Analysis of Bio-nanoparticles by Means of Nano ES in Combination with DMA and PDMA: Intact Viruses, Virus-Like-Particles and Vaccine Particles
For characterization of whole viruses, vaccine particles and virus-like-particles (VLPs) besides immunological and functional parameters usually methods as electron microscopy (EM), liquid phase separation or light scattering techniques are applied. The use of nano electrospraying (nano ES) to transfer such bio-nanoparticles (NP) from the liquid phase into the gas phase and ionization is a relative new development, i.e. to bring such kind of nano-objects as intact species into the gas-phase at atmospheric pressure. Now it is possible to generate ions with multiple charges as well as a single charge fixed on such spherical bio-NPs.
Here, we want to present two techniques which open up new avenues of analysis of intact viruses, VLPs and vaccine particles. Namely, a nano electrospray (nano ES) source with a charge manipulation (reduction) device is coupled to a separation device called differential mobility analyzer (DMA) followed by a detection system (either a condensation particle counter or Faraday cup). Two types of separation devices either a single DMA (gas-phase electrophoretic mobility macromolecular analyzer, GEMMA) or two in parallel used DMAs (PDMA) will be described and discussed. The size-separated bio-NPs are transferred in most cases then into the universal detector CPC (condensation particle counter). The other very interesting part of both instruments is the possibility to collect size-separated bio-NPs for subsequent further characterization as image generation by means of atomic force microscopy (AFM) or immunological evaluation by means of DotBlot via a specific antibody. Both devices will be compared and their development will be described. Both systems are allowing the handling of bio-NPs from 2.5 to several hundred nm and are measuring number concentrations. The characterization by means of nano ES GEMMA with sample collection after DMA separation and nano ES PDMA of intact viruses – human rhinovirus (HRV serotype 2; so-called “common cold” virus), of inactivated viruses – tick-borne encephalitis virus (TBEV) vaccines and of VLPs will be shown.
KeywordsBio-nanoparticles Viruses VLP Vaccines Size determination DMA GEMMA MacroIMS PDMA Particle collections AFM EM Immuno detection
The work regarding the PDMA was supported by grant of the Austrian Science Foundation (TRP29-N20 to W.W.S and G.A.). Furthermore we appreciate for making the following samples available – the hepatitis B virus capsid (A. Heck), TBEV vaccine (C. Tauer) and HRV2 (D. Blaas). The authors express their thanks for the AFM picture of the sample to G. Friedbacher and E. Eitenberger.
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