Analyzing complex mixtures of polypeptides, useful e.g. for proteome analysis, by fragmentation, immobilization, selective release, labeling and characterization

摘要

Analyzing complex mixtures of biopolymers (I) that contain peptide bonds by fragmentation, immobilization, selective release of bound fragments, labeling, separation of labeled fragments according to physicochemical properties and detecting the label. Analyzing complex mixtures of biopolymers (I) that contain peptide bonds comprises: (a) cleaving (I) to fragments (F), using one or more chemical reagents and/or enzymes; (b) coupling at least some (F) and/or (I) to an insoluble carrier by covalent attachment to a linker, already bound to an anchor group on the carrier; (c) washing to remove unbound (F) and/or (I); (d) selective decoupling of bound (F) and/or (I) by cleaving either the bond between linker and (F) or (I), or a bond within (F) and/or (I); (e) covalent attachment of a marker (M) to selected (or all) uncoupled or decoupled (F), where (M) makes possible detection of (F) on the basis of chemical or physical properties that are not present, or if present are distinguishably different, in unmarked (F) and/or (I); and (f) separating labeled (F) on the basis of physicochemical properties and from their labeling. An Independent claim is also included for a kit for performing the method.