Practical Method Development Strategies for the Characterization of Proteins by Intact and Subunit Reversed Phase HPLC

Published on: 

Sponsored Content

Don’t be narrow minded in your analytical method development for subunit analysis of mAbs Widen your experimental horizon for RPLC and UHPLC method robustness and transferability. Monday, Sept.14, 2020 at 11am EDT | 8am PDT | 4pm BST | 5pm CEST On demand available after final airing until Sept. 14, 2021.

Register free:

Event Overview:

Reversed-phase liquid chromatography (RPLC) and ultrahigh-pressure liquid chromatography (UHPLC) are common approaches for both characterization and purity analysis of recombinant protein therapeutics such as monoclonal antibodies (mAbs). Intact reversed-phase LC methods are high resolution, capable of separation of hydrophobic variants such as clipping and deamidation. Additionally, intact RPLC methods are relatively quick and capable of being platformed, allowing for method implementation early in development.

However, method parameters for intact reversed-phase LC development are often undefined. Further, with the numerous approaches in generating subunits for mAbs, understanding which parameters to adjust and when can simplify what can be an overly complex design of experiment.

In this discussion, we explore experimental design space for purity analysis of intact proteins. We investigate the use of wide pore core-shell particles, and the critical role they play in gradient slope. Finally, we assess best practices in developing a robust and transferable method to support biotherapeutics development.

Key Learning Objectives:

  • Different approaches to mAb subunit analysis, and RPLC method development parameters to consider for each
  • How gradient slope and temperature play critical roles in RPLC method robustness
  • Column selection criteria for intact reversed-phase LC methods


Brian Rivera, Senior Product Manager- Biologics, Phenomenex

Time and Date: Monday, September 14, 2020 at 11am EDT | 8am PDT | 4pm BST | 5pm CEST

On demand available after final airing until Sept. 14, 2021.

Sponsor: Phenomenex

Register free: