MEMBRANE BASED BIOSEPARATION

Recovery & Isolation

DEFINITION
A membrane is a thin semi-permeable barrier which
can be used for the following types of separation
1. Particle-liquid separation
2. Particle-solute separation
3. Solute-solvent separation
4. Solute-solute separation
FACTORS UTILIZED IN MEMBRANE BASED
SEPARATION,
1. Solute size
2. Electrostatic charge
3. Diffusivity
4. Solute shape

Purification
MODE OF MEMBRANE SEPARATION
Spiral Wound
Hollow Fibre
FILTRATION MODE
MICROFILTRATION
MICROFILTRATION
Microfiltration membranes are microporous and retain
particles by a purely sieving mechanism.

Typical permeate flux values are higher than in
ultrafiltration processes even though microfiltration is
operated at much lower TMP.

A microfiltration process can be operated either in a
dead-end (normal flow) mode or cross-flow mode
APPLICATIONS OF MICROFILTRATION IN
DOWNSTREAM PROCESSING
1. Cell harvesting from bioreactors

2. Virus removal for pharmaceutical products

3. Clarification of fruit juice and beverages

4. Water purification

5. Air filtration

6. Sterilization of products
TRANSPORT EQUATION
( )
c M
V
R R
P
J
+
A
=

c
M
s
c
r
A
V
r R o = =

c
= Cake layer
For micron sized particles
|
|
.
|

\
|
|
.
|

\
|

=
2 3
1 1
180
s
d
r
c
c
EXAMPLE
Bacterial cells having 0.8 micron average diameter are
being microfiltered in the cross-flow mode using a
membrane having an area of 100 cm
2
. The steady state
cake layer formed on the membrane has a thickness of
10 microns and a porosity of 0.35. If the viscosity of the
filtrate obtained is 1.4 centipoise, predict the volumetric
permeate flux at a transmembrane pressure of 50 kPa.
When pure water (viscosity = 1 centipoise) was filtered
through the same membrane at the same
transmembrane pressure, the permeate flux obtained
was 10
-4
m/s.
SOLUTION
BATCH FILTRATION (INCOMPRESSIBLE CAKE)
dt
dV
A
J
v
1
=
( )
c M
R R
P
dt
dV
A +
A
=

1
|
.
|

\
|
=
A
V
R
o c
o

o
= mass of cake solids per volume of filtrate
( ) ( )
M A
V
o
R
P
dt
dV
A +
A
=
o
1
P
R
A
V
P V
At
M o
A
+
|
.
|

\
|
A
=
|
.
|

\
|
o
2
PA
V R
A
V
P
t
M o
A
+
|
.
|

\
|
A
=
o
2
2
y = mx+c
t=0, V=0
V= total volume of filtrate
EXAMPLE2
A broth containing the yeast was filtered using microfilter and its time
needed to collect the volume of filtrate is as below
Filtration Time (sec) Volume of filtrate (ml)
2 20
10 56
25 90
50 175
The microfilter has a total area of 9 x 10
-3
m
2
and the filtrate has a
viscosity of 1.15 cP. The pressure drop is 68,000 Pa and the feed
contains 0.01 kg dry cake per liter
Determine the specific cake and membrane resistance
ULTRAFILTRATION
ULTRAFILTRATION
UF membranes can retain macromolecular solutes.
Solute retention is mainly determined by solute
size.
Other factors such as solute-solute and solute-
membrane interactions can affect solute retention.
Ultrafiltration is used for:
Concentration of solutes
Purification of solvents
Fractionation of solutes
Clarification
ULTRAFILTRATION IN DOWNSTREAM
PROCESSING
UF is widely used for processing:
therapeutic drugs,
enzymes,
hormones,
vaccines,
blood products
antibodies.
ULTRAFILTRATION
The major areas of application are listed below:
Purification of proteins and nucleic acids
Concentration of macromolecules
Desalting, i.e. removal or salts and other low molecular
weight compounds from solution of macromolecules
Virus removal from therapeutic products

ULTRAFILTRATION
The ability of an ultrafiltration membrane to retain
macromolecules is traditionally specified in terms of
its molecular cut-off (MWCO).
A MWCO value of 10 kDa means that the
membrane can retain from a feed solution 90% of
the molecules having molecular weight of 10 kDa.
Ultrafiltration separates solutes in the molecular
weight range of 5 kDa to 500 kDa. UF membranes
have pores ranging from 1 to 20 nm in diameter.
TRANSPORT EQUATIONS
The solvent flow is proportional to the applied force
(solvent velocity) (force on solvent)

