Successfully Developing and Using Size-Exclusion Chromatography for the Analysis of Biotherapeutic Proteins

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Stephan M. Koza, Pamela C. Iraneta, Priya Jayaraman, Erin E. Chambers
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Size exclusion chromatography (SEC) is a LC-based technique commonly used to monitor protein aggregates throughout the commercialization process of a biotherapeutic protein including monoclonal antibodies. Traditional HPLC-based SEC methods have used columns packed with >3 μm particles of an appropriate pore size for the separation of proteins of different hydrodynamic radii. Ideally, the proteins are separated solely on their relative size in solution with larger aggregate species eluting prior to the desire monomer drug.
This technical poster details the important instrument, method, and SEC column use considerations in order to obtain robust, high resolution, UHPLC-based SEC separations. It will also highlight ways to substantially increase SEC column life that can be adversely affected by injection of microbial and sample based particulates.


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2017hplc_jayaraman_smk_sec PDF 1263.4kB