Quantitative characterization of virus-like particles by asymmetrical flow field flow fractionation, electrospray differential mobility analysis, and transmission electron microscopy.
Here we characterize virus-like particles (VLPs) by three very distinct, orthogonal, and quantitative techniques: electrospray differential mobility analysis (ES-DMA), asymmetric flow field-flow fractionation with multi-angle light scattering detection (AFFFF-MALS) and transmission electron microscopy (TEM). VLPs are biomolecular particles assembled from viral proteins with applications ranging from synthetic vaccines to vectors for delivery of gene and drug therapies. VLPs may have polydispersed, multimodal size distributions, where the size distribution can be altered by subtle changes in the production process. These three techniques detect subtle size differences in VLPs derived from the non-enveloped murine polyomavirus (MPV) following: (i) functionalization of the surface of VLPs with an influenza viral peptide fragment; (ii) packaging of foreign protein internally within the VLPs; and (iii) packaging of genomic DNA internally within the VLPs. These results demonstrate that ES-DMA and AFFFF-MALS are able to quantitatively determine VLP size distributions with greater rapidity and statistical significance than TEM, providing useful technologies for product development and process analytics.
Pease
Leonard F
LF
National Institute of Standards and Technology (NIST), Gaithersburg, Maryland, USA.
Lipin
Daniel I
DI
Tsai
De-Hao
DH
Zachariah
Michael R
MR
Lua
Linda H L
LH
Tarlov
Michael J
MJ
Middelberg
Anton P J
AP
D000818:Animals; D001717:Birds; D002213:Capsid / Q000737:chemistry* / Q000648:ultrastructure; D004247:DNA / Q000235:genetics / Q000378:metabolism; D031521:Fractionation, Field Flow; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D009980:Influenza A virus; D007313:Insecta; D046529:Microscopy, Electron, Transmission; D010316:Particle Size; D011120:Polyomavirus; D011994:Recombinant Proteins; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011994:Recombinant Proteins; D014764:Viral Proteins; D004247:DNA