P L j
p v
A =
L
p
is the solvent permeability
Include the osmotic pressure
( ) [ A A = P L j
p v
( ) [ A A = o P L j
p v
For solute leak through the membrane
is the reflection coefficient, 0 < <1
t A
is osmotic pressure which can be determined from its solution
concentration
RTC = At
GOVERNING LAWS DARCY LAW
Darcy's law is a simple proportional relationship
between the instantaneous discharge rate
through a porous medium, the viscosity of the
fluid and the pressure drop over a given distance.



The total discharge, Q (units of volume per time, e.g., m
3
/s) is
equal to the product of the permeability of the medium, k (m
2
),
the cross-sectional area to flow, A (units of area, e.g., m
2
), and
the pressure drop (Pa), all divided by the viscosity, (Pa.s)
and the length the pressure drop is taking place over.

MODEL-POISEUILLE FLOW
The flow of a solvent through ultrafiltration
membranes can be described in terms of a pore
flow model which assumes ideal cylindrical pores
aligned normal to the membrane surface:
TRANSMEMBRANE PRESSURE
The transmembrane pressure in cross-flow UF is
given by:
CONCENTRATION POLARIZATION
the retained macromolecules accumulate near the
membrane surface caused concentration
polarization.
At steady state, a stable concentration gradient
exists near the membrane owing to back diffusion
of solute from the membrane surface.
offers extra hydraulic resistance to the flow of
solvent
development of osmotic pressure which acts
against the applied transmembrane pressure
MODEL
As most ultrafiltration membranes can not be
visualized as having parallel cylindrical pores, a
parameter, the membrane hydraulic resistance is
used for calculating permeate flux:
If the solute build-up is extensive, a gel layer may
be formed on top of the membrane
LIMITING FLUX
At lower values of transmembrane pressure, the
permeate flux increases linearly with increase in
pressure
However, as the pressure is further increased, there
is deviation from the solvent profile, this being due
to concentration polarization.
At very high transmembrane pressures, the
permeate flux usually plateaus off, clearly
suggesting the formation of a gel layer.
LIMITING FLUX
Beyond this point, increasing the transmembrane
pressure has a negligible effect on the permeate
flux
This value of permeate flux being referred to as the
limiting flux (J
lim
).

CROSSFLOW FILTRATION
The fluid in crossflow filtration flows parallel to the
membrane surface, resulting in constant permeate
flux at steady state
MODEL
CONCENTRATION POLARIZATION MODEL
At steady state, a material balance of solute
molecules in a control volume within the
concentration polarization layer yields the following
differential equation:


Integrating this with boundary conditions (C = C
w
at
x = 0; C = C
b
at x = o
b
), we get


k (= mass transfer coefficient) = D / o
b


0 = +
dx
dC
D C J C J
p v v
|
|
.
|

\
|

=
p b
p w
v
C C
C C
k J ln
CONCENTRATION POLARIZATION MODEL

For total solute rejection, i.e., when C
p
= 0, the
equation reduces to

|
|
.
|

\
|
=
b
w
v
C
C
k J ln
ULTRAFILTRATION-EXAMPLE 1
A protein solution (concentration = 4.4 g/1) is being
ultrafiltered using a spiral wound membrane module,
which totally retains the protein. At a certain
transmembrane pressure the permeate flux is 1.3 x
10
-5
m/s. The diffusivity of the protein is 9.5 x 10
-11

m
2
/s while the wall concentration at this operating
condition is estimated to be 10 g/1. Predict the
thickness of the boundary layer. If the permeate flux
is increased to 2.6 x 10
-5
m/s while maintaining the
same hydrodynamic conditions within the membrane
module, what is the new wall concentration?
ULTRAFILTRATION-MODEL-SOLUTE MASS
TRANSFER COEFFICIENT
The solute mass transfer coefficient (k) is a
measure of the hydrodynamic conditions within a
membrane module.
The mass transfer coefficient can be estimated from
correlations involving the:
Sherwood number (Sh),
Reynolds number (Re), and
Schmidt number (Sc):

These correlations are based on heat and mass
transfer analogy.
In the case of fully developed laminar flow, the
Graetz-Leveque correlation can be used:


