Ion Exchange and Reversed Phase Chromatography as Two Orthogonal Techniques for Purification of Therapeutic Peptides

Life Sciences, Pharmaceutical, Pharma Manufacturing & Supply Chain, Laboratory Technology,
  • Tuesday, October 12, 2021

Regardless of the peptide production method (peptide synthesis or isolation from expression in host cells), all crude peptides need to be purified and the purity requirements for therapeutic peptides are particularly stringent. Reversed phase chromatography (RPC) is the most commonly and frequently used peptide separation technique but has its challenges. Impurities from the peptide feed are prone to cause fouling of the column and not all impurities can be resolved on the RPC column. To add another orthogonal step, to both protect the RPC column and to increase the resolution, could solve these issues. For example, a variety of peptide modifications that cannot be removed by RPC can be separated by ion exchange chromatography.

By introducing a cation exchange chromatography (CIEX) step upstream of the high-performance silica-based RPC step, the burden from impurities is significantly reduced. Additionally, the purity of the target peptide is greatly enhanced.

Register for this webinar to hear the speaker present data on three differently produced therapeutic peptides, two solid-phase synthesized peptides and one peptide expressed in Escherichia coli, both purified with an upstream CIEX step before one or two RPC steps. Two of the cases presented are directed more towards process scale and one is specifically directed against screening and small-scale purifications.

Speaker

http://Cecilia%20Unoson,%20Bio-works

Cecilia Unoson, Manager Applications Research/ Principal Scientist, Bio-Works (Uppsala, Sweden)

Cecilia Unoson is a principal scientist and manager for the applications research team at Bio-Works. She joined the company in the beginning of 2018. She is a molecular biologist with a PhD in microbiology (Uppsala University). Prior to her work at Bio-Works, she has experience at both smaller companies, as well as in bigger companies, and spent many years (as a PhD student, postdoc and researcher) in academia studying regulatory RNAs in bacteria, especially focusing on the CRISPR-Cas9 system.

Message Presenter

Who Should Attend?

This webinar will be of interest to those working in the biopharma industry, with relevant job titles including:

  • Process developers
  • Scientists developing peptide purification protocols
  • Chromatography specialists
  • Downstream process developers
  • Heads of process development

What You Will Learn

  • How to alleviate the problem of impurities from a peptide feed fouling RPC columns
  • How introducing a cation exchange chromatography (CIEX) step upstream of high-performance silica-based RPC significantly reduces the burden from impurities and enhances purity of the target peptide
  • How a variety of peptide modifications that cannot be removed by RPC can be separated by ion exchange chromatography
  • Examples of two solid-phase synthesized peptides and one peptide expressed in Escherichia coli, each purified with an upstream CIEX step before one or two RPC steps

Xtalks Partner

Bio-Works

Bio-Works is based in Uppsala, Sweden. We develop, produce and deliver innovative agarose-based products for purification of peptides, proteins and other biomolecules to industry and research organizations in the Life Science field. The employees in the company have extensive experience in the biotechnology industry and deep knowledge about the development of high-performance purification products. This has made it possible to develop a broad and high-performance product portfolio available in various prepacked formats as well as in several different bulk sizes.

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