New Preparative Chromatography Learning Tool: Peptide Isolation Primer

By May 10, 2017

Practical Approaches to Peptide Isolation

As the interest in peptides as potential drug candidates increases, the need for basic information about the isolation and purification of these peptides also grows.

Whether peptides are made step-wise using solid-phase peptide synthesis, solution phase synthesis, or isolated from naturally-occurring sources, almost all crude peptide mixtures are complex and require purification. Written by Waters scientists Jo-Ann M. Jablonski and Thomas E. Wheat, our new “Practical Approaches to Peptide Isolation Primer” discusses the peptide isolation workflow, method development considerations including column selection, choice of mobile phase modifier, the use of temperature, and gradient optimization, along with other relevant topics.

The use of mass-directed isolation, which makes the purification process easier with less ambiguous discrimination between the target peptide and the contaminants, is also discussed.

“Isolation of the peptide product can be improved by applying method development principles that are often associated with analytical HPLC. The separation mechanisms used to create a purification method at the larger preparative scale are the same as those principles that are utilized at the smaller scale. Column selection, choice of mobile-phase modifier, the use of temperature, and gradient optimization are choices that must be examined in the development of any separation method. Systematic method development for peptide isolations can improve the purity and yield of product significantly,” note the authors.

Chapters include:

  • Peptide purification process requirements
  • Peptide isolation workflow
  • Purification process considerations
  • Method development considerations
    • Column selection
    • Practical peptide isolation example
  • Considerations for mass-directed purification


Categories: Technologies