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HILIC or IEX better for protein digest?

http://www.chromforum.org/viewtopic.php?t=9547

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HILIC or IEX better for protein digest?

Posted: Sun Nov 16, 2008 7:48 pm
by diablo
Hi folks,

i'm trying to perform a high resolution peptide map of a protein digest (tryptic). I know RP seems to be the method of choice, but i wonder if i can get similar results with either HILIC or IEX ?

Which one is better ? :D

regards
diabloo :P

Posted: Sun Nov 16, 2008 8:35 pm
by Uwe Neue
By virtue of the retention mechanism of RP, you will get better peak capacity from RP compared to IEX. With respect to HILIC, I have seen some very good resolution maps for very polar peptides that are barely retained in RP, but I do not recall having ever seen a complete digest with HILIC.

Posted: Sun Nov 16, 2008 9:42 pm
by Kostas Petritis
I suggest that you have a look at the papers below (and selected references therein):

1)Hydrophilic interaction liquid chromatography (HILIC) in proteomics Anal Bioanal Chem (2008) 391:151–159

and

2) Mixed-mode hydrophilic interaction/cation-exchange chromatography (HILIC/CEX) of peptides and proteins J. Sep. Sci. 2008, 31, 2754 – 2773

Posted: Mon Nov 17, 2008 6:46 pm
by diablo
ok, thx for your answers ! :D
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