Multi-dimensional plug-and-play liquid chromatography-native ion mobility mass spectrometry method for the analysis of biotherapeutics
0301 basic medicine
03 medical and health sciences
01 natural sciences
0104 chemical sciences
DOI:
10.1016/j.ijms.2021.116725
Publication Date:
2021-10-09T12:03:17Z
AUTHORS (5)
ABSTRACT
Abstract A flexible inline size exclusion chromatography (SEC) and ion-exchange chromatography (IEX) method coupled to native mass spectrometry (nMS) method was developed for the analysis of protein aggregates and charge variants in a single run of biotherapeutics. This method can be applied to a broad spectrum of biologics such as human growth hormone and its soluble aggregates, the oligomer state distribution of liraglutide in solutions with different pH, and the native states of homodimers and heterodimers generated during a force stressed study of mixtures of two monoclonal antibodies. Tuning of both orbitrap and time of flight (TOF) mass spectrometers in native MS mode to accommodate this LC system is shown in this work for the analysis of biopharmaceutics. This SEC-IEX -UV-nMS system offers two-dimensional separation and two detection methods (ultraviolet absorbance and mass detection), providing an orthogonal characterization of complicated protein samples in a single method.
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