synthetic peptide purification , help

hanieh ariani · #1 (**permalink**) 12-22-2008, 05:38 PM

hi
my peptide has 18 aa with 50% hydrophobicity, i do't know why my c18 RP- column can not purify it. all peaks are ligated to each other . what i have to do?

glbiochem · #2 (**permalink**) 12-23-2008, 02:14 AM

whether your sequence has aggregation part?

hanieh ariani · #3 (**permalink**) 12-23-2008, 06:33 AM

hello
how i can recognize that it has been aggrigated or not?
the sequence: GLAANCKRTIKCFLPKIF
thanks for your attention

glbiochem · #4 (**permalink**) 12-25-2008, 05:18 AM

It seems that the peptide's N terminus is some aggrigation.
I usually use "peptide companion" to predict it. However, I'm a sales person, not technician. The predict result is not always consistent with the real result.
I just use this software result to provide some suggestion for my customer.

Regarding to your sequence, maybe you can consider below 3 points:
Whether each cycle coupling has been fully completed, especially at the N terminus?
Whether the peptide needs to full reduction with EDT?
Whether have you dissolved it well before HPLC preparation?

hanieh ariani · #5 (**permalink**) 12-25-2008, 07:12 AM

Dear glbiochem
really thanks for your attention and help

zjuer · #6 (**permalink**) 12-31-2008, 03:02 PM

you can try to use new HPLC column,
or add some acetic acid or TFA to the sample assite dissovled.

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synthetic peptide purification , help

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