Scaling up of medium-performance gel filtration chromatography of proteins.

概要

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Since gel filtration chromatography with particles having a relatively small diameter and high mechanical stability (medium-performance gel filtration chromatography, MPGFC) is useful not only in scaling up the separation of proteins buj also in reducing the separation time, HETP of proteins and pressure drop were measured for MPGFC (particle diameter, 44μm) columns of various column dimensions. The smallest column was 1.0cm in diameter and 30cm in length (24cm3), while the largest was 9 × 90cm (5726cm3). The dependences of HETP and of pressure drop per unit length on column dimensions were found to be negligible for these columns. The sample volume dependence of HETP was predicted well on the basis of the HETP at small sample volumes, when the protein concentration is low. However, when the protein concentration was increased to above 3%, HETP increased sharply. This increase of HETP was considered to be due to the relative difference between the viscosity of the sample and that of the elution buffer. From the above results, we expect that the scaling up of MPGFC of proteins can be easily carried out on the basis of the data obtained with a small column.
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