INVESTIGATIONS OF INTACT VIRUSES, VACCINES AND VIRUS-LIKE-PARTICLES BY MEANS OF NANO ES(I) COMBINED WITH A DIFFERENTIAL MOBILITY AND QqRTOF ANALYZER

摘要

For characterization of whole viruses, vaccines and virus-like-particles (VLPs) besides immunological and functional parameters usually methods as transmission electron microscopy (TEM), scanning electron microscopy (SEM), atomic force microscopy (AFM) in the tapping mode, analytical ultracentrifugation, size exclusion chromatography (SEC), asymmetric flow field-flow fractionation (AF4), multiangle light scattering (MALS) or dynamic light scattering (DLS) are used. Here, we want to present a workflow which is starting with SEC or dialysis followed by off-line nano electrospray (nES) in the cone-jet mode or classical nano ES ionization (nESI). The first device (nES) is integrated with a charge reduction chamber (Po-210) and coupled to a nano differential mobility analyzer (DMA; separation device working down to2.5 nm particle size) connected to condensation particle counter (CPC; allowing detection on the single particle level). This instrument is called gas-phase electrophoretic mobility macromolecular analyzer (GEMMA), a separation device allowing the handling of particles from 2.5 to several hundred nm and measuring number concentrations. The second device is a standard nano ESI source with differential pumped transfer system connected a 32 kDa quadrupole followed by a reflectron analyzer. Some bionanoparticle applications and the limits of both instruments will be presented. The characterization by means of nES GEMMA and nESI ion mobility QqRTOF mass spectrometry of intact viruses - human rhinovirus (HRV serotype 2; so-called "common cold" virus), of inactivated viruses - vaccines and of VLPs - human papilloma virus and hepatitis B virus shells will be shown.