Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4548

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Purification of Peptides from Solid-Phase Peptide Synthesis with RP-HPLC

Reinhard I. Boysen and Milton T.W. Hearn

This protocol was adapted from "Reversed-Phase High-Performance Liquid Chromatography," Chapter 5 in Proteins and Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.


INTRODUCTION

Purification of peptides derived from solid-phase peptide synthesis (SPPS) requires the removal of deletion peptides resulting from incomplete coupling/deprotection steps, from racemization or side-chain rearrangement, and from various chemical substances introduced during the deprotection or cleavage stages of an SPPS procedure. This protocol details the purification and analysis of many synthetic peptides of 2-65 amino acid residues. These peptides contain a number of ionizable or polar side chains, but do not contain secondary structural elements (such as ß-sheets) that favor supramolecular assembly.


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