New Applications Note - AN015 Precision and accuracy of protein sizing

New Applications Note - AN015 Precision and accuracy of protein sizing

Monday, 5th July 2010

There is an increasing need within the biopharmaceutical industry for techniques which complement or enhance currently available techniques such as dynamic light scattering (DLS), size exclusion chromatography (SEC) and transmission electron microscopy (TEM) for monitoring the amount and extent of aggregation of therapeutic proteins and monoclonal antibodies. All of the techniques previously mentioned have limitations, e.g. it is not possible to determine size for proteins without dilution, they cannot be used in conjunction with in-line systems, e.g. GE AKTA explorers and also have limited application in the analysis of membrane proteins.

This application note outlines results with a test protein, myoglobin, using a technique developed by Paraytec for determining the hydrodynamic radius of species in solution. Repeatability and intermediate precision are reported, and the key factor influencing the accuracy of the method is documented. The technique measures the extent of Taylor dispersion of a plug of sample moving through a buffer solution. The extent of dispersion correlates directly with the hydrodynamic radius of the species.

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