Abstract
Because immunoassay responds to epitopes, and many molecules share the same peptide epitope, it is very difficult to obtain an accurate understanding of peptides, their creation and hydrolysis, in biological systems. Separate-and-detect approaches have merit in that the many active peptides and inactive fragments of a particular system can be separately determined. This review discusses the separation, by chromatography and capillary electrophoresis, and detection, by absorbance, fluorescence, electrochemistry, and immunoassay techniques. When separation pre-concentration is accompanied by laser-induced fluorescence or biuret-based electrochemical detection, nM-pM detection limits are obtained.