DANE, CAMERON and BRIGGS (1970) identified 42-nm "Dane" particles in the serum of individuals infected with hepatitis B virus (HBV), and they have been accepted as representing HBV. Dane particles share hepatitis B surface antigen (HBAg) with 20-nm spherical and tubular forms, but their cores, hepatitis B core antigen (HBAg), have a distinct antigenicity (Almeida, Rubenstein and Stott, 1971). We have developed a method for the isolation of Dane particles from the plasma of asymptomatic carriers of HBAg on a large scale, and demonstrated a double-stranded DNA molecule extruding directly from their cores (Takahashi, T. , 1976). In the course of this study, we realised that the cores of Dane particles shedding a DNA strand were surprisingly few; more than 90% of Dane particles were thought to be "empty" because they did not seem to contain any DNA strand. The separation of the Dane particles containing DNA from such defective ones, to create a homogenous population of complete HBV, would be a prerequisite for the study and understanding of their physicochemical properties, such as the DNA structure.

Birnie, Rickwood and Hell (1973) observed that the density of DNA decreased remarkably in the presence of metrizamide due to its hydration. They noted that DNA had a density of 1.1 g/cm in metrizamide solution, as against 1.7 g/cm in CsCl solution. Taking advantage of this phenomenon, we tried to separate the cores of Dane particles containing DNA from those without DNA.


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