4 Process Involved in Scale up

Criteria | Microbial Bioprocessing

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(1) Microbial Cell Process:

Theoretically the following criteria were assumed suitable basis
for scale up of bioreactors.
A. Constant power input per unit volume (P/V = constant).
B. Constant KLa
C. Constant mixing quality
D. Constant momentum factor (MF = ND. NWL (D W)) =
constant)
E. Similar drop size distribution (ds = constant)
F. Constant impeller tip speed ( NDi = constant)
G. Constant mixing rate number = (N/K)(Di/Dt) constant
Short accounts of scaling up of bioreactors of fermenters on the
basis of these criteria are discussed below;
1. Constant power input per unit volume:
For scaling up based on these criteria it is necessary to consider
whether it is gassed or non-gassed system. From the work of
Rushton and his associates, for geometrically similar, fully
baffled vessels with turbulent conditions, it may be noted that if
scale up should be based on maintaining a constant power

input per unit volume considering no gassing in the system one

may have the following.
(i) Non-gassed System:
When there is no gassing in the system then based on constant
P/V has the power number, Np as

It was also apparent that in an oil drop dispersion system

ds could serve as important criteria for scale up. The value of a
in equation 8.34 depends on the type of agitator used in the
system. For turbine impeller a= -0.6 whereas for draft tube
fermented its value ranges between -0.15 to -0.35.

6. Constant impeller tip speed:

It was rather surprising to recognize that most of the data on
impeller tip speeds collected were in the range 5-7 m sec 1
indicating the predominant importance of this parameter in
scale up. Specially in antibiotic production plants constancy in
tip speed ( NDi) was encountered in several cases. For scale up

In equation (8.37) N represents impeller tip speed and

Di1 impeller diameter
7. Constant mixing rate number:
In scale up of bioreactors mixing time is often used as criteria.
Mixing time (tmix), defined as the period of time required for the
homogeneous distribution of a small volume of pulsating
material in the bulk of the liquid, was used for scale up to adjust
proper mixing conditions in large vessels. However, limitation
of using tmix as scale up criteria is that it increases markedly with
the size of the vessel and it is difficult to maintain to constant in
both vessels at reasonable power expenditure.
In mixing a fermentation broth two vital questions to
be answered are:
(1) What size of motor is needed for a given impeller rotation
speed? and
(2) how long the broth must be mixed to achieve a required
degree of homogeneity?
From the present knowledge the first question can be answered
using Np NRerelations of Rushton and Bates et al. The answer

of the second question has been provided8 by introducing a new

dimensionless term mixing rate number [(N/K) (D/Dt)a], NMR in
the NP NRe profile to characterize the rate of approach to
uniformity in mixing vessels. This profiles as shown in Fig. 8.1 a
and b indicate that for large impeller Reynolds number (N Re >
104) the following correlations will be applicable depending on
the type of mixer used.
For turbine impeller mixer

(2) Scale Up of Filamentous Cell Fermentation:

Aerobic fermentative bioprocessing using filamentous
microorganisms cover a wide range of industrially important
bio-product formation systems. Various Streptomyces sp.
antibiotic fermentations fall in this group. Such bioprocessing
exhibit viscous and non Newtonian behavior following Power
law fluid characters.
These processes are typical and present considerable problems
in mixing and mass transfer for scale up purpose. Thus, agitator
power number (NP) and impeller diameter (Di) are important
case factors towards implications of Power law fluid on scale up
along with liquid volume (VL) and (HL /Dt) ratio.

Case 1:
Agitator power (Pt) number influence
a. Provided with same geometry, vs, (Pt/Vt), VL and
Di manipulation of volumetric mass transfer equations show

(3) Mammalian Cell Processing Scale Up:

