Rapid analytical and preparative separation of structural polypeptides of poliovirus by reverse-phase...

A reverse-phase high-performance liquid-chromatography (RP-HPLC) technique was developed for rapid (20 min) and effective separation of the structural polypeptides of poliovirus of strains of all three serological types. The method is useful for quantitative analysis as well as for isolation of polypeptides on a micropreparative scale for chemical, biochemical and immunological studies. All four virus polypeptides (VP1 to VP4) were obtained quantitatively in high purity by this one-step procedure. The solvents used were volatile and easily removed by evaporation, giving dry, amorphous polypeptides free of any additives. The separation mechanism of this RP-HPLC is based on the hydrophobicity of the protein surface. It is distinct from other separation methods such as molecular sieving and isoelectric focusing. Therefore, differences in primary and secondary structure of a polypeptide can be detected by RP-HPLC.