B I O P R O C E S S TECHNICAL

Lessons in Bioreactor Scale-Up, Part 2

A Refresher on Fluid Flow and Mixing

LY
Muhammad Arshad Chaudhry

N
C

O
ommercial-scale production of components. Such force must be applied Figure 1: Familiar examples of laminar
recombinant therapeutic proteins to produce fluid flow (1). Scientists and turbulent flow
routinely involves suspension categorize fluids according to their

N
Turbulent flow
cultures of mammalian cells in density and viscosity as
bioreactors with up to 10,000 L capacity. • compressible (or incompressible) —

IO
With advances made over the past 30 whether their density changes (or not)
years in cellular engineering, basal and in response to pressure, with gases
feed media development, and bioprocess generally being compressible and

S
engineering, expression titers of ~10 g/L, liquids being incompressible
viable-cell densities of >3 × 107 cells/mL, • viscous (all fluids have a finite

IS
and specific productivities of >20 pg/cell/ viscosity, which is responsible for Laminar flow
day are now common. Such high cell “internal friction” during flow,
M
densities (with the potential to go even otherwise expressed as a fluid’s
higher) increase mixing and aeration resistance to motion)
demands and can subject cells to • Newtonian or non-Newtonian, those layers, and that influences the
R
aggressive environments such that they depending on whether they obey velocity of their flow. Particles moving
experience high hydrodynamic stresses. Newton’s law of viscosity. from a slow-moving layer to a faster
E

This installment of my bioreactor Shear stress is calculated by dividing layer will act to reduce its speed; those
scale-up series should refresh your the shear force applied to a fluid by the coming from a fast layer to a slower one
P

knowledge of fluid flow, mixing, and cross-sectional area to which it is will have an accelerating effect. Thus
mass transfer in bioreactors, including applied. Strictly speaking, shear stress are shear stresses induced by fluid flow
H

how the interplay of those parameters is one of two types of fundamental as a consequence of velocity differences.
creates the environment that cells hydrodynamic forces, the other being Depending on fluid velocity, flow can
IT

experience. That in turn influences their “normal stress.” Both are determined be characterized as laminar or turbulent
growth, metabolism, and protein commonly as viscosity multiplied by the (Figure 1). When the overall fluid motion
production. velocity gradient, with flow direction is slow, adjacent fluid layers move in an
W

being perpendicular for shear stress and orderly (more or less parallel) motion
Fluid Flow in Bioreactors parallel for normal stress. Hence, a called laminar flow. In a fast-moving
Fluids — either liquids or gases — are velocity gradient for shear stress is fluid, particles and parcels of fluid
T

substances that undergo continuous written as dUx/dy and for normal stress frequently cross layers randomly and
deformation when subjected to shear as dUx/dx, where U is the velocity of chaotically in a turbulent flow.
IN

force. Shearing causes deformation, a fluid and x and y represent the direction Reynolds number (Re) is a
change in the relative positions of fluid of the force/stress. dimensionless variable for characterizing
R

When fluid flows through a pipe or fluid motion as laminar or turbulent.

over a solid object, the velocity of that Equation 1 calculates Re for fluid flow in
PRODUCT FOCUS: All biologics
P

fluid changes depending on position. pipes with circular cross-section:

PROCESS FOCUS: Production The fluid layer in contact with the wall Re = DUρ/μ
E

of a pipe, for example, is assumed to where D is the pipe diameter, U is

AUDIENCE: Process development have the same velocity as the wall or fluid velocity, and ρ and μ are density
R

and manufacturing solid object (zero velocity). The next and viscosity of the fluid, respectively.
fluid layer inward has a bit higher Because the biopharmaceutical
KEYWORDS: Impellers, Reynolds
velocity because of the overall fluid industry is concerned primarily with
number, power number, shear,
motion, the next layer inward has more, fluid flow in stirred-tank bioreactors,
distribution, dispersion, and diffusion
and so on. Within every fluid is a the relevant Reynolds number is defined
LEVEL: Basic continuous exchange of particles among by Equation 2:

