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Abstract
The following equation was derived for the resolution, R s, of proteins in linear gradient elution ion-exchange and hydrophobic interaction chromatography: R s ∝ [(column length · molecular diffusivity)/(slope of the gradient · column gel volume · linear mobile phase velocity · particle diameter 2)] 1 2 . Experimental results obtained under a wide range of experimental conditions (column length, 1–30 cm; particle diameter, 37–92 μm; linear mobile phase velocity, 0.2–3.5 cm/min; temperature, 15–35°C) have shown that this relationship is valid except for very short columns with a shallow slope of the gradient and for low flow-rates. The relationship is very useful for scaling-up the linear gradient elution of proteins from data obtained with small columns.
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