Design of Simulated Moving Bed Chromatography - CPAC

Simulated Moving Bed Chromatography
for Insulin Purification:
Design Methods and Simulation Tools to
Achieve High Product Purity and High
Yield
George Weeden and N-H. Linda Wang
School of Chemical Engineering
Purdue University

Overview
• Chromatography and SMB
– Principles
– Key Barriers
• Purdue SMB Technologies
– Standing Wave Design & Optimization
– VERSE Dynamic Simulations
– Versatile SMB Equipment (V SMB)
– Knowledge- Based Design Method
• Examples
– Tandem SMB for insulin purification
– Five-zone SMB to recover six sugars from biomass
hydrolysate

CONVENTIONAL CHROMATOGRAPHY
A Batch Process
Eluent
Feed
Pre-equilibration
Adsorbent
Particles
3

Chromatography-Advantages
• High selectivity – 1 in 10,000.
• Compact volume compared to extraction.
• Lower solvent usage than extraction.
• Mild operating conditions – suitable for
fragile compounds.
• Versatile mechanisms of separation –
adsorption, ion exchange, size exclusion,
etc.

Mechanisms of Adsorption

Limitations of Batch Chromatography for
Production
• Difficult to achieve both high purity
and high yield
• Low adsorbent utilization
• High solvent consumption
• Product dilution
• Not a continuous process
• High cost

Solution I- Binary Separation in a
Continuous Moving Bed

Solution II–Moving Port or SMB
Desorbent
1
ZONE I
2
Extract
8
Solute
Movement
3
ZONE IV
7
Step N
ZONE III
4
ZONE II
Raffinate
6
5
Feed

Advantages of SMB over Batch
• Only partial separation of solutes is required to
obtain high purity.
• Higher yield than batch – 10% more than batch.
• High purity achievable without sacrificing yield.
• High productivity– 5 to 10 fold increase.
• Less solvent – 5 to 10 fold less.
• Reduced environment impact.
• Reduced footprint, equipment size, and manpower.
• Continuous process-high throughput.
• Economical for large scale separations.

Commercial Applications of SMB
• Hydrocarbons – UOP (Molex, Parex)
(1970’s)
• Sugars – Tate & Lyle (1980’s)
• Chiral Drugs (1990’s)
• All binary separations of low MW
chemicals in four-zone SMB

Key Barriers in SMB
• A four-zone SMB has 9 design parameters - 2 9 to 3 9
trials.
• Many system & operating parameters to be optimized.
• Method for multi-component separation not well
developed.
• Complex transient and cyclic steady state phenomena.
• Equipment more complex than batch chromatography.
• No commercial equipment for multi-component
separation.
• Control of batch identity and residence time
• Experiments expensive and time-consuming

Purdue’s Platform SMB Technologies
1. Standing Wave Design and
Optimization.
2. VERSE simulations.
3. Versatile SMB equipment.
4. A systematic knowledge-based
method for process design and
optimization.

Conc.
Standing Wave Design: Binary, Ideal, Linear Systems
Desorbent
Feed
I II III IV
Extract
Bed Position
Raffinate
=
= u
= u
= u
F
u
Feed
I
w,slow
II
w,fast
III
w,slow
IV
w,fast
=
Sε
b
u
III
0
u
II
0
u w,i
a
= u 0
a
/ ( 1 + Pd i )
= wave velocity of component i in zone a
= L c / t s = port movement velocity
F feed = feed flow rate
u 0
a
= interstitial velocity in zone a
e b = inter-particle void fraction
P = (1 – e b ) / e b = bed phase ratio
d i = distribution coefficient of i
S = column cross-sectional area

Conc.
Standing Wave Design- Linear, Non-ideal Systems
Desorbent
Desorption wave of Low affinity
solute confined in Zone II
Feed
Adsorption wave of Low affinity
solute confined in Zone IV
Zone I Zone II Zone III Zone IV
Extract
Bed Position
Raffinate
•Ideal System: Matching wave velocities with port
velocity to ensure high purity and yield
•Non-ideal Systems: Difference in wave velocity and
port velocity allows wave focusing in each zone

Concentration
Standing Wave Design: Multi-Component System
with Mass Transfer Effects
Desorbent
Feed
(1, 2, 3)
I II III IV
*
1 Fast Solute
2 Intermediate
3 Slow Solute
Extract
(2, 3)
Bed Position
*
Raffinate
(1)
*
Focused Standing Wave
Pinched Wave

Standing Wave Design & Optimization-
Advantages
• Four zone flow rates and port velocity are
solved easily.
• Little computation time required.
• Easy extension to nonlinear, multicomponent,
multi-zone separations.
• Achieving maximum productivity and
minimum solvent consumption for a given
configuration.
• Reduce search space by five dimensions or
more for optimization.