32 BioProcess International 22(6) JUNE 2024

Table 1: Comparison of parameters used for calculating hydrodynamic stress in two Figure 2: Dimensions of mixing-related
different 1000-L single-use bioreactors (adapted from Reference 2) components in a bioreactor.
1,000-L Single-Use Bioreactors Impeller
Parameter Units XDR1 HyPerforma S.U.B.2 motor
Revolutions per minute (rpm)* min –1 108 95
Liquid volume (VL) m3 0.783 0.762
Volume of air sparged per unit volume T = tank diameter
L/L/min 0.029 0.043
of growth medium per minute (vvm)*
Energy dissipation rate (EDR) mean* W/m3 28 30
Head
EDR maximum W/m3 2.3 × 103 6.2 × 102 space
Intensity (λ κ)** μm 78 76
1 from Cytiva 2 from Thermo Fisher Scientific Baffles

H = tank height
* Maximum values during a process at the current bioreactor size ** Calculated from EDR mean

HL = fluid height
Re i = Ni Di2ρ/μ direction, but superimposed on that are Di = impeller diameter
where Ni is the stirrer speed, Di is the secondary chaotic movements of fluid
impeller diameter, and ρ and μ are particles causing flow deviations and

C = impeller
clearance
density and viscosity of the fluid, generating vortices of varying shape,
Sparger
respectively. size, speed, and rotational direction. disc
Conceptually, the Reynolds number Those irregular secondary motions in
expresses the ratio of inertial to viscous turbulent flow have profound
forces. During laminar flow (low consequences for mixing efficiency,
Reynolds number), viscous forces energy loss, and shear intensity. (NRei > 104), the size of the smallest
dominate, and fluid motion is slow. Turbulent flows develop spinning or eddies is given approximately as the
However, with increasing Reynolds swirling fluid structures called eddies, Kolmogorov scale of mixing (λ) or scale
numbers, inertial forces become more which can stretch, coalesce, and divide. of turbulence, defined by Equation 3:
dominant, and flow transitions from Fluid velocity within an eddy undergoes λ = (ν3/ε)1/4
laminar to turbulent — then becomes great change in magnitude and where λ is the characteristic length
fully turbulent at sufficiently high Re direction within a relatively short of the smallest eddies, ν is the
values. distance and time. kinematic viscosity of the fluid, and ε is
Pipe flow is laminar if Re < 2100. For Eddies of different sizes appear in the local rate of turbulent energy
Re values of 2000–4000, the flow is turbulent flow. The size of the largest dissipation per unit mass of the fluid.
considered to be in a transition state eddies is limited by the boundary of the At a steady state, the average rate of
from laminar to turbulent. It is fully flow system. Hence, the diameter of the turbulent energy dissipation throughout
turbulent once Re > 4000. In stirred largest eddies in a bioreactor is similar a bioreactor is equal to the power input
tanks, the value of Re that marks the to (but somewhat smaller than) the by the impeller to the fluid within the
transition from laminar to turbulent flow diameter of the reactor vessel. Large tank. Hence, the greater the power input
depends on tank and impeller geometry. eddies are unstable and give rise to to the impeller, the smaller the eddies
smaller ones, which in turn produce will be. Because λ depends on viscosity,
Turbulent Flow in Bioreactors even smaller eddies, and so on. In a smaller eddies are produced in low-
Turbulent flow is the flow regime that stirred bioreactor, large eddies derive viscosity media for a given power input.
cells encounter in bioreactors. Turbulence energy from the bulk flow generated by An energy input of 0.1 W/kg, which
is essential for effective mixing (mass the impeller, so they contain most of the currently is considered to be high for
and heat transfer in fluids), so achieving turbulent kinetic energy. The smallest animal-cell cultures, yields an estimate
turbulent-flow conditions in bioreactors is eddies cannot sustain rotational motion for the smallest eddy diameter λ of about
vital to culture success. As noted above, and lose their energy as heat through 0.06 mm (60 μm). That represents the
the Reynolds number indicates when the effects of viscosity and fluid friction. smallest scale of mixing achievable by
turbulent flow is generated. As that Maintaining turbulent flow thus requires dispersion under such conditions.
number increases, inertial forces come to continuous supply of energy to make up The characteristic length of the
dominate viscous forces in the fluid, for what is lost as heat (1). smallest eddies (λ) often is an indicator
overcoming the tendency of viscous Dispersion is the process of breaking of the potential for cell damage caused
effects to dampen flow instability. Thus, up the bulk flow into smaller and smaller by fluid shear. Experiments have shown
turbulence can be regarded as a highly eddies to facilitate rapid transfer of that if biological entities (e.g.,
disordered fluid motion resulting from material throughout a tank. The degree mammalian cells) are smaller than λ in a
instabilities growing in an initially of homogeneity possible as a result of bioreactor, then shear damage to such
laminar flow field (1). dispersion is limited by the size of the entities will not occur. Minow et al.
Turbulent flow is very complex by smallest eddies that form. Assuming that presented a comparative study of mixing
nature, so our understanding of it is far isotropic turbulence is prevailing in a and mass transfer in two different
from complete. The flow has an overall stirred tank and that flow is turbulent 1000‑L single-use bioreactors. The

