Simulated Moving Bed Chromatography in the Pharmaceutical Industry

Ron Bates Bristol-Myers Squibb April 19, 2004

Outline
Short Biography What is Bristol-Myers Squibb Chromatography
Batch vs continuous
HPLC, LC, SMB, P-CAC

Simulated Moving Bed Chromatography

Introduction Theory (brief) Operation Applications in the Pharmaceutical Industry

 B.S. Chemical Engineering, RPI, 1993

Ph.D. Biochemical Engineering, University of Maryland, Baltimore County, 1999
Focus: ion-exchange chromatography

Pfizer, Groton, CT, 1999-2003

Focus: small molecule chromatography, HPLC, LC, SFC, SMB, FLASH, extraction, crystallization, precipitation

Bristol-Myers Squibb, Syracuse, NY, 2003-present

Focus: protein chromatography

Bristol-Myers Squibb
Top-ten pharmaceutical company Products in numerous therapeutic areas Mental Health
Abilify

Cardiovascular & Metabolic Diseases

Pravachol, Coumadin

Headache and Migrane

Excedrin

Infectious Diseases
Reyataz, Sustiva

Oncology Erbitux, Taxol Strong pipeline focused in 10 therapeutic areas

Oncology, Cardiovascular, Infectious Diseases, Inflammation, etc.

Sites around the world

U.S. Research/Manufacturing sites
MA, NY, NJ, CT, IL, Puerto Rico

Bristol-Myers Squibb
Syracuse, NY
Clinical and Commercial Manufacturing Plant
Small-molecule pilot plants
Process development and optimization Clinical manufacturing

Penicillin-based products
Last US-based Penicillin manufacturer

Bio-synthetic products Biotechnology

Development, Manufacturing, Analytical Biosciences, Quality Control / Assurance

Bristol-Myers Squibb
Syracuse, NY - Biotechnology
Two lead protein therapeutics
Abatacept: commericial in 2005
Commercial-scale manufacturing Commercial launch out of Syracuse Facility BLA filing Dec. 2004

LEA29Y: Phase III clinical trials in 2005

Development for next generation process Clinical production in 2004

Expansion in analytical and quality groups to support processes

Batch vs. Continuous Chromatography

Batch Chromatography
Discrete starting and ending points
Example: 10 minute HPLC cycle

Concentration
0

6 time

10

12

Types: GC, HPLC, FLASH, FPLC, LC, etc. Can be run in many modes:
Linear, overloaded, frontal, etc.

Batch Chromatography
Effluent to Waste Feed Load

Effluent to Waste

Desorbent

Elution

Desorbent
(Raffinate) (To Waste)

Elution

Desorbent
(Extract) Strong Solvent
Reference: Linda Wang, Perdue University

Elution

Regeneration

Batch Chromatography
Empty zone

Continuous Chromatography
Feed is loaded onto column and product is collected continuously
Feed column

Annular (P-CAC)
Preparative continuous annular chromatography

Countercurrent
Simulated moving bed chromatography (SMB)

P-CAC

Reference: Genetic Engineering News, Oct. 1, 1999

P-CAC

Reference: Genetic Engineering News, Oct. 1, 1999

P-CAC

Reference: Genetic Engineering News, Oct. 1, 1999

P-CAC

Reference: Genetic Engineering News, Oct. 1, 1999

Simulated Moving Bed Chromatography (SMB)

What is SMB
SMB is Simulated Moving Bed Chromatography. SMB is continuous countercurrent chromatography. The feed is pumped into the system and two (or more) product streams are continuously collected. SMB has been used for the production of millions of tons of bulk commodities (p-xylene, high fructose corn syrup, etc...) for the past four decades. Due to improvements in column and equipment technology, SMB has recently been used in the pharmaceutical industry (Sandoz, SmithKline, UCB, Pfizer).
HPLC costs: $100/kg to $5000/kg SMB costs: $50/kg to $200/kg

SMB versus HPLC

Advantages of SMB: Lower solvent utilization (up to 10 times less than batch HPLC) Generally can use less expensive, larger stationary phases Able to get high recovery and high purity Sometimes better productivity Lower labor and QC costs Only partial separation of solutes is required to obtain high purity. Higher yield than batch 10% more than batch. High throughput 5 to 10 fold increase. Lower solvent consumption An order of magnitude lower. Continuous process.

Disadvantage of SMB: Binary separation only Complexity

Commercial Applications of SMB

Hydrocarbons Sugars Agrochemicals Antibiotics Peptides Chiral Drugs
Gaining tremendous momentum FDA approves of the technology Chiral resin manufacturers sell resins specifically made for SMB

Proteins? Useful as polishing step?

SEC: remove aggregated form of product

Multicomponent separations more difficult than traditional uses

8, 12, even 16 zone systems being examined

Continuous Countercurrent Chromatography

Basic Principle
Feed
stationary column

Mobile Phase
A sample is injected in the centre of a stationary column The two components move at different speeds and are separated If we now move the column from right to left, at a speed halfway between that of the solutes, they now move in different directions ...

Continuous Countercurrent Chromatography

Basic Principle
Feed column

Mobile Phase
The two solutes now move in different directions relative to a stationary observer. If the column is very long, the bands will continue to separate.

