1 / 11

IEX Chromatography

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

lieu
Download Presentation

IEX Chromatography

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

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

  2. 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.

  3. 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

  4. Ion Exchange Principles Stationary Equilibration Load Wash Elution Regeneration

  5. Ion Exchange Purpose • Intermediate separation and purification of MABs from bulk of impurities • Inlet stream Composition: • Acetic Acid • Impurities • MAB • Polysorbate80 • WFI

  6. Design Considerations • COLUMN DESIGN • Flow rates • Aspect ratio (H, d) • Column material • Space constraints • COLUMN INTERNALS • Packing material • Buffer pH’s • Buffer composition

  7. Chosen Design Chromaflow Columns • • Easily validated • • Cost-effective, safe packing/unpacking • • Appropriatefor most chromatography • techniques • Suitable for large scale GMP production

  8. 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

  9. IEX Column Costing BPG BioProcess Columns

  10. 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

  11. THE END THANKS FOR LISTENING. ANY QUESTIONS?

More Related