Profiling of neuropeptides using gradient reversed-phase high-performance liquid chromatography with novel detection methodologies.
作者： Genevieve H. Fridland , Dominic M. Desiderio
DOI: 10.1016/S0378-4347(00)80686-6
关键词:
摘要: Biological tissues and fluids are subjected to gradient reversed-phase high-performance liquid chromatography (RP-HPLC) separation the neuropeptide profile of collected fractions is obtained by radioreceptor assay (RRA) using a broad-based competing ligand. Radioimmunoassay (RIA) also used detect specific neuropeptides in HPLC-purified fractions. Further confirmation identity peptides present tissue mass spectrometry (MS) fast atom bombardment (FAB) mode. FAB-MS produces protonated molecular ion peptide allows direct measurement underivatized at nanogram level, with increased specificity. FAB-MS-MS identifies unique amino acid sequence-determining spectrum offers maximum This analytical methodology applied study basis several disease states monitoring peptidergic pathways individual their metabolic relationships. Molecular mechanisms involved pain, stress, tumor formation, neurological studies studied.
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