oligonucleotide-polypetide conjugation

WS via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Fri Apr 20 05:10:40 EST 2012


Hi Alexandra,

DMSO should work fine. Actually, anything that does not react with the
formyl group and with the hydrazine, should work (basically, you might
avoid eg acetone and possily alkohols, also check for possible
denaturants like MEK)

You might consider to keep the Cys definitely reduced by adding eg
some TCEP.

If this is the only Cys in your peptide, why not simply use a
maleimido-labeled oligo?

Other suitable nucleotide labels available are triple bonds or
nitriles, which are easily clicked to azides. IIRC, maleimido-spacers
are available with a terminal azide.

You might check Pierce for linkers an have a look at metabion.com for
possible oligo modifications.

Have fun(ds) and good luck!

Wo

On 19 Apr., 23:20, alexandra balliou <alexandra.ball... from gmail.com>
wrote:
> First of all, I would like to congratulate for this so helpful forum.
> I am a Ph.D student at the department of organic chemistry and
> biochemistry. I am starting now my studies and a part of my project is
> to find a method to conjugate a 14 mer oligonucleotide with a 42
> residues polypeptide helix-loop-helix conformation. I am thinking of
> conjugating them by a hydrazone bound, where a maleimide-hydrazine is
> incorporated in the cysteine residue at the loop region and a
> formylbenzoate group modifies the oligonucleotide. The reaction is SN2
> so I was thinking of using DMSO as a solvent for the conjugation in
> combination with tetra t-butyl ammonium counterions. Do you think that
> the method  will work? I am looking forward to hearing from you. Thank
> you in advance!



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