October 12, 2021(Tue)  10:00 PM HKT

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. Additionally, the purity of the target peptide is greatly enhanced.

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.

LEARNING OBJECTIVES:

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

Speaker:

Cecilia Unoson

Manager Applications Research/ Principal Scientist,

Bio-Works (Uppsala, Sweden)

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