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Techniques & Tools Sample Preparation, Mass Spectrometry, Technology, Data Analysis, Liquid Chromatography, Gas Chromatography, Thin Layer Chromatography, Preparative/Process Chromatography

Protein Applications for Advanced Multi-Detector Size-Exclusion Chromatography

sponsored by Malvern Panalytical

Introduction

Size-exclusion chromatography (SEC) is a commonly used tool in the biological sciences. Typically, it is used to measure the molecular weight of an unknown sample by referencing the elution time of the unknown against that of standards of known molecular weight. This can be done with a single concentration detector such as UV or refractive index (RI). Advanced multi-detector SEC systems now include up to four detectors including UV, RI, light scattering and intrinsic viscosity. Light scattering detectors allow the measurement of molecular weight without the need for column calibration. Intrinsic viscosity is a measure of molecular density and enables structural changes to be assessed. A combination of light scattering and intrinsic viscosity allows size to be calculated.

Maintaining the detectors in series is essential for maintaining maximum sensitivity across all four detectors. SEC is generally used to measure pure samples.

Figure 1: The TDAmax system

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