Continuous Countercurrent Chromatography

Basic Principle
Feed column

Mobile Phase
If we continue to add sample at the center, the components will continue to separate

Continuous Countercurrent Chromatography

Basic Principle
Feed column

Mobile Phase
This is clearly a continuous system, but there are problems. The column needs to be of infinite length, the actual moving of solids is very difficult and some way to introduce and remove the sample and the products are needed. We solve this by cutting the column into small segments and simulating the moving of them

Continuous Countercurrent Chromatography

Basic Principle
Feed

column

Mobile Phase

The feed and solvent inlets are now placed between the segments and are moved each time a segment is moved from one end to the other

Continuous Countercurrent Chromatography

Basic Principle
column
Feed Mobile Phase

Mobile Phase
Products are removed by bleeding off a carefully calculated flow at suitable exit points. This changes the velocity of the bands in the column and forces the products to move toward the ports

This ensures that the column segments are clean before they are moved and that the solvent can be recycled directly back through the system

True Moving Bed

Binary Separation in a True Moving Bed

Raffinate Time : t Desorbent

Feed

Extract Feed Time : t + t Extract Raffinate

Desorbent
Reference: Linda Wang, Perdue University

Binary Separation in a True Moving Bed

Extract Time : t + 2t

Feed Desorbent Raffinate Desorbent Time : t + 3t Raffinate Extract

Feed
Reference: Linda Wang, Perdue University

Binary Separation in a True Moving Bed

Raffinate Time : t + 4t Desorbent

Feed

Extract Feed Time : t + 5t Extract Raffinate

Desorbent
Reference: Linda Wang, Perdue University

TMB to SMB
Since its very difficult to move solids, true countercurrent chromatography does not exist. Instead, the bed is broken into many fractions and their movement is simulated by changing the inlet and outlet ports

Simplified SMB - 1
Solvent Feed

The system is started.....

Extract Solvent

Raffinate Feed

A frontal elution separation occurs in Section 3.

Extract

Raffinate

Simplified SMB - 2
Solvent Feed

The separation continues.....

Extract Solvent

Raffinate Feed

Eventually the front of pure product 1 reaches the outlet. It is distributed between the final Section and the product port

Extract

Raffinate

Simplified SMB - 3
Solvent Feed

Extract Solvent Feed

Raffinate

Finally, the mixed product reaches the outlet. To avoid collecting impure material, it is necessary to move the columns 1 position upstream.

Extract

Raffinate

Simplified SMB - 4
Solvent Feed

The frontal separation continues; at the same time, the slow moving product starts to separate from the tail of the mixed product band in Section 2
Extract Solvent Feed Raffinate

Eventually the fast moving product again reaches the outlet and more pure product is collected.
Extract Raffinate

Simplified SMB - 5
Solvent Feed

Extract Solvent Feed

Raffinate

When the mixed band reaches the end of Section 3 its tail has left Section 2 (if the separation has been correctly designed) and only pure product 2 remains in Section 2.

To avoid collecting impure raffinate, the columns are moved once more. Now, the pure component 2 is in Section 1.
Extract Raffinate

Simplified SMB - 6
Solvent Feed

Extract Solvent Feed

The second component is now collected at the Extract port while the separation continues in Sections Raffinate 2 and 3. The faster component reaches the Raffinate port and is again collected; note that the outlet concentrations are neither constant nor concurrent.
Raffinate

Extract

Simplified SMB - 7
Solvent Feed

Eventually, the mixed zone reaches the raffinate port and the columns are again switched.
Raffinate

Extract Solvent Feed

Switch

This simplified system is now in a steady state mode and will continue to cycle.
Extract Raffinate

 The moving of the bed is simulated by moving the points of feed and mobile phase addition, as well as the points of raffinate and extract removal while keeping the column positions fixed.
Extract
Packed Column

Mobile Phase

Time = 0

Feed Mobile Phase

Raffinate

Time = 1

Extract

Raffinate

Feed

SMB Configurations
The zones are made up of one or more columns Six-column SMB System

II

III

IV

II

III

IV

Eight-column SMB system

II

III

IV

II

III

IV

SMB Operation
t0
ELUENT EXTRACT

t0 + T / 2
ELUENT EXTRACT

Liquid

Liquid

RAFFINATE

FEED

RAFFINATE

FEED

SMB Operation
t0 + 1 T
ELUENT

t0 + 1 T + T / 2
ELUENT

EXTRACT

EXTRACT

RAFFINATE

RAFFINATE

Liquid

Liquid

FEED

FEED

Theory Governing Equations

For another day

Maybe

Theory Working Equations / Definitions

k1 = capacity factor = (tr-t0) / t0

= k2 / k1
Rs = 2* (tr1-tr2) / (w1-w2)

SMB Method Development

1. Start with linear batch experiments

2.
3. 4. 5.

Increase either mass or volume of load to overload the column

Calculate isotherm Determine resistance to mass transfer (if important) Calculate necessary flow rates

6.

