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CHEE450: Engineering Biology. IEX Chromatography. Presented by: Nikki Apostolakis Helen So Tiffany Yu. Chromatography Introduction.

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Iex chromatography

CHEE450: Engineering Biology

IEX Chromatography

Presented by:

Nikki Apostolakis

Helen So

Tiffany Yu


Chromatography introduction
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.

Anion

Exchanger

Cation

Exchanger

vs.

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.


Ion exchange principles
Ion Exchange Principles

  • There are four main steps to IEX Chromatography:

  • Equilibration

  • Load and Wash

  • Elution

  • 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 principles1
Ion Exchange Principles

Stationary

Equilibration

Load

Wash

Elution

Regeneration


Ion exchange purpose
Ion Exchange Purpose

  • Intermediate separation and purification of MABs from bulk of impurities

  • Inlet stream Composition:

  • Acetic Acid

  • Impurities

  • MAB

  • Polysorbate80

  • WFI


Design considerations
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
Chosen Design

Chromaflow Columns

  • • Easily validated

  • • Cost-effective, safe packing/unpacking

  • • Appropriatefor most chromatography

  • techniques

  • Suitable for large scale GMP production


Chosen design1
Chosen Design

Chromaflow Columns

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
IEX Column Costing

BPG BioProcess Columns


Alternatives
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
THE END

THANKS FOR LISTENING.

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