Interlaced Size Exclusion Chromatography for faster protein analysis

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Abstract

In this research note, interlaced Size Exclusion Chromatography (SEC) is used to shorten the duration of protein analysis. The applied method has a threefold higher sample throughput than comparable ones using conventional SEC. Further important advantages are the identical area’s precision and the preservation of protein stability. Two different applications are presented where a short time interval of analysis is very important. One application is the determination of aggregates of monoclonal antibodies (mAbs). A high number of antibody samples have to be analyzed during the development process. The antibody sample was measured in a series of 60 injections within only 5.5 h instead of 16 h. The second application is the two-dimensional (2 D) separation of protein mixtures. The number of samples typically increases from the first to the second dimension. Thus, interlaced SEC is a reasonable option for the second dimension. This work confirms the functionality of interlaced SEC and widens the applicable range. Interlaced SEC can become a convenient option for method acceleration in the future.

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