Optimize (either on-the-fly or with a proven model)

Linear Chromatography
tr1 tr2
Concentration

t0
0 2 4 6 time 8 10 12

Batch Chromatography Experiments

Feed concentration
As concentrated as possible to minimize disruption to Zone III velocity Need to run batch experiments at appropriate concentrations and solvents

Desorbent composition
Solubility of products Strength
Trade-off between time and mobile phase utilization

Sorbent
Capacity, selectivity, resolving power

Feed Concentration
Feed concentration: Consider two systems
A: Concentrated feed B: Dilute feed

Run batch experiments to examine effect of concentration

Desorbent composition
Multiple trade-offs: Solubility of products and effectiveness of the solvent
Not always complimentary Often solubility dictates solvent composition

Speed
Low k = high throughput
More wear and tear on equipment Larger system needed

Large k = low throughput

Less wear and tear Smaller system acceptable

Choice of Sorbent
Capacity: higher = better? Selectivity: higher = better? Resolving power: higher Rs = better?

Linear Chromatography
tr1 tr2
Concentration

t0
0 2 4 6 time 8 10 12

Volume Overloading

Absorbance

time

Batch Chromatography to SMB Initial Operating Conditions

Determine optimal feed concentration, stationary phase and mobile phase composition (highest with lowest capacity factors) Calculate isotherm and mass transfer resistances

Either use software package or rules of thumb to generate initial SMB flow rates

Solvent Mass Balances Flow Rates

vRecycle I vD vI II vX vII III vF vIII IV vRaff

Zone velocities vI = vRecycle + vD vII = vI - vX vIII = vII + vF vRecycle = vIII - vRaff

Overall Mass Balance vD + vF = vX + vRaff

Flow rates
Commercial SMB design models available
Given batch results from 5-10 column experiments
Flow rate, feed concentrations, retention times Solubility data

Predict zone velocities, productivities Problems:

Usually assumes simple adsorption model and lumped mass transfer coefficients Often difficult to interpret overloaded chromatograms

Rules of Thumb
Educated guesses based upon batch results from linear and overloaded experiments
VII and VIII ratio (based upon retention times) VI to flush back-side of slowest component from zone I Feed concentration and flow rate based upon solubility data and solvent mass balance

Period
The period is the time a column stays in one zone also called switching time. Changing the period has the effect of changing all 4 zones simultaneously, thus either speeding up or slowing down the solutes

Example of switching time

t0
ELUENT EXTRACT
ELUENT

t0 + 1 T

EXTRACT

RAFFINATE

Liquid

Liquid

RAFFINATE

FEED

FEED

SMB Optimization
Independent variables:
Flow rates
Recycle, Desorbent, Raffinate, Extract, Feed

Period (switching time) Thats it.

Procedure:
Get the system bound, manipulate the flow rates to maximize throughput at required purity

SMB Optimization
vRecycle I vD vI II vX vII III vF vIII IV vRaff

Questions: What is the effect of increasing the Zone I flow rate?

How would one accomplish this?

Zone II? Zone III? What if the system is underutilized (i.e., more feed can be added to the system) how would one do this without affecting the other zone flow rates?

Two component SMB System

Desorbent Feed

II

III

IV

Conc.

Bed Position Extract Raffinate

SMB Optimization
vRecycle I vD vI II vX vII III vF vIII IV vRaff

Questions: Extract contains too much of the weakly adsorbed species what do you do? If situation was reversed?

Two component SMB System

Desorbent Feed

II

III

IV

Conc.

Bed Position Extract Raffinate

SMB Optimization
vRecycle I vD vI II vX vII III vF vIII IV vRaff

Questions: Extract contains too much of the weakly adsorbed species what do you do? If situation was reversed?

Two component SMB System

Desorbent Feed

II

III

IV

Conc.

Bed Position Extract Raffinate

Examples of SMB

Two component SMB System

Multi-component System
0.8 0.7 0.6 0.5 Sulfuric Acid Glucose Xylose Acetic Acid

Ci/CF,i

0.4 0.3 0.2 0.1 0 0 10 20 Time [min] 30 40

Single-component pulse data

Reference: Linda Wang, Perdue University

Multi-Component SMB System

Desorbent Feed (1, 2, 3)

Concentration

II

III

IV

Extract (2, 3)

Bed Position

Raffinate (1)

1 Fast Solute 2 Intermediate Solute 3 Slow Solute

Reference: Linda Wang, Perdue University

Complete Separation in Tandem SMB

1 Des. Ext. Feed Raf.
Sulfuric Acid Glucose Acetic Acid

C /C

F,i i

0.5

0 Des.

5 Ext.

10 Feed

15 Raf.

20

C /C

F,i i

0.5

10 Column Number

15

20

Reference: Linda Wang, Perdue University

Profiles of a Parallel SMB

1.2 1 0.8
C /C
i F,i

D1

II
E1

III

IV

V
R1

VI

D2

VII VIII IX
E2

B(o) F

B(i) R2

Sulfuric Acid Glucose Acetic Acid

0.6 0.4

0.2

0 0 5

*
10

15

*
20

Column Number

Glucose yield: 94%

Reference: Linda Wang, Perdue University

Glucose purity: 99%

Other Questions?