low molecular weight peptides and spin column purification
James Ghadiali
Monday, 03 March 2008 19:10 UTC
Has anyone had any luck desalting low molecular weight peptides (~2000Da) using spin columns with LMWCO gel filtration media like Sephadex G-10?
HPLC purification is posing some problems and we would rather avoid dialysis…
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Replies
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Depends on from what else you are trying to separate it?
I routinely purify a peptide of (1900Da)with Sephadex G-25 spin column (made in-house) Separation profile is like this
a) Impurities of very low molecular weight (400-600 Da) and I can totally get rid-off
b) One of the impurities is around (~1100Da) and it always comes along with my (1900Da) product
c) Peptide recovery efficiency about 60-80 % if I try to increase the recovery I start observing increase in (1100Da) impurity.With these observation I guess you can use Sephadex G-10 for your protocol.
Hope this help.
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That’s really helpful, thanks for the info
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Anonymous
Would someone be so kind as to post their protocol for making in-house sephadex spin columns?
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