JUNE 2024 22(6) BioProcess International 33

Figure 3: Pitched-blade and paddle (Rushton) impellers commonly used in bioreactors
(adapted from Reference 3) equipment has a significant effect on
Rushton
agitation efficiency, directly affecting
Pitched-blade
impeller paddle power requirements and operating costs.
impeller Development of hydrodynamic forces in
media and other fluids drives aeration
processes such as gas-bubble break-up,
gas hold-up, and uniform dispersion of
bubbles. Thus, mixing can determine the
success a bioprocess.
Bioreactor Geometry: Most
stainless-steel (or reusable) large-scale
bioreactors are cylindrical, with a ratio of
tank height to diameter (H/T) >1 (Figure
Figure 4: The axial flow pattern induced by pitched blade impellers (left) and the 2). Baffles — cylindrical metal strips —
turbulent kinetic energy distribution profile of a pitched-blade impeller (right) generated
are mounted against a bioreactor wall to
by computational fluid dynamics (CFD) simulation (adapted from Reference 1).
reduce gross vortexing and swirling in
the liquid. Mixing is achieved by means
Baffle of an impeller mounted on a centrally
k (m2s–2) located stirrer shaft driven by a motor.
0.00050
The rotating impeller pumps and forces
liquid away from it, creating a regular
0.00015
flow pattern that circulates through the
vessel and periodically returns to the
impeller region. For aeration, gas
(usually air and/or oxygen) is introduced
into the vessel by means of a sparger
located beneath the impeller. A
authors calculated the Kolmogorov eliminating gradients of temperature, headspace typically falls in the range of
length of the smallest eddies prevailing concentration, and other properties (1). 20–30% of total vessel volume for
in each bioreactor type (2). During mixing, material interchanging cylindrical bioreactors.
As Table 1 shows, the calculated among different locations of a vessel The shape of the base of a stirred
values of λ for each bioreactor type were (such as a mixing container or a tank affects mixing efficiency. Most
close to 80 μm — far larger than the bioreactor) causes components to stirred bioreactors have a rounded rather
15-μm size of Chinese hamster ovary “mingle.” Mixing is widely used in than a flat base, preventing sharp
(CHO) cells used in the study. Similarly, bioprocessing for corners and pockets into which fluid
Godoy-Silva et al. described CHO cells • blending soluble components of currents could penetrate, and thus
that could withstand power inputs powder media in water to prepare for discouraging the formation of stagnant
≤6000 W/kg (3). At power dissipation cell culture zones. The energy required to keep
rates >300 W/kg, which is still much • dispersing gases such as air and solids (such as cells) suspended in a
higher than the power densities in oxygen through liquids in the form of stirred tank also depends on the shape
commercial cell-culture reactors, the only bubbles of the vessel base. For efficient mixing
impact observed was in the glycosylation • maintaining suspension of solid with a single impeller, the impeller
pattern of expressed proteins, but the particles such as cells or cellular diameter (Di) should fall between 0.25
cells were not affected (7). Even if such aggregates (e.g., on microcarriers) and 0.50 of the tank diameter (T), and
results are specific to the system studied, • promoting heat transfer to or from the liquid height (HL) should fall within
they still show that shear damaging has liquids. 1.0 and 1.25 of T. That’s because mixing
been overestimated in recent decades. Mixing is one of the most important intensity decreases rapidly as fluid
Nienow arrived at the same conclusion in unit operations in bioprocessing. It is not moves away from the impeller zone, so
two different papers (4, 5). Hence, it is enough to just fill a bioreactor with large liquid volumes in the upper parts
safe to say that eddies <10–20 μm nutrient-rich media and cells. Unless the of a vessel (away from the impeller) are
(smaller than the typical mammalian cell contents of that bioreactor are mixed, difficult to mix — and thus should be
size) possess very little turbulent kinetic cells will settle, and zones of nutrient prevented. Another aspect of vessel
energy, whereas larger eddies merely depletion will develop as they rapidly geometry that influences mixing
carry cells along convectively (8–12). consume the material they need within efficiency is the clearance (C) between
their local environment. Mixing controls the impeller and the tank bottom, which
Mixing in Bioreactors their access to dissolved nutrients and affects solids suspension, gas dispersion,
Mixing is a physical operation that oxygen while playing a critical role in and hydrodynamic stability. For most
reduces nonuniformities in fluid by controlling culture temperature. Mixing stirred bioreactor systems, the ratio of

