Analysis of common cold virus (human rhinovirus serotype 2) by capillary zone electrophoresis: The problem of peak identification

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.