Peptide Analysis by Rapid, Orthogonal Technologies with High Separation Selectivities and Sensitivities

Abstract

This article examines the current status of peptide analysis by orthogonal micro-/nano-separation strategies, with emphasis on the complementary use of high performance capillary liquid chromatography (μ -HPLC), capillary zonal electrophoresis (HPCZE), open tubular capillary electrochromatography (ot -CEC) and packed capillary electrochromatography (p -CEC). The ability to interface these techniques with mass spectroscopic (MS) procedures has enabled substantial progress to be made in the analysis of very small quantities of peptides, as well as proteins and other bio-macromolecules. As a consequence, the staged application of these high resolution techniques as part of the standardisation of biological products via robust, sensitive protocols is rapidly becoming a reality. Recent conceptual and theoretical advances have also allowed improved levels of prediction and optimisation of these procedures. Since significant differences in selectivity can be achieved with μ -HPLC, HPCZE and HPCEC respectively, collectively these sophisticated techniques provide unprecedented opportunities for the rapid, orthogonal and sensitive separation of complex mixtures of peptides and proteins. Several advantages of using these technologies in tandem are highlighted.