34 BioProcess International 22(6) JUNE 2024

Figure 5: The radial flow pattern induced by Rushton impellers (left) and their
turbulent kinetic energy distribution profile (right) generated by computational fluid blade configuration they also can balance
dynamics (CFD) simulation (adapted from Reference 1) both axial and radial flow patterns (4, 5).
Axial flow is necessary for top-to-
Baffle bottom mixing in a stirred-tank
bioreactor, and pitched-blade impellers
k (m2s–2)
(with blades angled <90° to the plane of
1.0
0.8
rotation) can generate such flow
0.6 efficiently. As Figure 4 shows, fluid
0.4 leaving an impeller is driven
0.2 downwards until it is deflected from the
0.0
bottom of the vessel. Then the fluid
spreads out and flows up along the
vessel walls before it is drawn back to
the impeller. The rotational flow makes
Figure 6: Determination of mixing time (tm) by following the concentration (C) response baffles necessary to break down vortex
after a tracer dye is injected into a stirred tank (adapted from Reference 1); Cf = final formation along the vessel walls.
concentration, Ci = interim concentration, tc = circulation time A pitched-blade impeller agitating at
sufficiently high velocity in a low-
viscosity medium (e.g., water and cell
culture media) generates a turbulent
region of high shear and rapid mixing
tc tc tc near the impeller. That high-shear
Trace Concentration

