Solid Phase peptide synthesis (SPPS) ..

Solid Phase peptide synthesis (SPPS) is based on sequential addition of alpha-amino and side-chain protected amino acid residues to an insoluble polymeric support .

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Currently,“peptide synthesis” includes a large range of techniques and procedures that enable the preparation of materials ranging from small peptides to large proteins. The pioneering work of Bruce Merrifield, which introduced solid phase peptide synthesis (SPPS), dramatically changed the strategy of peptide synthesis and simplified the tedious and demanding steps of purification associated with solution phase synthesis. Moreover, Merrifield’s SPPS also permitted the development of automation and the extensive range of robotic instrumentation now available. After defining a synthesis strategy and programming the amino acid sequence of peptides, machines can automatically perform all the synthesis steps required to prepare multiple peptide samples. SPPS has now become the method of choice to produce peptides, although solution phase synthesis can still be useful for large-scale production of a given peptide.

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Biotin can be linked to the N-terminus of peptides or to the side chains of lysine (Lys) or glutamic acid (Glu) during Fmoc solid phase synthesis. Post synthesis biotinylation can be carried out using the side chain thiol of a cysteine (Cys) residue. This is particularly useful when a cysteine-containing peptide has been synthesised for conjugation to a carrier protein for antibody production.