VErsatile Reaction SEparation Simulator (VERSE)
Mechanisms
.
Fluid
Bulk
Fluid
Bulk Convection
Adsorption
Sites
Reaction
Adsorption
Film
Diffusion
Interference
Ion
Exchange
Pore
Diffusion
Sorbent
Particle
Denaturation
Size
Exclusion
Surface
Diffusion
Fluid
Solid
Phase
Reaction
Pore
Fluid

VERSE
VErsatile Reaction SEparation Simulator
• Simulation based on a detailed dynamic rate model
(transient differential mass balances)
• Thousands of equations solved at each instance
• To predict column profiles and histories
• To understand transient phenomena in batch
chromatography, carousel, and SMB
• To understand residence time and batch integrity
issues in SMB
• To reduce the number of costly experiments

Versatile SMB
To implement complex processes for multicomponent
separations
Valve Controls
Valves
Sephadex
G50 Columns
Pumps

Features of Versatile SMB
• Dedicated valves and manifolds for high purity
and high yield
• Allows zone bypass, open loop, multizone,
multisolvent ,and multicomponent separation.
• Easy column expansibility.
• Moving port chromatography.
• Online decoupled regeneration (Animation)
• Independent column switching for variable step
time

Knowledge-Based Design & Optimization
Engineering
Parameters
Mobile Phase, pH,
Stationary Phase,
Temperature
Process Design &
Optimization
VERSE
Simulator
Experimental
Verification
Batch or
V-SMB Exp
Solubility,
Capacity,
Selectivity
Elution Profile,
Purity, Yield,
Concentration,
Productivity
Elution Profile,
Purity, Yield,
Concentration,
Productivity
Pulse,
Frontal
Voidage,
Isotherm,
Mass Transfer,
Numerical
Parameters
SWD,
Scale-Up,
Optimization
Sensitivity,
Trade-off
Optimal
Design
Optimal
Process
(Hirtzko et al., 1999; Mun et al., 2005; Wu et al., 1997; Xie et al., 2002)

Novel Purdue SMB Processes
• Pilot Scale Separations
– Insulin from zinc chloride and protein
impurities
– Six sugars from biomass hydrolysate (>10
components)
• Lab Scale
– Antibiotic Drug
– Amino Acids and Derivatives
– Lactic Acid
– Paclitaxel (anticancer drug )
– Chiral Drugs

Knowledge-Based Design of
Tandem SMB for Insulin Purification

Biosynthetic Human Insulin (BHI) Process
Recombinant E. coli.
Trp proinsulin
Proinsulin
Crude insulin
Homogenization
Cleavage, oxidative sulfitolysis, folding
Enzymatic transformation
Ion-exchange
Reversed-Phase HPLC
Size exclusion chromatography (SEC)
Purified BHI
Crystallization
Isolation & purification steps take 10 to 12 weeks.

Conventional Size Exclusion Chromatography
1
0.8
Insulin
(Product)
C/CF
0.6
0.4
Proinsulin /
HMWP
Fronting
Zinc
Chloride
0.2
0
Proinsulin / HMWP
Insulin (Product)
Zinc Chloride
Animation
0 100 200 300
Volume (L)
BHI exp. HPI exp. HPI sim.
• Existing SEC batch process
ZnCl2 exp. BHI sim. ZnCl2 sim.
• Yield < 90%
• Purity >99%
• Resin throughput = 0.23 L / hr/100 L BV

Tandem SMB for Insulin Purification
ELUENT
EXTRACT ( )
ELUENT
EXTRACT ( )
FEED ( )
FEED ( )
RAFFINATE ( )
RAFFINATE ( )
HMWP
Insulin
Zinc Chloride

