Transcript Chromaflow Columns

CHEE450: Engineering Biology
IEX Chromatography
Presented by:
Nikki Apostolakis
Helen So
Tiffany Yu
Chromatography Introduction
The term “chromatography” applies to a wide range of
separation techniques that are based on the
differential interaction of molecules between a moving
phase and a stationary phase.
Column chromatography is a commonly used technique
for protein purification.
Different column chromatography methods exploit
different properties of the protein; Ion exchange
chromatography separates molecules based on charge.
Anion
Exchanger
vs.
Cation
Exchanger
Ion Exchange Principles
There are four main steps to IEX Chromatography:
o Equilibration
o Load and Wash
o Elution
o Regeneration
Resins are the medium for separation and
represent the stationary phase. The surface of
these particles provide sites for selective
binding. Separation is based on reversible
adsorption of ions onto resin
Ion Exchange Principles
Stationary
Equilibration
Load
Wash
Elution
Regeneration
Ion Exchange Purpose
Intermediate separation and purification of MABs from bulk of impurities
Inlet stream Composition:
o Acetic Acid
o Impurities
o MAB
o Polysorbate 80
o WFI
Design Considerations
COLUMN DESIGN
•
•
•
•
Flow rates
Aspect ratio (H, d)
Column material
Space constraints
COLUMN INTERNALS
• Packing material
• Buffer pH’s
• Buffer composition
Chosen Design
Chromaflow Columns
Why the Chromaflow line?
• Easily validated
• Cost-effective, safe packing/unpacking
• Appropriate for most chromatography
techniques
• Suitable for large scale GMP production
Chosen Design
Chromaflow Columns
Specifications
Resin Type: Sepharose High Performance
Resin Volume: 207.88 L
Column Type: GE Chromaflow
Column Dimensions:
Inner diameter: 1800 mm
Height: 100-300mm
Pressure Rating: 3 bar
IEX Column Costing
BPG BioProcess Columns
BPG Family
(Model #)
ID
(mm)
Bed Height
(cm)
Packing Volume
(L)
Purchase Cost
($)
BPG 100/500
100
0-26
0-2
$5,678.40
BPG 100/950
100
45-54
3.5-4.2
$6,171.90
BPG 200/500
200
0-26
0-8.2
$7,677.60
BPG200/950
200
45-54
14.1-16.9
$8,922.90
BPG 300/500
296
0-26
0-17.9
$15,682.80
BPG 300/950
296
45-54
31-37.2
$18,244.80
BPG 450/500
446
3-23
4.7-35.9
$35,631.75
BPG 450/1000
446
47-58
73.4-90.6
$43,548.75
Alternatives
CONFIGURATION
Can make batch process
continuous by running columns in
parallel
SEPARATION
Change resin to have impurities
bind and allow MAB to pass
Use a different basis for
separation (e.g. molecular size)
Downstream Processing
THE END
THANKS FOR LISTENING.
ANY QUESTIONS?