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Old Dec 7th 2016, 06:01 AM   #1
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Protein separation using prep chromatography

Dear all,

Iīm currently working on a separation process for a protein. The protein is very basic with high pI-values (two forms of protein - short and full because of a specific cleavage site). For both, short and full sequence, the isoelectric point is above 10.

I tried to separate with CEX column using a salt gradient with variation in gradient length and flow rate but it didnīt work.
I am not allowed to use SEC for the separation because, later on I have to upscale my process.
I also tried some pH-gradients but the chemicals I used absorbed at a wavelength of 214 nm (sequence of my protein is without an aromatic AA so it does not absorb at 280 nm).

If you have any ideas, inspirations,... please help me

Thank u,
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