SEC overview of the principles and techniques to obtain reliable protein size-based-separations

by | May 11, 2022

Learn about the fundamentals and best practices of SEC, from the column perspective, through this open-access liquid chromatography training course by Waters.

Size-Exclusion Chromatography (SEC) is a powerful technique for protein characterization that requires an understanding of the principles to be successful. In this open-access course, we will discuss the history and science of SEC, its strengths and limitations, and how to get the most from this important analytical technique. We’ll also discuss the analytical challenges with a focus on monoclonal antibody separations and the effective use of LC-based, size-exclusion chromatography to obtain reliable mAb aggregate, monomer, and fragment analyses.


The full "Size exclusion chromatography overview of the principles and techniques to obtain reliable protein size-based-separations" open-access course includes six tracks:

  • Analytical Challenges and Use of SEC for mAbs
  • Principles and Practice of SEC for Reliable Monoclonal Antibody Analyses
  • Selecting Appropriate SEC Pore Size for Reliable Protein / Peptide Separations
  • Choosing Appropriate SEC Column Size based on LC System and Application Goals
  • Considerations to Maximize SEC Column Life
  • Developing Robust SEC Methods for Protein Size Variants

Once you complete all the tutorials in this module, including answering all the quiz questions correctly, you will receive a certificate of completion.

The Presenter

Bill Warren (Principal Product Manager, Waters)

Bill received his Master's in Microbiology and Immunology from the University of Louisville School of Medicine. In 1986, joined Waters Corporation as a senior applications chemist involved with the research and development of HPLC, FPLC, and Capillary Electrophoresis applications for the analysis and purification of proteins, peptides, and nucleic acids. Bill currently is a Principal Bioseparation Columns Product Manager whose responsibilities involve assisting in the development and commercialization of innovative reagents and chromatography-based technologies for the separation, purification, and characterization of biological molecules.

LC Tutorials by Separation Science

Now that you’ve learned about Size-Exclusion Chromatography (SEC) for protein characterization, be sure to check out our other modules:

Understanding LC Column Attributes - What's Important for Your Chromatography?

Have you ever wondered how the LC column creates a separation? What attributes are important, what do they do, and how to use them to your advantage? This open-access course is designed to take you from beginner to advanced in understanding column selection and method development. It will cover the key aspects of the LC column from the base particle to the bonded phase and even the hardware.

How to improve method robustness through control of unwanted interactions

Successful chromatographic separations are as much about preventing the interactions you don’t want, as it is about creating the interactions you need. This open-access course is designed to take you from beginner to advanced in controlling the interactions within your chromatographic separations. It will cover how to improve method robustness through the prevention of analyte loss, with improved confidence in the quality of your data.

Understanding the role of SPE for cleanup of biological matrices in bioanalysis

This module series will introduce all common types of sample preparation used in the bioanalytical laboratory space and then will focus primarily on solid-phase extraction (SPE) which yields the highest cleanliness of sample extracts. Gaining a fundamental understanding of how SPE works, what benefits SPE provides, and what SPE devices are available in the laboratory will be a main focus, from learning about reversed-phase and mixed-mode ion exchange retention mechanisms, to selecting a sorbent, device format, and processing method.

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