region is responsible for bubble break-up

0.1 (Cf – Ci )
in sparged bioreactors. As Figure 4
shows, the distribution of turbulent
Cf kinetic energy is not uniform
throughout the tank, with the highest
values mostly occurring in a region
close to the impeller. Levels of turbulent
kinetic energy in the remainder of the
Ci tank away from the impeller are up to
tm an order of magnitude lower than the
Time maximum values measured (1).
Axial flow is useful when strong
vertical currents are required — e.g., to
clearance to fluid height (C/HL) typically Fluid velocity is highest at the tip of suspend solids or cells in fluid. The
falls into the range of 0.33–0.66 (3). a rotating blade (or paddle), and it ability to operate a pitched-blade
Impeller type is critical to bioreactor rapidly decreases away from the impeller in either up-flow or down-flow
design. An impeller system must be able impeller. As it rotates, the impeller mode is an important requirement that
to impart enough power into the cell induces rotational velocity in the can be provided by a bidirectional motor.
culture medium for enabling good surrounding fluid, which can be For most animal cell-culture systems,
mixing efficiency, homogeneity, and observed by the high velocity region these impellers are programmed for
sufficient mass transfer to satisfy next to the impeller. Rushton impeller up-flow pumping mode. However, the
cellular oxygen demand. Figure 3 shows paddles also induce radially outward down-flow option can be useful for
two of the most common impeller flow in nearby regions. It pulls liquid enabling suspension of adherent-cell
designs used in both reusable and from top and bottom regions of a cultures on microcarriers at low speeds
single-use bioreactors: Rushton and bioreactor, which creates a circular with low specific power input to prevent
pitched-blade impellers. When the blade motion inside the vessel. That motion shearing the cells from their carriers (3).
(or paddle) of an impeller rotates, it enhances fluid mixing. By contrast, Disc-mounted impellers with six flat
pushes the adjacent fluid. That action pitched-blade impellers create mainly blades, commonly known as Rushton
creates a high-pressure region on the an axial flow (from top to bottom). turbines, generate radial flow (Figure 5).
front face and a low-pressure region The preferred impeller design for cell- They are used frequently in microbial
immediately behind the blade (or culture bioreactors uses pitched blades fermentation, for which they can
paddle). Those regions are responsible (with optimal blade diameter and generate very high shear and thus
for fluid motion inside a bioreactor and agitation speed) because paddles are provide effective gas dispersion. A
eventually for fluid mixing. Fluid more likely to cause shear damage. Rushton turbine generates a jet of high-
located away from the impeller generally Pitched-blade impellers primarily speed flow radially outward from the
remains at a constant pressure. generate axial flow, but with appropriate impeller toward the walls of the tank.

36 BioProcess International 22(6) JUNE 2024

 Figure 7: Determination of mixing time (tm) in a 400-L bioreactor fitted with either
That fluid stream gets divided in half, three Rushton turbines (circles) or three pitch-blade impellers (triangles) (adapted from
with one stream flowing up and the Reference 14); data are fitted to the correlation reported by Ruszkowski (15); D =
other down toward the tank bottom. impeller diameter, H = tank height, T = tank diameter, ε = the local rate of turbulent
energy dissipation per unit mass of the fluid, θ 95% = time needed to reach 95%
Bulk flow in the vessel thus comprises
homogeneity in the bioreactor.
two large ring vortices, one above and
100
the other below the impeller.
Another characteristic of Rushton
Θ95% = 17.92/3 · ε̄ –1/3 · (D/T )–1/3
turbines is their generation of trailing
vortices in fluid behind the horizontal

