Protein Applications for Advanced Multi-Detector Size-Exclusion Chromatography

Sep 18 2012

Author: by Mark Pothecary, Stephen Ball, Paul Clarke on behalf of Malvern Instruments Ltd

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Size-exclusion chromatography (SEC) or gel permeation chromatography (GPC) is frequently used in bioscience laboratories to characterise purified and recombinant proteins. This technique has also been used to measure molecular weight or to resolve mixtures of monomers, oligomers and higher order aggregates. Using this method, molecular weight is calculated by creating a calibration curve of standard globular proteins and comparing the elution time of an unknown to the standards. This is limited in that it is based on a stable relationship between molecular weight, size, shape or hydrodynamic volume. This relationship varies significantly between proteins introducing unquantifiable inaccuracies in all measurements made in this way.

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