Method for separating virus-like particles from a cell suspension

A method for separating virus-like particles from a cell suspension of host cells containing the virus-like particles, wherein the virus-like particles have at least one envelope protein embedded in a lipid double membrane, the protein comprising at least a portion corresponding to a small envelope protein of a virus of the family Hepadnaviridae, wherein

• a) the host cells are disrupted in order to obtain a first suspension containing the virus-like particles,
• b) a supernatant containing the virus-like particles is separated from the first suspension,
• c) an adsorbent is added to the supernatant separated from the first suspension such that the virus-like particles are adsorbed by the adsorbent,
• d) the adsorbent with the adsorbed virus-like particles is separated off,
• e) the virus-like particles are desorbed from the separated-off adsorbent, so that they pass into a supernatant of a second suspension formed thereby, and
• f) the supernatant containing the virus-like particles is separated from the second suspension,
• wherein subsequently the virus-like particles are separated from the separated-off supernatant and purified,
• wherein in step e) for desorbing the virus-like particles a desorption buffer is added which contains deoxycholic acid and/or at least one soluble salt of deoxycholic acid and
• wherein after step f):
• g) a soluble calcium salt is added to the separated-off supernatant, forming a precipitate containing a calcium salt of deoxycholic acid and the virus-like particles,
• h) the precipitate formed is separated off and transferred to a third suspension and
• i) the virus-like particles are separated from the third suspension and purified.