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The CHROMacademy Essential Guide Webcast:
Method Development for Ion Exchange Using Dynamic Buffer Generation

Thursday 16th November 2017,
11:00am EST/ 8:00am PST/ 4:00pm GMT/ 5:00pm CET




This webcast will focus on understanding the complexity of biotherapeutic protein charge variant analysis and how optimization of weak cation exchange chromatography is a necessity. Development of robust experimental method conditions can only be found by a systematic screening approach, since each protein will retained differently depending on its sequence and quaternary structure. Coupling the optimized method to a Mass Spectrometry detector can be achieved by using offline or online (2D) desalting techniques to enable each peak to be fully characterized.

Presented by

Presented by Dr. Andrew Coffey (Senior Application Chemist, Agilent Technologies).

Topics Covered

  • Charge variants and how they arise
  • Use of cation exchange chromatography to analyze native protein charge variants
  • Use of software to dynamically create changing buffer conditions for method optimization
  • Transferring to MS-compatible mobile phase for greater understanding of individual charge variants

Presenter Information »


Method Development for Ion Exchange Using Dynamic Buffer Generation

This month’s webcast focused on developing ion exchange methods using dynamic buffer generation.  The CHROMacademy material provided in this month’s tutorial will give you a fuller understanding of the information which can be obtained from IEX analysis of biomolecules and how to develop these methods.

Our Quick guides, CHROMacademy course content, and archived webcasts and tutorials will provide information on the science behind ion exchange chromatography and practical information on starting conditions for method development.

Watch the webcast

eLearning Modules

Ion Exchange Techniques

This module covers the use of ion exchange for the analysis of biomolecules.  The ion exchange mechanism, elution considerations (salt vs. pH gradient), and method development are all discussed.   

Ion Exchange Chromatography for the Analysis of Biomolecules | Salt Gradient Elution Conditions
pH Gradient Elution Conditions | Column Selection for Biomolecule Analysis by IEX
Introduction to Ion Chromatography

An introduction to the chemistry behind ion chromatography.  Acid-base chemistry, ion exchange mechanisms, detection, and hyphenated ion chromatography amongst other topics are detailed.

Types of Ion Chromatography | General Ion Exchange Considerations
Ion Exchange – Mechanisms | Ion Exchange Elution Considerations

Quick Guides

HILIC, IEX, and SEC for the Analysis of Biomolecules

A step by step guide to selecting an HPLC column.

HILIC, IEX, and SEC for the Analysis of Biomolecules »


Selecting Columns and Initial Conditions for IEX and SEC Biomolecule Analysis

A step by step guide to selecting an HPLC column.

Selecting Columns and Initial Conditions for IEX and SEC Biomolecule Analysis »

Techniques Employed in Biopharmaceutical Analysis - Part II Ion Exchange and SEC

Biopharmaceuticals offer great new possibilities and potential in treating various life threatening diseases such as cancer and autoimmune diseases. However, their complexity, resulting from their heterogeneity, represents a great challenge to the analytical chemist and a suite of techniques is required in the characterization and analysis. In conjunction with RP and HILIC, covered in Part I, ion exchange chromatography (IEX) and size exclusion chromatography (SEC) are the main stay of the biopharmaceutical analyst’s arsenal. These techniques provide much more information than merely charge variance and aggregation analysis. In this event we will discuss how, why and where IEX and SEC are typically employed in biopharmaceutical characterization and quantification. We will also highlight strategies for developing more robust IEX and SEC methods such as reducing SEC secondary interactions and contrast the use of pH and salt gradients in IEX. We will also explore the affect temperature and pressure have on SEC separations and the role of UHPLC.

Ion Exchange and SEC »

HPLC Techniques in Biopharmaceutical Analysis

Biopharmaceuticals offer great hope in treating medical conditions which are currently poorly served at best by traditional pharmaceuticals.  It is estimated that there are over 400 biopharmaceuticals in clinical trials for in excess of 200 disease areas.  The enhanced complexity and variability that comes from the size of biopharmaceuticals, allied with the intricacy of the production process, mean chromatography is employed to a much greater extent during production and release testing.  This Essential Guide webcast will provide details of why differing modes of chromatography are required in the research, preparation and analysis of biopharmaceuticals and when they would be typically used.

HPLC Techniques in Biopharmaceutical Analysis »

Introduction to Ion Chromatography

A definitive guide to the fundamental theory and instrumentation of ion chromatography. This webcast considers the underlying theory of the technique, column and eluent selection and key separation variables as well as the various techniques used for ion suppression and detection. We also consider various applications of ion chromatography together with recent advances in the technique including capillary ion chromatography, fast and ultra-high pressure ion chromatography as well as hyphenation of ion chromatography to mass spectrometric detectors.

Introduction to Ion Chromatography »

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Dr Andrew Coffey

Dr Andrew Coffey
Senior Applications Chemist
Agilent Technologies

Andrew joined Polymer Laboratories in 1989 after completing his PhD in peptide synthesis at Wolverhampton, UK.

He progressed from Senior Scientist to Product Manager looking after the HPLC column product line before focusing on Solid Phase specialty particle businesses.

He continued as Product Manager adding Biosolutions and Flash chromatography consumables product lines to his portfolio during the acquisition by Varian (2005) and then Agilent (2009), and joined the Biocolumns Applications Team in August 2011.