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Ion Exchange Chromatography

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

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Ion Exchange Chromatography

Postby Elinor » Fri Oct 03, 2008 3:03 am

Hi everyone,
I have a question about ion exchange chromatography: When using the ion exchange column in this method, why is it necessary to ensure that the liquid does not drain below the surface of the resin in the column?
I cant find the answer anywhere. Does it have anything to do with disturbing the resin?.......Thanks
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Re: Ion Exchange Chromatography

Postby blcr11 » Sat Oct 04, 2008 12:33 pm

I don't know why it should be worse for ion exchange than for any other kind of column chromatography. If you let the fluid level drop below the top boundary, you will have to re-suspend the dry portion in order to continue. At best you won't have much noticeable effect on the eluted bands, but you may be mixing up and undoing some of the resolution you'd hoped for, and in the worst case you will lose all resolution and maybe even your sample if it sticks to the column and won't come off. Since the protein is bound to the resin after loading an ion exchange resin, I wouldn't think the elution would be that badly affected by a slight drying out of the top of the resin. Still I wouldn't recommend it, either. You may also increase the chances that the protein will stick irreversibly to the resin if it dries out with the protein bound to it. That can happen anyway even without drying, but why tempt fate.
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Re: Ion Exchange Chromatography

Postby Elinor » Sat Oct 04, 2008 4:14 pm

Ok...all that makes alot of sense...thank you!!
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