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Guide to Size Exclusion Chromatography (SEC) for HPLC & UHPLC

What Are SEC Columns?

Size Exclusion Chromatography (SEC), also known as gel filtration chromatography, is a widely used method in HPLC & UHPLC for the separation of biomolecules based on their molecular size. Unlike other chromatographic techniques, SEC does not depend on chemical interactions between the analyte and the stationary phase. Instead, separation occurs through a porous stationary phase, where molecules are sorted according to their hydrodynamic volume. This approach ensures gentle and highly reproducible separations, making SEC ideal for applications such as protein and peptide analysis, oligonucleotide characterization, and the detection of monoclonal antibody (mAb) aggregates.

How Do SEC Columns Work?

The separation process in SEC is based on molecular exclusion, meaning that smaller molecules enter the pores of the stationary phase and are retained longer, whereas larger molecules are excluded from the pores and elute faster. This mechanism allows for precise separation without unwanted secondary interactions. The method is widely used in biopharmaceutical research, protein purification, and quality control, where accurate molecular weight determination is critical.

 

Choosing the Right SEC Column for Your Application

YMC offers a selection of SEC columns tailored to different bioanalytical needs:

  • YMC-Pack Diol SEC columns: Designed for a broad range of biomolecules, from small peptides to large proteins, with multiple pore sizes available (60, 120, 200, and 300 Å) to optimize separation efficiency.

  • YMC-SEC MAB columns: Specifically developed for the analysis of monoclonal antibodies (mAbs), providing exceptional resolution across both high and low molecular weight ranges, making them particularly effective for aggregate and fragment detection.

  • Multiple column dimensions: Supporting both analytical and preparative scale separations, ensuring maximum flexibility for various SEC chromatography workflows.

Applications of SEC Chromatography in HPLC & UHPLC

ISEC is an essential technique in many bioanalytical workflows, particularly in biopharmaceutical research, where it is used for protein characterization, biosimilar development, and vaccine analysis. It is also widely applied in protein purification and quality control, ensuring batch-to-batch consistency in biotherapeutic formulations. Another key application is aggregation studies, where SEC enables the reliable detection and quantification of protein aggregates, ensuring the stability and efficacy of pharmaceutical products.

 

For expert advice on selecting the optimal SEC column for your analytical needs, contact YMC Europe today.

Contact us for consultation
  • YMC Europe GmbH
  • Phone +49 2064 427-0
  • Schöttmannshof 19
  • Fax +49 2064 427-222
  • D-46539 Dinslaken