Large-scale isolation of Dane particles from plasma containing hepatitis B antigen and deomnstration of circular double-stranded DNA molecule extruding directly from their cores.
A method was developed to isolate, on a large scale, Dane particles from five liters of pooled plasma of asymptomatic carriers of hepatitis B antigen. It involved three successive ultracentrifugation procedures; the final preparation was more than 98% pure in Dane particles. Purified Dane particles had the density of 1.23 to 1.24 g/cm3. The concentration of nucleic acids in the preparation containing purified Dane particles was too low to be detected either by chemical or by spectrophotometric method. However, a circular double-stranded DNA molecule extruding directly from the core of Dane particles was clearly demonstrated by elelctron microscopic observations. The cores of Dane particles (hepatitis B core antigen; HBCAg) were prepared by treating the Dane particle preparation with mercaptoethanol and Nonidet P-40. They were sufficient both in purity and amounts to allow the determination of antibody to HBCAg by an immune adherence hamagglutination method.
D000818:Animals; D002353:Carrier State / Q000276:immunology*; D002501:Centrifugation, Zonal; D004247:DNA / Q000032:analysis*; D006168:Guinea Pigs; D006511:Hepatitis B Antigens / Q000302:isolation & purification*; D006512:Hepatitis B Core Antigens; D006801:Humans; D007114:Immunization; D011817:Rabbits; D014461:Ultracentrifugation
D006511:Hepatitis B Antigens; D006512:Hepatitis B Core Antigens; D004247:DNA