The invention provides methods of synthesizing peptides, involving the steps of providing a composition including
a peptide fragment, wherein the
peptide fragment has at least one
amino acid residue and includes a base-sensitive, N-terminal
protecting group; removing the base-sensitive, N-terminal
protecting group from the
peptide fragment using a deprotection
reagent that includes a base, whereby an N-terminal functionality on the
peptide fragment is deprotected; removing the base from the composition to provide a residual base content of more than 100 ppm; causing a reactive
peptide fragment having a reactive C-terminus and a base-sensitive N-terminal
protecting group to react with the deprotected N-terminal functionality of the
peptide fragment under conditions such that the reactive
peptide fragment is added to the peptide fragment; and optionally repeating the deprotection and
coupling steps until a desired peptide is obtained. Also provided are methods of synthesizing peptides, wherein base is removed from the composition to a point where the composition would provide a positive
chloranil test. Also provided are methods of synthesizing peptides, wherein
coupling is performed in basic reaction mixtures.