Θ95% (seconds)
edges of each flat impeller blade (Figure
5). The vortices play a critical role to
determine mixing performance because
most mixing takes place near the
vortices issuing from the impeller blades. H/T = 1.60
These trailing vortices also provide gas D/T = 0.33
Θ95% = 10.52/3 · ԑ̄
ε –1/3 · (D/T )–1/3
dispersion in sparged systems. The
associated steep velocity gradients 10
10–2 10–1 1
provide a major source of turbulence. ε̄95%(W/kg)
One reason for widespread use of
Rushton turbines in microbial fermentors
is their ability to handle high gas-flow pitched-blade impeller is about 1. Thus, and diffusion. Distribution is also called
rates and provide efficient gas dispersal. a Rushton turbine has strong form drag, “macromixing,” diffusion is known as
The power required to achieve a given generates high levels of torque, and “micromixing,” and depending on the
stirrer speed depends on frictional forces transmits more power at the same scale of fluid motion, dispersion can be
and form drag (caused by the pressure operating speed than does a pitched classified as either macro- or
difference between the front and rear of blade impeller (1). micromixing.
the impeller blades) that resists impeller Fluid Flow: Rotating impeller blades Distribution is the process by which
rotation. Friction and form drag create pump fluid in a bioreactor vessel. The fluid is transported to all regions of a
torque on the stirrer shaft. The power volumetric flow rate of fluid leaving a stirred vessel by bulk circulation
input can be determined experimentally blade is a characteristic of the impeller currents. It is an important process in
by measuring the induced torque design and varies with operating mixing and often the slowest step
(Equation 4): parameters such as stirrer speed and because fluid has a great distance to
P = 2πNi M blade size. A dimensionless flow number traverse in a long circulation path.
where P is the power, Ni is the stirrer represents the effectiveness of a Dispersion breaks up the bulk flow into
speed, and M is the torque. pumping process (Equation 6): smaller and smaller eddies, which
The power required to mix nongassed Nflow = Q/Ni Di3 facilitates rapid transfer of material
liquids depends on stirrer speed, impeller where Q is the volumetric flow rate of through a vessel. The degree of
shape and size, tank geometry, and liquid fluid leaving the impeller blade, Ni is homogeneity possible from dispersion is
density and viscosity. The relationships the stirrer speed, and Di is the impeller limited by the size of the smallest
among those variables generally is diameter. eddies formed in a particular fluid. To
represented by the dimensionless power A pitched-blade impeller produces achieve mixing on a scale smaller than
number (Equation 5): nearly three to four times the flow of a the smallest eddies, diffusion becomes
NP = P/ρNi3Di5 Rushton turbine for the same power an important process. Such micromixing
where NP is the power number, P is input. This is why the former design is occurs relatively rapidly and over
the power, Ni is the stirrer speed, and Di recognized for high pumping capacity shorter distances than are involved in
is the impeller diameter. and effectiveness of blending operations, distribution.
Consider the power number as whereas the latter are considered to have Mixing time (tm) is a useful parameter
analogous to a drag coefficient for a low pumping efficiency (1). for assessing the overall speed of mixing
stirrer system. The relationship between For mixing to be effective in stirred in stirred vessels. It is the time required
NP and Re generally is determined vessels, the velocity of fluid leaving an to achieve a given degree of homogeneity
experimentally for a range of impeller impeller must be sufficient to carry after starting with completely segregated
and tank configurations. The power material into the most remote regions of materials. Mixing time can be measured
number for different impeller designs a tank. Circulating fluid also must by injecting tracer material — commonly
becomes constant once flow is turbulent sweep the entire vessel in a reasonable acids, bases, or concentrated salt
(1). Under turbulent flow conditions in a time. Fluid flow must be turbulent for solutions — into a vessel and following its
stirred tank, the power number for a good mixing, which can be described as concentration at a fixed point in the tank.
Rushton turbine generally remains a combination of three physical When a small pulse of tracer is added
constant at 5, whereas that of an axial processes (1): distribution, dispersion, to fluid in a stirred tank that already

JUNE 2024 22(6) BioProcess International 37

Figure 8: Mixing times measured in a 400-L stirred bioreactor at 90 W/m3 with tracer
added to the system at different points (adapted from Reference 14); numbers on the thus enabling rapid distribution to an
bars show the difference in mixing time compared with a reference addition at the top entire culture. That implication has been
of the liquid surface; θ 95% = time needed to reach 95% homogeneity in the bioreactor. understood for a long time; however, it is
not general practice.
The product of mixing time and
Reference addition on liquid surface
stirrer speed (Ni · tm) is known
Trace Addition Height