For turbulent flow (i.e. Re > 2000), the Dittus-
Boelter correlation can be used:
MEMBRANE PERFORMANCE - RETENTION
If a solute is not totally retained (or rejected), the
amount of solute going through the membrane can
be quantified in terms of the membrane intrinsic
rejection coefficient (R,) or intrinsic sieving
coefficient (S,):


More practical parameters such as the apparent
rejection coefficient (Ra) or the apparent sieving
coefficient (Sa) are frequently preferred:
A correlation between the intrinsic sieving coefficient and
the apparent sieving coefficient can be obtained as:



If the intrinsic sieving coefficient could be considered a
constant, above equation provides a way by which the
mass transfer coefficient and the intrinsic sieving
coefficient could be determined by plotting experimental
data

EXAMPLE
The intrinsic and apparent rejection coefficients for
a solute in an ultrafiltration process were found to
be 0.95 and 0.63 respectively at a permeate flux
value of 6 x 10
-3
cm/s. What is the solute mass
transfer coefficient?
MEMBRANE PERFORMANCE - FLUX
The permeate flux in an ultrafiltration process
determines its productivity
The permeate flux depends on:
properties of the membrane
Properties of the feed solution.
transmembrane pressure
the solute mass transfer coefficient (which affects the
concentration polarization).
membrane fouling
ENHANCING PERMEATE FLUX
By increasing the cross-flow rate
By creating pulsatile or oscillatory flow on the feed
side
By back flushing the membrane
By creating turbulence on the feed side using
inserts and baffles
By sparging gas bubbles into the feed
MEMBRANE PERFORMANCE - RETENTION
The retention of a solute by a membrane primarily
depends of on
the solute diameter to pore diameter ratio.
the solute shape,
solute charge,
solute compressibility,
solute-membrane interactions (which depend on the
solution conditions)
operating conditions (such as cross-flow velocity and
transmembrane pressure).
NANOFILTRATION
NANOFILTRATION
Nanofiltration (NF) membranes allow salts and
other small molecules to pass through but retain
larger molecules such as peptides, hormones and
sugars.
The transmembrane pressure in NF ranges from 40
to 200 psig.
GOVERNING LAWS - FICKS 1
ST
LAW
Fick's first law relates the diffusive flux to the
concentration field, by postulating that the flux goes
from regions of high concentration to regions of low
concentration,
where

J is the diffusion flux in dimensions of [(amount of substance)
length
2
time
1
]
D is the diffusion coefficient or diffusivity in dimensions of
[length
2
time
1
] (for ideal mixtures)
is the concentration in dimensions of [(amount of substance)
length
3
],
x is the position [length],

GOVERNING LAWS - FICKS 2
ND
LAW
Fick's second law predicts how diffusion causes
the concentration field to change with time (derived
from Fick's First law and the mass balance):

Where
is the concentration in dimensions of [(amount of substance)
length
3
],
t is time [s]
D is the diffusion coefficient in dimensions of [length
2
time
1
],
x is the position [length]


The extended Nernst-Planck equation proposed by
Schlogl and Dresner forms the basis of the
description of ion transport through the membranes.
The equation can be expressed as
REVERSE OSMOSIS
Reverse osmosis (RO) membranes allow water to
go through but retain all dissolved species present
in the feed.
In osmosis water travels from the lower solute
concentration side to the higher solute
concentration side of the membrane.
In RO the reverse takes place due to the
application of transmembrane pressure.
The normal transmembrane pressure range in RO
is from 200 to 300 psig.
ASSIGNMENT 3
A protein solution at a concentration of 0.5 g/liter and
volume of 1000 liters must be concentrated on a
crossflow ultrafilter to a concentration of 10 g/L. The
ultrafiltration has an area of 100 m
2
and operates at 5
o
C
with an inlet pressure of 14.0 bar, and back pressure on
the permeate of 1.4 bar. The protein has a molecular
weight of 20,000 Da and will not pass through the
ultrafiltration membranes. The feed has a viscosity of
1.2 cp. The membrane resistance (R
m
) of the ultrafilter
has been determined to be 2 x 10
13
cm
-1
. The Reynolds
number for flow within the ultrafilter is large enough to
render concentration polarization negligible. Determine
the time required to perform the ultrafiltration.
SOLUTION
( )
cp M
V
R R
P
J
+
A A
=

t o
RTC = At