Diafilter

Anthony Holmes
Will MacMillan
Ben Thorek
Andrea Weeks

Presentation outline
Overview of diafiltration
Continuous diafiltration
theory
Design
Cost analysis
Design alternatives

www.en.wikipedia.org

Overview of diafiltration
Purpose:
Removal of small molecular
components (primarily salts)
from protein-containing
mixtures

Uses ultrafiltration
membranes
Membranes allow for
separation based on:
molecular sizes
densities
http://www.pall.com

Overview (2)
Important terminology:
Filtrate: material that passes
through the membrane
Retenate: material that
remains on the filter

Diafilter considerations:
Molecular size of particles
Select filter with MWCO

Membrane flux rate

Overview (3)
Advantages of diafiltration:
Quick, efficient process
Product loss is minimized
Traditional filters would require extra steps and
costs!
Material filtering a buffer exchange occur all done
within the unit

Common techniques:
Discontinuous (volume reduction or sequential dilution)
Continuous (less harsh and allows for better stability)

Continuous filtration
Addition of eluent into diafilter where rate
of incoming buffer solution to membrane
flux rate is 1:1 (ensures constant volume)
Involves a buffer solution wash
Direct correlation between diafiltration
volumes required to successfully filter a
solution

Continuous filtration (2)

Permeability measured by rejection
coefficient (RC)
Cpi: concentration in filtrate
Cci: concentration in retenate

To determine fraction of solute in retenate:

Design
Start with defining process goals
Final product concentration
Feed volume reduction
Extent of buffer exchange
Contaminant removal specification

Will the desired product be in the retentate

or the filrate?

J Q/ A

Design (2)
Desired product in the retentate
Select MWCO 3-5x smaller than product

Calculate Flux
J = Q/(A.t)

Filter Area Calculations

Filtrate Volume / (J x Process Time)
20% Safety factor

Design (3)
Calculate Diavolume washes
Based on product loss in filter
Product Loss = 100e(R-1)[lnVCF+N]
Where R is the rejection coefficient of the membrane
N is the number of number of diavolume washes
VCF is the concentration ratio

Design (4)
Component

Required
Parameter

Selected
Filter Specifications

Omega OS filter MWCO

<1.867 kD
MWCO

1 kD MWCO

Filter Area

3m2*

0.9-18.5m2

Working Volume

20L

10-4000L

*SuperPro calculated 0.96m2 for the filter area

Cost analysis: membrane

$200/m2 (SuperPro)
Polysulphone GR61 ultrafiltration membrane on a
polypropylene support material with MWCO of
20,000 Dalton (retains 100% of protein)
2 stage diafilter is approximately 138/m2 or $166
USD/m2 (Lipnizki and Boelsmand, 2002)
$200/m2 will be used
process only demands an area of 0.96m2
cost $192 to replace the membrane every 2000 hrs
$768/year

Cost analysis: power consumption

200W/m2 of membrane
(SuperPro)
12.9kW/51.6m2 or
250W/m2 (Lipnizki and
Boelsmand, 2002) (this
value was used)
Utilities Kingston
(Industrial Rating):
$0.59/kWh+$6.45/kW+
$195.48/month)
Power costs: $688.57
CAN/membrane lifespan
or $2754.28 CAN/year

www.dribbleglass.com

Cost analysis: apparatus

Diafiltration apparatus with a membrane area of
50m2 costs $151 282 USD (Vankova et al., 2005)
Correlates to $2904 USD for above design*

www.scilog.com

Possible Setups & Alternatives

Normal vs. tangential flow filtration:
Microfiltration
Reverse osmosis

Ultrafiltration

Tangential flow filtration

Designed for concentrating and purifying
protein
Increases economic life of membrane

Millipore, 2006

Microfiltration
Commonly used to purify
water
Filters out major pathogens
porous membrane (0.1-10m)

Why not? Because microporous membrane would not

effectively remove salts
Either flow type would
produce the same result

Reverse osmosis
Similar to nanofiltration
Very tight membranes used to separate salts
and small molecules from water
Would not work for insulin
Salts would not be separated from insulin

Could be used with either flow type

Ultrafiltration
Alternative to diafiltration
Designed for desalting and separating proteins from a
buffer solution
Membrane pore size ranges from 1kDalton to 1000kDalton
(Dalton is a molecular weight measurement that
corresponds to the MWCO of the filter)
Does not include addition of buffer buffer solution
(limitation)
Additional tank and water source would be required

References

Lipzinski F. and Boelsmand J. 2002. Concepts of industrial scale diafiltration

systems. Desalination Issue. 144: 179-184

Millipore. 2006. Protein concentration and diafiltration by tangential flow

filtration an overview. Website: http://www.millipore.com/publications.nsf/docs/tb032

Vankova K., Antosova M., and Kolapovic M. 2005. Design and economics of
industrial production of Fructosyltransferase. Chemical Paper. 59(6a): 441-448

Millipore (2003) Protein Concentration and Diafiltration by Tangential Flow Filtration

http://www.millipore.com/publications.nsf/86714eb8c10dcec485256b3c005a8bd7/ab3
ba3a9d06cc6f185256bd10068b0de/$FILE/TB032.pdf

Sevick, S. H. (2005) Compatability of Various Pharmaceutical Agents with Pall Medical

Supor Membrane Filter Devices Pall Medical, MI