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Size Exclusion Chromatography (SEC) is a common technique in biochemistry laboratories and routinely used for purification, identification, and quantification of protein mixtures. Dynamic light scattering (DLS) is traditionally used for pre-column size & polydispersity measurements, along with aggregate quantification.

Aside from the enhanced sensitivity, a big advantage of DLS coupled with SEC is the ability to perform Absolute SEC (ASEC), or size exclusion chromatography in the absence of costly and time consuming column calibration. DLS provides a direct measure of the protein size, as opposed to the SEC derived size extracted from a calibration curve. In addition, size vs. mass relationships for globular proteins and polymeric systems are well known, and can be used to calculate the molecular mass from the DLS measured hydrodynamic radius.

• Samples are measured directly after elution from a chromatography column
• The scattering intensity, hydrodynamic size, and molecular weight of the protein are displayed in real time, under flow
• Absolute method requires no column calibration or secondary detectors for molecular weight determination
• Size distribution resolution is enhanced through combination with chromatography

For More Information on the Absolute Size Exclusion Chromatography from Malvern Instruments - CLICK HERE...

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For specific information use the links below for more information:

• Protein characterization overview
• Solutions for protein characterization applications
• Malvern equipment for protein characterization and automation
• Protein solutions for batch and SEC
• What is the hydrodynamic radius
• Malvern Zetasizer Nano for proteins
  

Case studies :

• Size examples for common proteins
• Protein sensitivity calculator
• Combining chromatography and DLS

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Malvern is a leading supplier of particle analysis solutions for characterization and rheology applications. Advanced measurement technologies are combined with robust mechanical designs, and comprehensive data handling and automation software, to provide industrially relevant systems to sectors as diverse as pharmaceuticals and cement.

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An established world leader in particle analysis systems, Malvern recently added rheological property measurement to its portfolio. Material characterization data such as particle size distribution, particle shape, zeta potential, molecular weight, and bulk material properties, can be accurately and reproducibly measured using instruments from the Malvern range. Technologies used include laser diffraction, image analysis, laser Doppler electrophoresis, static and dynamic light scattering, capillary rheometry and strain controlled and stress controlled rheometry.

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