SPPS (Solid-Phase Peptide Synthesis) is a fairly simple and reliable method
for the synthesis of peptides and smaller proteins. These are very important molecules
in nature as they perform many vital functions. The Fmoc group is most commonly used
as a temporary protection at the N-terminus of a modified amino acid during SPPS.
However, to control the synthesis and to build only one main polypeptide chain, all
other functional groups that could react during synthesis also need to be protected. We
prepared 2 peptides - one smaller by manual synthesis on a column (a.k.a. batchwise
method), as well as with continuous flow and without permanent protecting groups.
After successful preparation of the smaller peptide (12 amino acids) we attempted the
synthesis of a slightly longer peptide (26 amino acids). Following the established
protocol, the first modified amino acid was first coupled to the solid support, washed
with DMF, the Fmoc group was removed with piperidine in DMF, and washed again.
These cycles were repeated until all the required amino acids were included into the
chain. The peptide was constructed from the C- to N-terminus. The peptides were then
purified on a reverse-phase chromatographic column and analysed by HPLC and
HRMS.
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