanitization
systems, packaging and waste handling system and
meeting regulatory agency requirements
 Evaluate pilot plant results (product and process) including

product economics to make any corrections and a decision
on whether or not to proceed with a full-scale plant
development
Uses of a Pilot Plant
• A pilot plant is used for:
1. Evaluating the results of lab studies, and making product

and process corrections and improvements
2. Producing small quantities of product for sensory,

chemical, microbiological evaluations, limited market
testing or furnishing samples to potential customers,
shelf-life and storage stability studies
3. Determining possible profitable by-products or waste

stream requiring treatment before discharge
cont..
4. Providing data that can be used in making a decision on

whether or not to proceed to full-scale operation, and in
case of a positive decision, designing and constructing a
full-size plant or modifying an existing plant
5. Reduce the risk associated with construction of large

process plants
 Computer simulations and semi-empirical methods are

used to determine the limitations of a pilot-scale
 Mathematical models are then tested in physical pilot

plant
Modeling Methods to Determine the
Limitations of a Pilot Scale
Various modeling methods are used including:
 Chemical similitude studies
 Mathematical modeling
 Finite elemental analysis
 Computational fluid dynamics

Cont..
 Chemical similitude studies:
− Computer aided modeling and simulations in the

development and integrated optimisation of the industrial
process
− e.g. Advanced Simulation Library
 Mathematical modeling:
− The process of using various mathematical structures -

graphs, equations, diagrams, scatter plots, tree diagrams,
and so forth to represent real world situations
Cont..
 Computational fluid dynamics:
− The use of applied mathematics, physics and computational

software to solve and analyze problems that involve fluid flows
− e.g. Sim scale, SciLab
 Finite elemental analysis:
− A computerized method for predicting how a product reacts to real-

world forces, vibration, heat, fluid flow, and other physical effects
− Shows whether a product will break, worn out, or work the way it
was designed
− e.g. Autodesk
Cont..
• Mathematical models yield information about:
− Finalized mass and energy balance
− Optimized system design and capacity
− Equipment requirements
− System limitations
• After data has been collected from operation of a pilot plant,

a larger production-scale facility may be built
Plan Process Scale-Up
• The first step is to define goals, targets, facility and timeline
for completion
• Common goals and targets include the following:
− Annual outputs of the products based on analysis of market
demand and manufacturing capacity
− Productivity targets based on process and facility capability
− Yield per lot based on process capability, facility scale and
operational capability
− Process cycle time based on throughput, process and
operational constraints
− Cost targets based on process capability, product price,
market demand and facility operating cost
Research in Scale-Up Process
• The exercise in scaling up involves several programmed
research that has to be established so as to predict the final
behaviour of the final large scale production fermenter. These
include:
1. Inoculum development – cell banking strategy
2. Sterilization to establish the correct sterilization cycle at

larger loads
3. Environmental parameters such as nutrient availability,

pH, temperature, DO, dissolved carbon dioxide
4. Shear conditions and foam production

Cell Banking Strategy for Large-Scale
Manufacturing
• A commercial fermentation process requires a stable cell bank

that ensures consistent and predictable production in a long
term
• A common cell bank strategy for large-scale manufacturing is to

maintain a master cell bank (MCB) from which a working cell
bank (WCB) is generated for routine production
• The strategy allows original source of genetically stable MCB to

be preserved for long periods of time
• Cell banks are stored in -70°C liquid nitrogen or at -80 °C freezer

Cont..
• The cell bank quality and stability need to be tested and monitored.
General tests of cell bank include:
1. Cell recovery upon freeze and thaw: growth test
2. Contamination or purity tests: colony morphology, genetic

discrimination tests, phenotypic tests, etc
3. Real-time stability tests: time points for growth, genetic stability

and product expression stability
4. Product qualification: use of small-production model to check the

cell bank’s suitability for production
5. A comparability study of the MCB and WCB recommended to

ensure WCB’s performance
Seed Train Scale-Up Strategy
• The purpose of this strategy is to propagate cells to a desired

mass for inoculation into the production bioreactor
• Traditional seed train strategy includes thawing a vial and

inoculating into a shaker flask for a certain number of stages
with increasing flask size and may include stainless steel reactor
• Guiding principle is to minimize the number of stages for the

entire seed train, maintain robust growth and prevent negative
impact of extended generations to productivity and product
quality
Cont..
• Typical studies of seed scale-up strategy may include:
1. Vial thaw conditions:
− include temperature and time between thaw and inoculation
2. Inoculum ratio:
− minimum inoculum ratio needs to be defined at every step
− ratio of 0.1 to 5% is commonly used for microbes
3. Seed train media:
− optimized to support growth rather than product expression
− use a simple recipe for seed train media
4. Genetic stability and production stability during seed train

