What are the factors affecting the resolution in size exclusion chromatography?

The factors that affect the resolution in size exclusion chromatography include: flow rate, column length, sample volume, and particle pore size. A higher flow rate results in a higher resolution as well as more defined chromatographic peaks as a result of the suppression of protein diffusion. High flow rate also results in incomplete separation, and causes concurrent elution of different sized proteins. It also produces spectral overlap between peaks in the chromatogram. Column length is another factor, and a larger length requires the sample to diffuse through more particles, allowing greater separation. Larger pore size will also allow larger proteins to enter, resulting in a longer elution time as well as a high resolution of larger samples compared to that of smaller pore size.