Vm. Okun et al., Analysis of common cold virus (human rhinovirus serotype 2) by capillary zone electrophoresis: The problem of peak identification, ANALYT CHEM, 71(10), 1999, pp. 2028-2032
Different preparations of human rhinovirus serotype 2 (HRV2), a common cold
virus, were analyzed by capillary zone electrophoresis (CZE) in untreated
fused-silica capillaries using berate buffer (100 mmol/L, pH 8.3) and sodiu
m dodecyl sulfate (10 mmol/L) as additive to prevent wall adsorption. The e
lectropherograms showed one major peak at 205- and 254-mm detection wavelen
gths. The identity of the peak as originating from native virus was confirm
ed by several indirect methods. Heating to 56 degrees C is known to lead to
release of the genomic RNA from the viral capsid; this treatment resulted
in the disappearance of the major peak and the emergence of a new predomina
nt peak that was identified as RNA by enzymatic digestion. As expected, RNa
se treatment of the unheated sample remained without effect as the viral ge
nome is inaccessible in the native viral shell. A monoclonal, virus-aggrega
ting antibody was used for immunodepletion of native vints; again, the majo
r peak disappeared upon removal of viral aggregates by centrifugation prior
to CZE analysis. In combination, these results allowed for the unambiguous
identification of the main peak as native HRV2 and of the minor peaks as c
ontaminants present in various amounts in the different viral preparations.
It is demonstrated that CZE allows for an extremely easy and rapid assessm
ent of conformational state and purity of virions in a given viral preparat
ion.