In scaling up bioreactors of mammalian cell bioprocess most of
the correlations are based on laboratory models alone and still
awaits confirmation of their utility in industrial production
concern. For mammalian cell bioprocessing current industrial
scales typically involve bulk liquid volume of 25/ per day per
bioreactor for continuous or semi-continuous systems.
In batch processing this bioreactor volume ranges between
500-5000 I capacities, operating over 1-3 weeks. There are no
established scales up criteria for scaling up mammalian cell
culture process. Cell culture engineers therefore, still largely
depend on concepts and scale up procedures developed for
chemical or microbial/biochemical processes to begin with.
However, an engineering body of experience and data will
increasingly enable more specific design engineering and scale
up procedures for mammalian cell culture bioprocesses in
which kinetics is likely to be influenced by many factors and
associated with many problems and limitations.
In scaling up procedures specific to animal cell culture, effect of
scale on oxygen mass transfer through micro-porous
membranes like silicone tubes for supplying bubble free
oxygenation has been investigated. In order to aerate cell
cultures, the membrane is immersed in the medium and the gas
mixture (e.g. air, CO2 and O2) is passed through the tube under
pressure.
Depending on the difference in pressure between the gas and
the medium, gas flows through the membrane tube. Bubble free
aeration is achieved if the internal gas pressure does not exceed
the pressure at which bubbles will form. Correlation for bubble
free oxygenation through membrane tubes has been developed.
For this, novel bioreactors have been designed for cell culture
engineering in large quantities of the fragile complex
mammalian cells that synthesize commercially and medically

important proteins such as interferon and monoclonal

antibodies. The correlation for bubble free oxygen using
silicone micro-porous tube for oxygen mass transfer in this
novel bioreactor has been given by the following correlations.

In these correlations, Sh is Sherwood number, K is film mass

transfer coefficient (m s_1), d is tube diameters, D0 is molecular
diffusivity of oxygen (m2 s-1), e, f, g are specific constants, Dt is
bioreactor diameter (m) and NRe is impeller Reynolds number.
The provision of an adequate oxygen supply to large volumes of
mammalian cells is the most crucial barrier to scale up,
especially in suspension systems. Oxygen is only sparingly
solube in cell culture medium (0.2 m mole O2I-1). Oxygen
demand of a culture (106 cells per ml) ranges between 0.053 m
mole O2 l-1 h-1 and 0.59 m mole O2I-1 h-1 depending on the type of
cell. Scale up of mammalian cell cultures are, therefore, bound
by several barriers as shown in Table 8.1.

As d, D0, NRe and Dt are given and e, f and g are determined

experimentally, then equation 8.48 enables calculation of the
oxygen mass transfer coefficient from impeller speed. With the
assumption that gas velocity and composition within the
silicone tube are constant, one may calculate the power

required for this form of aeration at the large scale using the
correlation given below.

In this correlation P, is the power input per unit mass of liquid

(W.kg-1), V is the reactor volume (m3), D; is impeller diameter
(m), p is the kinematic liquid viscosity (m2 s-1) and NP is the
power number. Thus, scale up might be carried out on the basis
of impeller speed to yield a constant oxygen mass transfer rate,
but the power input per unit mass will vary with scale. It will
need to check the power against an upper limit which will be
determined by the shear sensitivity of the mammalian cells.
Lavery and Ninow in their study of the conventional power
input versus mass transfer relation in sparged mammalian cell
culture indicated the necessity of consideration of both the
effect of head space aeration and the effect of added antifoam.
This could account for the deviations from the expected values
of KLa in their result.
A number of serious problems associated with the design and
scale up of high density perfusion cultures of mammalian cell
bioprocessing have been pointed out. For overcoming the
associated problems in cultivation of animal cells and their
scale up the following usage has been suggested.
1. Micro bubble technology and soluble surfactants for meeting
cellular oxygen uptake rate (OUR).
2. Turbine or low shear impellers for achieving well mix
cultures, such that it provides independent requirements of
mixing and bubble aeration.
3. Minimization of ammonia accumulation by low glutamine
supplemented media design.

4. Appropriately sized microfiltration hollow fibre cartridges

which are steam sterilizable and easily cleaned after repeated
uses.
(4) Plant Cell Processing Scale Up:
A typical plant cell culture manufacturing process is shown in
Fig. 8.2. The great upsurge of interest in process biotechnology
in the 1970s carried plant cell culture with it. This is primarily
because of centuries plants have been an important source of
drugs and chemicals.

