In my Protein purification I am employing Source 15Q, anion exchange beads, to purify my protein.
(a) when I start with fresh beads, my protein is getting separated from the impurities and it is working well,
(b) when I want to do another batch of purification on the same beads my protein is not getting separated from the impurities.
Remember the beads are thoroughly done cleaning in practice (CIP) and regenerated according to the manufacturer's recommendations, but still the protein is not getting separated from the impurities.but I use fresh batch again separation is excellent. under same buffer conditions, gradient etc
Can anybody help me in sorting this problem
thanks in advance,