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CUSTOM PEPTIDE SYNTHESIS - Technical background
Proteogenix uses manual and automatic peptide synthesis technique. Manual technique is more flexible than automated synthesizer. We can therefore make much longer peptides and special modified peptides more easily.
Proteogenix uses solid phase peptide synthesis (SPPS) which consists of assembling amino acids from the C-terminal to the N-terminal. The a-carboxyl group is attached via an acid labile linker to a solid support, the “resin”.
The amino terminal end of the amino acid is protected by a base-labile Fmoc protecting group while the side chains are protected by acid-labile groups such as tertiary-butyl(tBu).
After the first aa is loaded onto the resin, the Fmoc group is removed using piperidine (Deprotection). A kaiser test is then performed to confirm that all of the Fmoc protecting groups are removed. The next Fmoc amino acid is then attached to the growing peptide by activation of its carboxyl group (Coupling). A kaiser test is then performed to confirm that complete coupling has occurred on all the free amines on the resin.
Synthesis then proceeds through a cycle of 1) deprotection of Fmoc amino terminus groups and 2) coupling of the next amino acid until the peptide is completely synthesized.
The completely synthesized peptide is then cleaved from the resin and side chain protection groups are removed using TFA (Cleavage).