The extensive effort in development of plant cell culture is

evident from the literatures on the subject. As plant cell
cultures have many differences in comparison to microbes the
design and scale up of suspension cultures bioreactors for plant
cells also differ to a great extent. This becomes more
pronounced as plant cell cultures can achieve high cell densities

and viscosities. In such systems need for good mixing must

have a compromise to shear sensitivity of plant cells.
Thus, in plant cell culture scale up system information is very
important. Useful system informations for scale up of plant cell
culture as proposed by Goldstein are presented in Table 8.2. In
addition to these quantization of entities as effected through
bioprocess engineering studies at laboratory scale needs
information on operational variables as listed in Table 8.3.

The first consideration in generating these informations is the

selection of a suitable bioreactor type. Continuous stirred tank
bioreactor (CSTBR), air lift bioreactor (ALB), immobilized plant
cell bioreactor etc. have been used in plant cell cultivations
either in batch, plug flow or in continuous modes. In these
three modes culture residence time is analyzed based on the
relations given in Table 8.4. Assuming autocatalytic growth
these relations emerge from classical catalysis.

The process parameters for scale up of plant cell cultures as

listed in Table 8.5 demand low shear bioreactor for plant cell
aggregates. As reported in literature plant cell cultures often
form aggregates in suspension cultures in bioreactor. It can
make the provision of oxygen difficult: the high aeration or
mechanical agitation necessary can both cause shear damage to
the cells.
Table 8.5 Process parameters for scale up of plant cell
culture:

For overcoming the above difficulty and to facilitate scale up of

plant cell culture bioreactor a design of low shear bioreactor has
been reported. This design is stated to create separate cell
growth and aeration zones. The bioreactor design is analogous
to fluidized bed columns except that the cells are immobilized
naturally (by aggregation).
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Home / Articles / 2005 / Don't Get Mixed Up By Scale-Up

Don't Get Mixed Up By Scale-Up

Use this systematic approach to successfully
scale-up mixing processes even without
geometric similarity.
By David S. Dickey, MixTech, Inc.
Aug 04, 2005
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- A A A A +
A production mixing unit is usually not geometrically similar to the mixer used for process
development. Such differences can make scale-up from the laboratory or pilot plant
challenging. A solution to these problems is to systematically calculate and evaluate mixing
characteristics for each geometry change.
Geometric similarity is often used in mixing scale-up because it greatly simplifies design
calculations. Geometric similarity means that a single ratio between small scale and large
scale applies to every length dimension (see figure). With geometric similarity, all of the
length dimensions in the large-scale equipment are set by the corresponding dimensions in
the small-scale equipment. The only remaining variable for scale-up to large-scale mixing is
the rotational speed one or more mixing characteristics, such as tip speed, can be
duplicated by the appropriate selection of a large-scale mixer speed.

The two most popular and effective geometric scale-up methods are equal tip speed and
equal power per volume. Equal tip speed results when the small-scale mixer speed is
multiplied by the inverse geometric ratio of the impeller diameters to get the large-scale
mixer speed:
N2 = N1(D1/D2)
Equal power per volume involves a similar calculation, except the geometry ratio is raised to
the two-thirds power:
N2 = N1(D1/D2)(2/3)
This expression for power per volume only applies strictly for turbulent conditions, where the
power number is constant, but is approximately correct for transition-flow mixing.
Another important mixing characteristic is torque per volume, which often represents mixing
intensity, in terms of fluid velocities. While torque is just power divided by speed, torque per
volume is similar to momentum transfer and is closely related to the effective motion created
by the mixer. For geometric similarity and turbulent conditions, torque per volume reduces to
the same scale-up formula as equal tip speed (Eq. 1). Most of the practical scale-up rules for
geometric similarity, including equal solids suspension, fall in the speed range between
equal tip speed and equal power per volume. Other criteria, such as equal blend time or
surface motion are difficult to scale-up because of the rapidly increasing mixing intensity for
large-scale tanks.
The sidebar gives formulas for calculating mixing variables in conventional U.S. engineering

