SELECTIVE PURIFICATION OF SYNTHETIC PROTEINS BY THE USE OF FMOC-BASEDAND BIOTIN-BASED REVERSIBLE CHROMATOGRAPHIC PROBES

Purification of proteins synthesised by the solid-phase stepwise appro ach, using conventional chromatographic tactics is often difficult and results in poor yields. Purification through selective labelling of t he target sequence with chromatographic probes when accompanied by an efficient capping protocol is an effective way to drastically improve both the quality and yields of these large, chemically synthesised pol ypeptides. Basically, the chromatographic probe consists of two parts, an 'affinity' group that significantly changes the properties of the peptide possessing it and a reversible linker through which the probe is attached to the peptide. Furthermore, the latter is stable under th e conditions used to deprotect and cleave the peptide from the resin s upport. After separation of labelled sequences from terminated peptide s using a suitable 'affinity' chromatographic method the label is sele ctively removed thus yielding the purified polypeptide. Several 'chrom atographic probes' have been proposed throughout the years. Those base d on the FMOC molecule and on the biotin/avidin interaction seem to be of more general applicability and are discussed here. (C) 1997 Elsevi er Science B.V.