commonly as dimensionless mixing time,

28% which represents the number of stirrer
rotations required to homogenize a fluid
after addition of a tracer pulse. For
34%
turbulent flow conditions in a
bioreactor, Ni · tm approaches a constant
24% value for most stirrer types, and mixing
time reduces in direct proportion to
increasing stirrer speed. Under
0 10 20 30 40 50 60 turbulent flow conditions, tm can be
Mixing Time: Θ95% (seconds) reduced by increasing Di. However,
larger-diameter impellers also require
more power consumption, so a tradeoff
contains the tracer material at a certain tm using an iodine-decolorization balance is needed when using that
concentration (Ci), the concentration technique (14). The data in Figure 7, strategy for improving mixing and
response curve in Figure 6 can be obtained using three Rushton turbines shortening mixing time.
expected. Before mixing is complete, a and three pitched-blade impellers, show Power requirements should be kept
relatively high concentration will be that at the same power input the spatial in mind for scaling up mixing systems.
detected each time the bulk flow brings decolorization patterns and speed of The power required to achieve a given
tracer to the measurement point. The decolorization are very different for mixing time for a larger bioreactor can
resulting peaks in concentration will be those agitator types. Those results be computed from Equation 7 (1):
separated by a period approximately provide experimental evidence that P 2 = P1(VL2/VL1)5/3
equal to the average time required for impellers inducing axial flow reduce tm where P 2 is the power required for
fluid to traverse one bulk circulation by about a factor of two. The reason for the larger bioreactor, VL2 is the liquid
loop, known as the circulation time (tc). that is their notably improved top-to- volume of the larger bioreactor, P1 is the
After several circulations, a desired bottom mixing due to a more widescale power required for the smaller
degree of homogeneity will be achieved. flow structure induced by the axially bioreactor, and VL1 is the liquid volume
Mixing time generally is expressed down-pumping pitched-blade impellers; of the smaller bioreactor. For example,
as the time after which the tracer Rushton turbines induce flow consider scaling up from a 1-L to a 100-L
concentration differs from its final compartmentalization, leading to less bioreactor: P 2 = P1 × (100/1)5/3 and
concentration (Cf) by <5–10% of the total efficient overall mixing and hence P 2 = P1 × 2155. Hence, the power
concentration difference (Cf – Ci). At tm, longer mixing times. required to achieve equal mixing times
the tracer concentration is relatively A striking finding from that study is in the larger bioreactor would be
steady, and the fluid composition that mixing time depends only on ~2000× that of the smaller bioreactor.
approaches uniformity. For industrial- bioreactor geometry rather than the That is much greater than would be
scale bioreactors, that takes minutes. agitation system. Different impeller feasible economically or technically,
Liquid-phase mixing is vitally configurations influence the constant illustrating why the criterion of constant
important in large-scale bioreactors factor (10.5 or 17.9) in the empirical mixing time seldom is applied in
because feed components, bases, or correlation of Ruszkowski (15). Another scaling up. Mixing time inevitably must
antifoam detergents are added, in most experiment reported by Sieblist et al. increase with scale-up.
cases, at the top surface of the fluid (12, demonstrated that mixing times Installation of multiple impellers on
13). Such added materials must be critically depend on experimental the same stirrer shaft might be perceived
distributed quickly throughout an entire conditions, particularly the location in as a simple solution to improve mixing in
culture to reach homogeneity without the bioreactor where the tracer is added a bioreactor. However, the power
causing localized concentration (14). Figure 8 shows that the effect of required increases substantially when
gradients. The homogenization process adding tracer close to the impeller is additional impellers are fitted onto the
is dominated by prevailing flow much larger than that at the reference same shaft. Moreover, impeller design
patterns, and information about global point (the liquid surface), leading to (axial or radial) and clearance between
mixing conditions in a bioreactor can be shorter mixing times. That indicates the impellers significantly affects mixing
obtained from tm measurements. importance adding substrates and performance and would need to be
Sieblist et al. reported such a study corrective agents (e.g., bases and optimized to realize an improvement in
of large-scale bioreactors, determining antifoams) to cultures close to impellers, overall efficiency.

38 BioProcess International 22(6) JUNE 2024

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Special Emphasis on Biopharmaceutical
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in Biotechnology and Bioengineering:
To share this in PDF or professionally printed form,
Bioprocesses, Bioreactors, and Control. contact Lisa Payne: [email protected],
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