A question about affinity chromatography (recombinant His-tagged protein purification with an Akta system). Please suppose I use an 1 mL column (nickel), I load my clarified extract, I wash and I perform the elution with a linear imidazole gradient. I get a peak in B5 fraction (for instance..).
Now, I want to scale up : I perform the purification exactly in the same conditions but I use 5 times more extract (5L culture instead 1L before) and I set on the system a 5 mL column (exactly the same type).
The question : in which fraction will I find my peak ? B5 ?
Thank for your help in advance: