The amazing ability of continuous
chromatography to adapt to a
moving environment.
Roger-Marc Nicoud,
Founder of Novasep
Barcelona, October 2013
Introduction
My thesis:
the environment pressure pushes Continuous
Chromatography to evolve like animals evolved
according to Darwin’s law ….
and for that, I will use shortcuts , approximations and manipulations …
Improving what ?
Counter-current can help
Continuous can help
System size
Eluent
consumption
Yield
Operations
Comment
Productivity
kg/kg/day normally
presented as the
main objective
function
Minimizing solvent
consumption (prior
to recycling) is a
must in all large scale
applications.
Can be a major
cost contributor
Control / Process
Stability / Cleaning can
be critical costs
contributors.
Mitigation
large LP systems can
be cheaper than
smaller HP systems
Biopharma industry
still a bit less
concerned
Ancillary equipment is
even more important
than chromatography
itself.
Life is often a matter of compromize
Continuous is NOT synonymous with
counter-current
A continuous but NOT counter-current animal
Be careful with comparisons
Productivity kg/kg/d
12
10
8
Optimum particle size, eluent
composition … are different for Batch
and continuous/CC processes.
6
4
2
0
Batch
My clever
process
S.M.B.: the key evolution …..
Evolution lead to the first continuous AND counter-current animal
solid
Maximize productivity
liquid
IV
III
Feed
II
I
Eluent
The first evidence of continuous
chromatography
Environment pressure : Paraxylene
• Scale (500,000 Mt/Y/Unit in 2010)
• Chromatographic conditions:
– Selectivity about 2
– Particle size about 1000 microns (inducing “poor” HETP ….)
– Temperature about 180 °C (inducing low viscosity)
• Technical constraints
– Target purity: > 99.8 %
– “individual bed length” : limited to 1 m due to mechanical constraints
significant bed length of 24 m
(no pb because of low viscosity)
24 columns to reach total length
Environment pressure : Fructose
• Scale (50,000 Mt/Y/Unit in 2010): from Glucose-Fructose
mixtures
• Chromatographic conditions:
– Selectivity about 1.5
– Particle size about 300-500 microns (inducing “poor” HETP)
– Temperature: 60 °C (thus viscosity about 10 cp )
• Technical constraints
– Target purity: about 90 %
– Minimize energy consumption for evaporating Water
total bed length of about 8-12 meters
Improving the classical SMB
• Add some degree of freedom by:
– Connecting columns in different ways
– Selecting different switching modes
– Varying I/O lines composition and flows
• New processes are normally:
– quasi-continuous instead of being truly continuous
– taking advantage of «counter-current» contact
Fructose: further improvement
A new animal
Two sub periods
«Cousin animals»:
• SSMB Novasep
• ISMB Mitsubishi
Commercial solutions implemented:
• 4 or 6 columns
• Eluent consumption decreased by about 10-15 % compared to SMB
Environment pressure : optical isomers
• Scale (1-100 Mt/Y/Unit in 2010)
• Chromatographic conditions:
– Selectivity: typically between 1.2 and 2
– Little influence of particle size on packing cost
– HETP depending on particle size …
• Technical constraints
– Target purity: 95-99 %
– Minimize system size / CSP inventory
Use small particle size
Total bed length about 0.5 m ….
To be distributed between Ncol …
At large scale L/D < 0.1 …..
The first SMB animal for separating optical isomers (1992):
Influence of petrochemical design is very visible ….
and the modern version ….
Optical isomers: further improvement
VariCol
VariCol:
• Truly continuous
• Same hardware as SMB
• Typically saves one
column out of 5-6.
Key successes
For main successes, continuous
chromatography is coupled with
crystallization, isomerization and
multiple effect evaporators
Very large scale does not mean
« second-class » performance
In Line Dilution is key
Environment pressure : need for
multicomponent (bio)separations.
The generic purification problem :
An animal with no progeny ?
VIII
VII
Technically one can design
a continuous three pure
fractions TMB.
VI
Prefer two SMB in series :
Paclitaxel, Cyclosporine,
EPA …
W P
S time
reduces to a binary separation if one wants
or change
…
the green
or the concept
red product.
SMB is perfectly adapted to these situations
(ex.: desalting, xylenes, capture …),
V
IV
III
II
I
Imposes to work with two
suboptimal systems ?
Improving multicomponent separations
MCSGP (Multicolumn Countercurrent
Solvent Gradient Purification) from ETH.
VI
P+W
V
W
W
•
•
•
Feed
P
IV
S
P+S
P
III
II
S
I
Important features of the animal:
Opening liquid loop
Short-circuit
Gradient
A truly continuous gradient
chromatographic process with 6-columns
3-column MCSGP
Two sub periods
VI
P+W
VV
VI
W
P+W
V
“Cousin animal” from Novasep : GSSR
IV
IV
Gradient (with) Steady-State Recycling
P+S
III
P
II
P+S
III
II
S
I
I
Can be
done with
three
columns
Not strictly
continuous
but counter
current
Environment pressure :
need for capturing (biomolecules)
Selectivity is such
that a solvent
change is
mandatory
t
Loading
Washing
Elution
W
Non
retained
P
S
Very strongly
retained
tcycle
Regeneration
Equilibration
Bioseparations: the BioSc concept
Two sub periods
W
Feed
Wash
W
Wash *
W animals” from :
“Cousin
•
•
Chromacon (Capture SMB)
GE (3C PCC) Wash
P
Target
Elute
P
Target
Elute
S
Regenerate
S
Regenerate
Waste
Equilibrate
W
Waste
Equilibrate
Use sequential
feeding to reduce
the number
columns (as low as
two).
Lysine production main steps
FERMENTATION
MEMBRANE
FILTRATION
BIOMASS
ION EXCHANGE
PURIFICATION
About 50,000 MT/year/unit
ACIDIFICATION
EVAPORATION
CRISTALLISATION
CRUDE L LYSINE HCl
Lysine purification: switch from batch to continous
(a decade ago)
• First step: Strong Cation Exchange
• Feed pH 3-4 so that Lys+ (maximizes loading and separation)
• Then pH lowered to 1-2 so that Lys++ (maximizes capture of small
concentrations)
• Wash
• Elution by ammonia at pH 9 (Lys0)
• Ammonia recycled through stripping
 97% Lys with microbial byproducts and mineral cations
• Second step: Weak Cation Exchange
• Feed at pH 9 (Lys0 goes through)
• Regeneration at pH with diluted sulphuric acid
• Wash
Gain continuous (about 10-column systems) vs Batch:
resin inventory and effluent divided by about 3
Biopharma
Many proof of concept published:
• mAbs
• pegylated proteins
• peptides
• fatty acids …..
Still looking for a
first Biopharma
industrial
application of
continuous
chromatography ?
Evolution ….
Continuous character
Xylene
100 %
Number of columns
24
Sugars
12
I-SMB
S-SMB
6
VariCol
MCSGP
4
1960
1980
2000
MCSGP
GSSR
BioSc
3C PCC
50 %
2
2010
2020 ?
Thank you !
Process integration may lead to
non intuitive processes …
Intuition
Glu 90 %
Glu 95 %
Isomerization
Glu-Fru 58-42
S
M
B
HFS 55 %
Optimized
Glu 90 %
Glu 95 %
Isomerization
Glu-Fru 58-42
S
M
B
Fru 90 %
HFS 55 %