Sunday, 17 December 2017

molecular biology - What buffer should I choose for IEX chromatography for purifying IgY


I will use ion exchange chromatography with an anion exchange column to purify chicken IgY. Prior to this I did dialysis to remove salts from previous purification steps, is it possible after this step to just change buffer system?


The pI of IgY is 5.6-7.6, and so, what buffer should I use for the chromatography? I know I want the IgY to be negative, so I would want pH in start buffer to be higher than pI?


I bought a column but it has not arrived yet. Does it come with recommendations on preferred buffer system for the chromatography or can I use any one I want? And also, what buffer should I use to elute my protein? Do you have any recommendations?


In previous steps, prior to dialysis, I used phosphate buffered saline with pH 7.


Thanks in advance




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