Purification of adenovirus hexon by high performance liquid chromatography.
Hexon is the major structural protein of adenovirus, and has significance in studies of virus structure and function, vaccine development, and immunodiagnosis. We describe a simple, single-step, anion-exchange high performance liquid chromatography (HPLC) method for the high yield purification of hexon. Purity of the isolated hexon was assessed by SDS-PAGE and HPLC methods. The isolated hexon was immunologically reactive with anti-hexon monoclonal antibody in a dot-blot assay. It also retained immunogenicity, as polyclonal antisera from rabbits immunized with hexon showed the desired antigen specificity. The enhanced speed of this purification method allows for the efficient isolation of hexon from various serotypes, and thus may facilitate comparative studies of hexon immunobiology.
Siegel
S A
SA
Immunodiagnostics Department, Becton Dickinson and Company, Corporate Research Center, Research Triangle Park, North Carolina 27709.
Hutchins
J E
JE
Witt
D J
DJ
D000256:Adenoviridae / Q000032:analysis*; D002213:Capsid / Q000276:immunology / Q000302:isolation & purification*; D036022:Capsid Proteins; D002851:Chromatography, High Pressure Liquid; D006801:Humans
D036022:Capsid Proteins; C018315:hexon capsid protein, Adenovirus