RE: Arg-Pbf rescue

James Farmar (jfarmar@nybc.org)
Fri, 30 Jul 1999 13:59:29 -0400

Jane,
Two suggestions:
Are you sure that the Pbf has stayed with the Arg residue and not added
somewhere else in the peptide?
Have you tried adding the amino acid Arg to the cleave cocktail as an
additional scavenger?

Dr. Jim Farmar
Kimball Research Institute/NYBC

Hello all,

I have been cursed with a peptide which has some residual Pbf protecting
group left on an Arg residue (it is a 21-mer). The peptide synthesizes
beautifully, one peak on HPLC and usually one peak on MALDI-TOF but the
removal of the Pbf is somewhat hit and miss. I have never encountered a
problem with its removal before so I have been surprised to see it on this
peptide. I make a bunch of this stuff and the cleavage either goes to
completion or it gives me this portion of still Pbf protected peptide even
when batches are cleaved side by side.

As it happens, the Pbf protected bits elute at roughly the same % as the
clean stuff so I had hoped to try a rescue measure and re-cleave the already
cleaved peptide. I looked through the ABRF archive and could not find any
instance of this being attempted before so here I am, asking for your
assistance. I have had excellent results removing residual Trt from cleaved
peptides by stirring them in 99% TFA and 1% TIS, anyone have any experience
with Pbf removal in the same general fashion? I was thinking of using 95%
TFA + 2.5% EDT + 2.5% TIS.

Your input would be much appreciated!

Thanks in advance,

Jane

Jane A. Nagel MSc