High‐throughput process development: Chromatography media volume definition

Abstract

Prediction of chromatographic separations in liter scale with the help of data gathered using a microliter format is challenging, as both scale effects and differences in the formats used (plates vs. packed minicolumns) need to be addressed. The use of microplates in high‐throughput process development (HTPD) studies has become a routine. While this method is typically used for investigating chromatography conditions such as salt and pH, it can also be used for screening of different chromatography media (resins). In this study, the influence of the physicochemical properties of various media on preparation of medium‐containing microplates is discussed, and examples are presented that show how these properties can influence the conclusions drawn in benchmarking studies done using this HTPD format. The results indicate that differences in chromatographic media volume between microliter sampling techniques and packed chromatography columns do exist. Regardless of the chosen chromatography media, sampling technique or volume definition method, the simplest way to address this issue is by calibrating the microliter scale with packed‐column volumes in appropriate dry weight experiments.