I produced recombinant LL-37 and I want to try antibacterial activity of this product. However I cant measure antibacterial of recombinant LL-37, but purchased synthetic LL-37 have expected activity (according to old articles). I analyze my sample of recombinant LL-37 on MS MALDI-TOF, everething is OK (I see peak about 4 600 Da = GLL-37). Has anyone meet with this problem? Maybe works: Lyophilized peptide LL-37 (in solution 25 mM TRIS, 25 mM NaCl, 1 mM DTT and pH = 8) and lyof. powder dissolved in organic compounds (acetonitrile, TFA), after that, again lyophilized and after that, use for measure antimicrobial activity?
Dear Antonin,
might be a tertiary structure issue due to denaturation. Do you have an opportunity to compare your material with the commercial sample by circular dichroism?
Do you have any clues about the additives of the commercial product?
LL-37 a peptide and form a only alfa helix, but if LL-37 doesnt form alfa-helix = no activity (i know helix is important for antimicrobial activity for this peptide).
I also agree with the denaturation hypothesis: Could you test 2-propanol instead of acetonitrile?
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