units. Derivations and details on the various equations for geometric scale-up and mixer
evaluation appear in many references such as widely available handbooks [Refs. 1 and 2].
Five steps
The recommended approach for doing non-geometric scale-up is as follows:
1. Calculate the laboratory- or pilot-plant-scale values for the mixing variables listed in the
sidebar.
2. Do a geometric-similarity scale-up to the large-scale tank diameter. Use a criterion such
as tip speed to set the large-scale speed. Calculate the mixing variables for the new, largescale conditions.
3. As needed, change the impeller diameter at the large scale. Calculate the mixing
variables and, if required, modify speed to keep an important variable such as tip speed or
torque per volume constant.
4. Switch impeller type, if desired or necessary, and recalculate mixing variables using the
appropriate power number for the new impeller. Calculate the mixing variables and make
decisions or adjustments as required.
5. Alter the liquid level, if necessary, to match volume specifications at the large scale.
Calculate the mixing variables again for the new liquid level. If undesirable changes occur,
adjust mixer speed or number of impellers.
The key to success is making simple step-by-step changes in the design and calculating
important variables each time. In this way, you can evaluate the cause and effects of
changes and make corrections when necessary.
Applying the approach
Suppose for example, we did a series of experiments with a 5.0 gal. batch in an 11.5 in.-dia.
pilot reactor, which had two 5-in.-diameter, three-blade, pitched-blade turbines, and found
that we could get good results with the mixer operating at 300 rpm. The material we were
mixing had a final viscosity of 85 cP and a specific gravity of 1.05. Rearranging the volume
formula in the sidebar and plugging in values, we can find the liquid level for 5.0 gal. in a
11.5 in.-diameter cylindrical tank:
H = 4 (231)(5)/p(11.5)2 = 11.12 in.
The 11.12-in. liquid level assumes that the tank bottom is flat and the tank internals
(impellers, shaft, baffles, etc.) do not take up any space. In the real world, the tank may have
a dished bottom and the internals typically will occupy 2% to 5% of the volume for this type
of reactor. While corrections can be made for bottom shape and tank internals, for simplicity
we will ignore these corrections here.
With this information, we can calculate the operating conditions that exist in our pilot-scale
experiment (see table). At these conditions, the Reynolds number of 995 would suggest that
the mixer is operating in the transition regime, which applies for 1 < NRe < 20,000. For a
pitched-blade turbine the power number, which remains constant in the turbulent range, will
begin to increase in the transition and viscous regime. For simplicity, lets assume that the
power number does not begin to increase until the Reynolds number drops below 900. Had
the Reynolds number been less than 900, we would have to make an appropriate correction
to the power number and use that corrected number to calculate power and torque
characteristics of the mixer. This additional step to correct power number should be done for
each subsequent step in the scale-up process if the Reynolds number suggests that such a
correction is necessary. (Power number corrections differ with type of impeller.)

Now suppose we want to duplicate our process results in an 84.0 in.-dia. tank with a 3,000
gal capacity using four-blade, pitched-blade turbines. As we will see while doing the
calculations, the tank geometry and impeller type both change in the scale-up process. Such
changes are common. In this particular case, a combination of experience, literature
research and experimentation leads us to believe that the tip speed should be held constant
when we scale-up and that torque-per-volume may represent mixing intensity.
As a first step in our scale-up calculation, we will use geometrical similarity to do a scale-up
from our 11.5 in.-dia. pilot tank to an 84-in.-dia. process tank. Geometric similarity means
that our scale-up length ratio of 84/11.5 = 7.3 will multiply all of the length dimensions. Thus,
the 5.0 in.-dia. pilot turbine will scale-up to a 36.5 in.-dia turbine in the large-scale tank and
the liquid level will be 7.3 11.12 = 81.2 inches. However, this liquid level provides a volume
of only 1,948 gal. We will need to adjust the liquid level later to get our desired results.
To maintain the same tip speed in the large-scale mixer as in the pilot unit, we must adjust
rotational speed because the impeller diameters differ and impeller diameter times rotational
speed gives tip speed. So, for equal tip speed, the operating speed for the large-scale mixer
is:
300 (11.5/84) = 41.1 rpm
At 41.1 rpm, we can do the same calculations we did for the pilot-scale results.
As shown in the table, geometric scale-up with constant tip speed increased the Reynolds
number, power and torque. However, power per volume decreased and torque per volume
remained constant. For this first step in our scale-up process, both tip speed and torque per
volume remained constant, which satisfies our process scale-up objectives.
The next step
At this point we will change to the four-blade turbines required in our large-scale process.
The four-blade turbines have a power number of 1.37 instead of 1.19; so we can round down
the turbine diameter to 36.0 in. and round up the speed to 42.0 rpm. Such adjustments seem
both reasonable and practical. However, we should recalculate our mixing results to be sure.
With this turbine change, tip speed essentially stays constant and our torque per volume
rises by a little more than 20%. This increased torque per volume represents a possible
boost in apparent mixing intensity, which could be judged to be beneficial, because the
larger tank will blend more slowly than the small pilot tank.
Adjust the tank volume
Now we need to increase tank volume to the required 3,000 gal. for the full-scale process.
We can achieve this by raising the liquid level to 125 in. from the geometric value of 81.2 in.
We must check the effects of the larger volume on the mixing variables.
Because the volume changed but not the mixer, only the power per volume and torque per
volume changed. The torque per volume is now only 80% of our original pilot and geometricscale-up values, which might translate into too little mixing and thus uneven results.
Several possible remedies exist for the undesired reduction in torque per volume. Suppose
that with the volume increase we had held both the tip speed and torque per volume
constant. For such a scenario, both the turbine diameter, D, and rotational speed, N, must
change as the volume increases. To keep tip speed constant, we can use the relationship:

