Interference‐free mass spectrometric detection of capillary isoelectric focused proteins, including charge variants of a model monoclonal antibody

摘要: CIEF represents an elegant technique especially for the separation of structural similar analytes, whereas MS is a state-of-the-art instrumentation identification and characterization biomolecules. The combination both techniques can be realized by hyphenating with CZE-ESI-MS applying mechanical valve. During CZE step, remaining ESI-interfering components electrolyte are separated from analytes prior to detection. In this work, multiple heart-cut approach presented expanding our previous single concept resulting in dramatical reduction analysis time. Moreover, different sample transfer loop volumes systematically compared discussed regard peak width efficiency. With major enhancement, model proteins (1.63-9.75 mg/L), covering wide pI range (5-10), charge variants deglycosylated antibody were analyzed on intact level. promising CIEF-CZE-MS setup expected applicable bioanalytical fields, e.g. fast information rich therapeutic antibodies.