The effect of anionic surfactant on poliovirus particles during capillary electrophoresis.
Because of its essential role in SDS-PAGE, sodium dodecylsulphate (SDS) is generally associated with protein denaturation. However, for SDS-PAGE, proteins are linearized in the presence of SDS, following the exposure to high temperatures and reducing agents. In comparison, the conditions employed during a capillary electrophoretic (CE) separation involve only a limited exposure to SDS, at much lower temperatures. As the outer surface of the non-enveloped viruses consists of proteins, virus interaction with SDS can be judged from the perspective of SDS-protein interaction. Several studies have indicated that proteins have a different susceptibility to SDS, depending on their secondary structure and number of subunits. Therefore it is not straightforward to estimate what should be expected when intact polioviruses and subviral particles obtained by thermal conversion of the poliovirions, are exposed to SDS during CE separation. In this study it is shown that, during CE separations, SDS has no effect on the integrity of the poliovirion, but the presence of SDS in the separation system influences the poliovirus peak height and shape. The implication of SDS in the CE separation of poliovirus is discussed in detail. On the contrary, the proteinaceous subviral particles, such as the empty capsids, are less stable in the presence of SDS during the CE separation, and aggregates between the individual poliovirus capsid proteins and SDS are formed. Finally, we have proposed an alternative separation approach, involving an SDS gradient, for an improved separation of the subviral particles.
Vrije Universiteit Brussel-VUB, Center for Pharmaceutical Research-CePhaR, Department of Analytical Chemistry and Pharmaceutical Technology, Laarbeeklaan 103, B-1090, Brussels, Belgium.
Iuliana
I
Oita
Hadewych
H
Halewyck
Bert
B
Thys
Bart
B
Rombaut
Yvan Vander
YV
Heyden
D000838:Anions / Q000737:chemistry; D002213:Capsid / Q000737:chemistry*; D036022:Capsid Proteins / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D017955:Poliovirus / Q000737:chemistry*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry*; D014764:Viral Proteins / Q000737:chemistry*
D000838:Anions; D036022:Capsid Proteins; D013501:Surface-Active Agents; D014764:Viral Proteins; D012967:Sodium Dodecyl Sulfate