During attempts at purification of horse-sickness virus for electron-microscopy from infected mouse brain extracts, it was found that a combination of polyethylene glycol precipitation of the virus, ultracentrifugation and zone electrophoresis gave promising results. The final purified material could not be regarded as pure on account of the presence of considerable normal brain components, but it contained particles not present in normal brain treated identically, which could be recognized as virus particles. The virus particles appear to have diameters of 70–80 mμ and are estimated to have 92 rod-shaped subunits radiating from a spherical body.The authors wish to express their gratitude to Prof. A. Kipps and R. A. Alexander for their continued interest in this work.The electron micrographs were taken by Mr L. G. Fowle of the Department of Physics, University of Cape Town.This investigation was supported in part by a Public Health Service research grant AI 04044–02 from the National Institutes of Health, Bethesda, U.S.A.
Zone electrophoresis in a sugar concentration gradient is proposed as a complementary method for the biological classification of the enteroviruses. Under standardized experimental conditions it was found that the majority of the Coxsackie A viruses and the polioviruses have a common, low electrophoretic mobility. sackie A viruses and the polioviruses have a common, low electrophoretic mobility. The impression was gained that the ECHO viruses show the highest electrophoretic mobility, though in some cases there is overlapping with the Coxsackie Bs. The mobility of both of these groups is considerably higher than that of the other two.Zone electrophoresis appears to be an excellent method for the purification of the enteroviruses prior to the production of specific precipitating antibodies.
It was shown in this work that the electrophoretic mobility of the MEF1 strain of poliovirus was increased approximately 25% after interrupted inactivation with formaldehyde. This is in accordance with past observations on the behaviour of formaldehyde-treated proteins. The residual live virus had the same mobility as the ‘killed’ virus, thus indicating that the surfaces of all the virus particles have been affected to the same degree by the inactivating agent.Gel precipitin tests performed on the different samples obtained by zone electro-phoresis showed, in addition to the main antigen, the presence of a small amount of a second component of slightly higher mobility, and, as it occurs in the main fraction and the sample thereafter it probably has a slightly higher mobility than the main component.The antigenic properties of the different fractions as shown by the production of antibodies in guinea-pigs correspond with the presence of precipitating antigen in the samples in the region of highest antigen concentration. The faint antigenicities of the fractions obtained from the regions higher up in the zone electrophoresis column are due to small amounts of virus antigen similar to that found in the column on electrophoresis of untreated MEF1 virus, bearing in mind that the inactivated virus was concentrated approximately 2000 times for zone electrophoresis.
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