NDvTIP
Then, per Eq. 1, for constant tip speed:
N2D2 = N1D1
For torque per volume, the following relationship based on turbulent conditions applies:
t/V N2D5/V
For constant torque per volume:
N22D25/V2 = N12D15/V1
Factoring tip speed out of the constant torque per volume relationship gives:
(N22 D22) D23/V2 = (N12 D12) D13/V1
which for constant tip speed can be reduced to:
D23/V2 = D13/V1
Rearranging Eq. 7 and solving for the new turbine diameter gives:
D2 = D1(V2/V1)1/3 = 36(3000/1948)1/3 = 41.6 in.
Using the new turbine diameter, a speed can be selected to satisfy the constant tip-speed
condition:
N2 = N1(D1/D2) = 42 (36.0/41.6) = 36.3 rpm
According to the algebra, this larger turbine and lower speed should keep both tip speed and
torque per volume constant; this can be checked by recalculating mixing values.
By similar methods other combinations of variables, such as tip speed and power per
volume, could be held constant. Also, impeller type can be changed and mixing variables
held constant by including the power number in the expressions for power or torque. Many
different adjustments to mixer characteristics can be handled by similar expressions and
manipulations. Usually only a couple of mixing variables can be held constant while the
others change. Sometimes this will lead to impractical solutions such as extreme impeller
diameters or rotational speeds. If viscosity, through Reynolds number, affects power number,
the exact mathematical relationships do not hold for power and torque. In those cases
estimate approximate values and adjust calculated mixing values if necessary.
Returning for a moment to the volume change that was necessary in going from the
geometrically similar scale-up to the actual large-scale volume, another possibility should be
considered. The geometric scale-up resulted in a volume of 1,948 gal., yet the large-scale
process required 3,000 gal. The ratio of the volumes is almost exactly 2:3, so why not
consider going from two turbines to three turbines. The resulting changes in the calculated

