Under high concentration overload conditions, resolution during size exclusion chromatography of proteins can be compromised due to nonuniform flow caused by the viscous fingering flow instability. In this work, the nonuniform flow under these chromatographic conditions is analyzed by numerical simulation and magnetic resonance imaging. On the basis of an improved understanding of the flow phenomena, a new column design is proposed. The effectiveness of the design is evaluated by chromatography, magnetic resonance imaging, and numerical simulation.