SMB Experimental Validation : Ring I
C/CF
1
0.8
0.6
0.4
0.2
(b) Effluent histories at the extract port
HMWP exp.
HMWP sim.
Insulin exp.
Insulin sim.
ZnCl2 exp.
ZnCl2 sim.
C/CF
1
0.8
0.6
0.4
0.2
(a) Effluent histories at the raffinate port
Insulin exp.
Insulin sim.
HMWP exp.
HMWP sim.
ZnCl2 exp.
ZnCl2 sim.
0
0 10 20 30 40 50
Switching Step
0
0 10 20 30 40 50
Switching Step
(c) Mid-step column profiles at cyclic steady state
Desorbent Extract Feed Raffinate
1.2
0.9
Zone I
Zone II
Zone III
Zone IV
Insulin exp.
Insulin sim.
C/CF
0.6
HMWP exp.
HMWP sim.
0.3
ZnCl2 exp.
ZnCl2 sim.
0
0 1 2 3 4 5 6 7 8 9 10
Column number

SMB Experimental Validation: Ring II
C/CF
1
0.8
0.6
0.4
0.2
0
(b) Effluent histories at the extract port
0 20 40 60
Switching Step
Insulin exp.
Insulin sim.
ZnCl2 exp.
ZnCl2 sim.
(c) Mid-step column profiles at cyclic steady state
Desorbent Extract Feed Raffinate
1.2
Zone I
Zone II
Zone III
Zone IV
C/CF
0.9
0.6
0.3
Insulin exp.
Insulin sim.
ZnCl2 exp.
ZnCl2 sim.
0
0 1 2 3 4 5 6 7 8 9 10
Column number

Parameter (5,000 kg insulin /
year)
Batch Xie (2002) Overall Optimized SMB
Overall Yield (%) 89 99 99
Column
Configuration
Ring A
2-2-4-2 2-2-4-2
12 in series
Ring B 2-3-3-2 2-2-2-2
Feed Flowrate (mL/min) 11 (each) 115 115
Column Length (cm)
Diameter (cm)
Ring A
13.7 9.34
15
Ring B 13.7 7.02
Ring A 45
47.9 27.2
Ring B (12 units) 58.6 42.6
Throughput (L/h/100 L BV) .23 1.19 5.36
Solv. consumption (L/kg insulin) 150 42 55.6
Equip. Cost ($/kg insulin) 35.71 29.76 29.17
Solv. Cost ($/kg insulin) 15 4.20 5.56
Resin Cost ($/kg insulin) 40.07 7.41 1.61
Total Cost ($/kg) 90.78 41.50 36.35

SEC-SMB for Insulin Purification
• Lab-scale tandem SMB designed and validated
with experiments
• 99% yield, 5X throughput, and 1/3 solvent
consumption
• Robust and optimal process-scale operation
designed to increase throughput by 20X.
• Novel strategies created to reduce protein
residence time and maintain batch identity
31

Design &
Optimization
(Mun et al., IEC Res 2003)
Startup &
Shutdown
(Xie et al.,
IEC Res 2003)
Key Issues Solved
Splitting
Strategies
(Hritzko et al., AICHE J 2002)
Experimental
Validation
(Xie et al., Biotech Prog 2002)
SMB
High Purity
High Yield
Low Cost
Robust
Operation
(Mun et al.,
IEC Res 2003)
Insulin
Aggregation
(Yu et al., JCIS 2006)
Periodic
Regeneration
(Xie et al., WCCBME 2003)
Residence
Time
(Mun et al.,
AICHE J 2003)
Versatile
SMB
(Chin & Wang,
Sep Purif Rev 2004)
Batch
Identity
(Mun et al.,
Biotech Prog 2004)

Conclusions
• Purdue SMB technologies
–Software tools for design, simulation,
and optimization.
–Versatile SMB equipment for multicomponent
separation.
• Useful for developing high purity, high
yield separations of chemicals,
biochemicals, and pharmaceuticals.

Acknowledgements
• Eli Lilly Co.
• 21 st Century Indiana Research and Technology
Fund (28)
• Department of Energy (DE-FC36-01GO11071)
• Purdue TTI-IRL (NSF IGERT 9987576)
• SWD: Z. Ma, Y. Xie, K.B. Lee
• VERSE: R. Whitely, Y. Xie
• V-SMB equipment: C. Chin
• Insulin Purification: Y. Xie, S. Mun, C. Chin