process
A typical microbial seed train:
Linear Scale-Up Parameters
• One of the outcomes of a process scale-up is to finalize a
detailed large-scale process description with setting all
operational parameters and their ranges at scale
• Linear scale-up parameters include:
1. Temperature
2. pH
3. Pressure
4. DO level
5. Airflow rate
Temperature:
 Should be maintained the same at all scales
 Wide range for microbial processes but mostly 20 to 45
°C
 Large-scale operation should consider mixing time, heat
transfer and control precision
 Provide the same heat transfer rate in large-scale
reactors as in small reactors throughout the process to
maintain the temperature
 High density cultures e.g E. coli and yeasts require
extremely high heat removal rates
 Challenges to meet temperature control requirements
may be due to limited cooling capacity of large reactor
pH:
 Provides an important cell physiological condition
 Should be maintained the same at large-scale
 Tune automated pH control to provide the correct pH

regime
 pH may be influenced by acid/base concentration and

delivery system (pump, pressure, valves, etc)
Pressure:
 Should remain the same across all scales
 Pressure set points may be adjusted from small- to large-scale:
− by considering the higher hydrostatic pressure at large-

scale
− when small-scale process is developed at a location with a

different elevation thus the atmospheric pressure is
significantly different from that in large-scale region
− as a strategy or tool to help improve gas transfer at large

scales where a high agitation rate is difficult to reach
Dissolved oxygen (DO) level:
 DO set points are kept consistent at all scales to keep the

culture in the same respiration condition
 Defined automated DO control mechanism at large-scale
 DO is controlled in two ways:
a) In a cascade fashion in which agitation is increased based

on culture oxygen demand and then air flow rate is
increased or pure oxygen is introduced when agitation
reaches its maximum
b) Pressure may be increased to control DO

Airflow rate:
 Usually scaled up by a constant volumetric flow rate per
fermentation broth volume
 e.g. a 1-L fermentation with an airflow rate of 0.2 L/min
has an aeration rate of 0.2 vessel volumes per min (vvm)
 At a 100-L scale, the flow rate would be 20 L /min to
maintain 0.2 vvm
 A mass flow device is used to maintain accurate airflow
rate measurements at any scale
 In some cases, superficial gas velocity is kept constant
If Scale-Up Studies Fail!
• Should at this stage the fermentation process is
technically not feasible and there is a failure, there are
two options:
1. Either find the cause of the failure or go back to the

drawing board
2. Abort the whole project with minimum economic loss

to the investors
--END--

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School of Health Sciences &

CUBTM 219: Fermentation Biotechnology

Chapter 3: Fermentation Production Process Scale-up

Dr. R. Kamusoko (DPhil, MSc, BSc)

• Demonstrate understanding on the processes and

• Design a process scale-up for fermentation

• Scope of fermentation process scale-up

• Rules followed during scale-up

• Plan for process scale-up

• Cell banking strategy for large scale manufacturing

• Seed train scale-up strategy

• Production fermentation scale-up

• Linear scale-up parameters

• Microbial fermentation plays a critical role in many industrial

• Fermentation processes are utilised to produce a final product,

• Process scale-up enables a fermentation achieved in research

• Successful scale-up involves many aspects of preparation and

• To ensure that fermentation at large-scale is techno-

• Allows investigation of a product and process on small- and

• To get the same fermentation efficiency as obtained in small-

• Failing of process at industrial-scale causes million dollars loss

• Scaling-up gives investors more confidence on chances of

• Studies carried out at lab- or pilot-scale to yield data that

• The aim is to build industrial size fermenters capable or close

• A process developed in a lab (e.g. 0.5 – 10 L fermenters)

• Scale-up of large-scale processes is done in 2 stages:

1. Pilot-plant (100-10 000 L fermenters)

− to translate lab-scale process into a realistic scale-down

2. Demonstration plant (demo-scale) (10 000 to 100

− to minimise the risk of a large capital investment in the

1. Similarity in the geometry and configuration of

2. A minimum of three or four stages of increment in the

3. Each jump in scale should be by a magnitude or power

− Slight increase in the working volume will not yield

 Define product economics based on projected market size

 Conduct laboratory studies and scale-up planning at the

 Conduct preliminary larger than lab studies with equipment to

 Design and construct a pilot plant including provisions for

 Evaluate pilot plant results (product and process) including

1. Evaluating the results of lab studies, and making product

2. Producing small quantities of product for sensory,

3. Determining possible profitable by-products or waste

4. Providing data that can be used in making a decision on

5. Reduce the risk associated with construction of large

 Computer simulations and semi-empirical methods are

 Mathematical models are then tested in physical pilot

 Chemical similitude studies

 Finite elemental analysis

 Computational fluid dynamics

 Chemical similitude studies:

− Computer aided modeling and simulations in the

− e.g. Advanced Simulation Library

− The process of using various mathematical structures -

− The use of applied mathematics, physics and computational

− e.g. Sim scale, SciLab

 Finite elemental analysis:

− A computerized method for predicting how a product reacts to real-

• Mathematical models yield information about:

− Finalized mass and energy balance