ABSTRACT
The N-formyl group is the most commonly used protecting group used for tryptophan (Trp) during solid-phase peptide synthesis. This chapter describes the successful separation, in good yield, of the two bovine parathyroid growth hormone (bPTH) analogs following application of the shallow gradient approach. Two forms of a 28-residue bPTH were prepared: Ac-bPTH (7–34)-amide (P2) and Ac(N-For-Trp)-bPTH (7 to 34)-amide (P1). In addition, the utility of multiwavelength diode-array detection, coupled with gradient elution fraction analysis, during preparative peptide purification is highlighted. The chapter describes the successful preparative purification of ca. 20 mg of a crude peptide mixture of two closely-related 28-residue peptide analogs on an analytical column and with analytical instrumentation. Reversed-phase chromatography (RPC), with its powerful peptide resolving capabilities, was the logical method of choice for attempting to separate the peptides quickly and with satisfactory yields of purified peptides.
