Chromatography is the main purification unit operation for biopharmaceuticals. However, there are so many variables or parameters such as flow-rate, gradient slope and column dimension to be tuned to find the optimized operating and/or chromatography conditions. The mobile phase must also be carefully examined as the separation and/or the binding capacity is strongly influenced. For this reason fully automated (robotic) systems using a 96-well micro-plate format have been developed by many researchers as high-throughput process development methods for chromatography of proteins. However, methods for predicting the process-scale column performance based on the information obtained with the micro-plate system have not yet been fully developed.In this study, we carried out the batch adsorption experiment (IgG-protein A system) by using the 96-well micro-plate for determining the adsorption isotherm. In most cases the working liquid volume was 0.1-0.2 mL and the gel volume was 0.004-0.010 mL. The dynamic binding capacity (DBC) values were measured as a function of residence time with a small column of ca. 1 mL (5 mm id x 50 mm). For rigid and small silica based resins it was possible to pack a smaller column of ca. 0.3 mL (5 mm id x 15 mm), which shows the same DBC values with the 1 mL column.A simple method for predicting the dynamic binding capacity (DBC) was proposed, which uses a correlation between the DBC/SBC (SBC: static binding capacity) and the normalized residence time with the particle diameter and the pore diffusivity. In this method the void fraction was considered in order to use the data for scale-up studies. The experimental data were well-correlated based on this correlation equation.

Simple and fast methods for predicting the dynamic binding capacity by using 96-well format micro-plates and small chromatography columns

243rd National Meeting (2012)

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