Influence of size of RPC column on resolution, binding capacity - (May/29/2008 )
The sample - proteins from ion exchange chromatography fraction should be further purified on a C4 column for reverse phase chromatography. We want to use the column from Waters company: Symmetry300A, C4, particle size 5um. However, there are column with different size (lenght, diameter), e.g., 4.6 x 150mm or 3.9 x 150mm or 4.6 x 50mm.
What is practical consequence(s) of different size of RPC columns on protein separation (resolution) and binding capacity? For example, the column of 4.6 x 150mm has higher binding capacity (can bind more ug or mg of protein) than the same column, but with size of 3.9 x 150mm? The column of 4.6 x 50mm gives worse resolution than column of 4.6 x 150mm? Is it right?
Thanks for your suggestions.
the same packing in different size columns will be for capacity.
resolution is controlled more by the elution (gradient or strength of isocratic medium).
capacity is involved in resolution inasmuch as overloading will reduce resolution.