Virus analysis using electromigration techniques.
We discuss the progress during the last 4 years in the analysis of viruses by electrophoresis in capillaries and microfluidic devices. The paper is the continuation of a review published in this journal in 2005 [Kremser, L., Blaas, D., Kenndler, E., Electrophoresis 2004, 25, 2282-2291]. Eighteen papers on the topic have appeared since; the majority deals with zone electrophoresis and three reports are on IEF. These methods have been applied to human rhinoviruses, poliovirus Semliki Forest virus, norovirus-like particles, and the two bacteriophages MS2 and T5. A main finding was that addition of detergents and salts to the BGEs are essential for the robustness of the CE analysis. Analyte detection was usually via UV absorbance but there are some examples where the viruses were rendered fluorescent via modification of the capsid proteins with reactive dyes and/or by non-covalent attachment of intercalating fluorescent compounds to the nucleic acids making up the viral genome. Interestingly, some viruses are permeable to small molecular mass components; this allows fluorescent dyes to diffuse into the intact virus where they attach to the nucleic acid. Release of a viral genome upon heating was also monitored by using similar methodologies. Interactions of viruses and subviral particles with antibodies, receptors, and receptor-decorated liposomes were investigated with CE methods, all by using a non-equilibrium approach (i.e. co-incubation of the components prior to CE separation). Viruses are multivalent (i.e. possess many identical surface-exposed patches) and most of them are composed of defined numbers of identical subunits. The high resolution of CE has been most remarkably demonstrated by the separation of stoichiometric complexes between virus and a distinct number of soluble recombinant receptors and revealed their concentration-dependent distribution.
Kremser
Leopold
L
Innsbruck Medical University, Biocenter, Division of Clinical Biochemistry, Innsbruck, Austria. Leopold.Kremser@i-med.ac.at
Blaas
Dieter
D
Kenndler
Ernst
E
D004279:DNA, Viral / Q000032:analysis; D019075:Electrophoresis, Capillary / Q000379:methods*; D046209:Electrophoresis, Microchip / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D012367:RNA, Viral / Q000032:analysis; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D014764:Viral Proteins / Q000032:analysis; D014780:Viruses / Q000737:chemistry*
D004279:DNA, Viral; D012367:RNA, Viral; D014764:Viral Proteins