values are shown in the table. This solution to the scale-up problem not only meets the
large-scale process requirements but also maintains the desired characteristics of the pilotplant situation.
Avoid mix-ups
As we have seen, taking successive steps allows the development of alternative solutions to
scale-up. Similar methods can be used to scale-down process problems for investigation in a
pilot-plant or laboratory simulation. Here, too, non-geometric similarity often is a problem.
Such scale-down calculations should help pinpoint appropriate operating speeds to test in
the small-scale mixer.
In any scale-up or scale-down evaluation, some variables can be held constant while others
must change. For example, even with geometric similarity, scale-up will result in less surface
per volume because surface area increases as the length squared and volume increases as
length cubed. Similarly, keeping blend time constant rarely is practical with any significant
scale change. Larger tanks take longer to blend than smaller ones. Also, Reynolds number
is expected to increase as size increases. In addition, standard operating speeds or
available impeller sizes may necessitate a final adjustment to the scale-up calculations.
Rules for scale-up always have exceptions but understanding the effects of scale-up,
especially non-geometric scale-up, can provide valuable guidance. Indeed, appreciation of
the tradeoffs involved in non-geometric scale-up may be crucial for success with large-scale
mixing processes.
Dr. David S. Dickey is senior consultant at MixTech, Inc., Dayton, Ohio. He teaches a short
course on pilot-plant scale-up methods for industrial mixing for the University of Wisconsin.
E-mail him atd.dickey@mixtech.com.
REFERENCES
1. Perrys Chemical Engineers Handbook, 7th ed., Green, D.W. and J.O. Maloney, Eds.,
McGraw-Hill, New York (1997).
2. Handbook of Industrial Mixing, Science & Practice, Paul, E.L., et al., Eds., Wiley,
Hoboken, N.J. (2004).
NOMENCLATURE
D = impeller diameter, in.
H = liquid level, in.
N = rotational speed, rpm
P = power, hp
T = tank diameter, in.
V = volume, gal
v = velocity, ft/min
= viscosity, cP
= specific gravity
KEY EQUATIONS
Impeller power, hp
Power per volume, hp/1,000 gal
Power number

P = NPN3D5/(1.524 1013)
PV = 1,000 P/V
NP = (1.524 1013)P/r N3 D5

Reynolds number
Tank volume, cylindrical tank, gal

NRe = 10.74 D2N/m

V = (/4)(T2H/231)

Tip speed, ft/min

vTIP = DN/12

Torque, in.-lb

t = 63,025 P/N

Torque/volume, in.-lb/1,000 gal

tV = 1,000 t/V

Help for calculating the impeller tip speed to avoid

excessive erosion due to suspended particles
Applets are programs based on the java language that are designed to run on your computer using the
Java Run Time environment.

The maximum impeller tip speed is based on tests that were done by the Hydraulic Institute www.
pumps.org. You can find more information in The Hydraulic Institute's Centrifugal Pumps for Design
and Application ANSI/HI 1.3 standard.

This applet is designed to help you calculate the tip velocity of an impeller vane with the simple v = wr
formula.

The recommendations on the maximum tip speed are as follows:

- dirty water - 130 ft/s
- medium slurries up to 25% solids and 200 micron solids - 115 ft/s

- higher slurry concentrations and larger solids - 100 ft/s

- pumps with elastomer impeller - 85 ft/s

Tip speeds that are higher than recommended are likely to cause excessive erosion. Elastomer
impellers are often used in the mining industry for the transfer of tailings which is a highly abrasive
slurry.

The equation used for calculating tip speed is:

where
v: velocity
D: impeller diameter
RPM: revolutions per minute

Scaling up may involve optimization of culture at shake flask level using suitable carbon
source such as glucose, fructose, glycerol etc. along with the medium, micro and
macronutrients and a buffering system to maintain pH at high cell density cultures.
Shake flask culture optimization is pre-requisite for starting any culture studies at fermentor
level. Optical densities attained and the expression levels of the protein of your interest
should be the criteria for the selection of suitable carbon source.
Media can be of 5x, 10x, 25x or higher concentrations, depending upon your ability to make
and handle the medium at fed-batch runs.

At fermentor level you can set up a batch-culture run, or a fed-batch culture run. Before
running a fed-batch culture it is advisable to run a number of batch runs to determine the
growth pattern and maximum optical densities (ODs) of your culture. Once the growth
pattern is known running a fed-batch culture is easier as the method and time of feeding are
now the main concern to attain the success in a fed-batch run.

Feeding strategies will depend upon the controlling efficiencies of both the personnel and the
fermentor. Commonly used ones are exponential feeding, pH-stat, DO-stat. In exponential
feeding nutrient medium is fed with the exponential growth requirements of the culture,
where as in pH-stat and DO-stat, nutrient medium is fed but dissolved oxygen requirement or
pH is maintained without affecting much variation ie., constant or static pH or DO state is
maintained. Exponential feeding is slightly difficult to handle during log phase and high ODs
where as pH-stat and DO-stat are easier and no calculations are required for determine
feeding rate, it can be controlled manually.
OD is to be recorded periodically preferably every 1 hr period. To know the growth pattern of
the culture.