A simple method for isolation of cell-associated viral particles from cell culture

• Published in: Journal of virological methods 2017-11, Vol.249, p.194-196
• Main Authors: Laposova, Katarina, Oveckova, Ingrid, Tomaskova, Jana
• Format: Article
• Language: English
• Subjects: A549 Cells; Animals; Antibodies, Viral; Buffers; Cell Culture Techniques; Cell-associated virus; Cell-free extract; Cercopithecus aethiops; Culture Media - chemistry; Freezing; HeLa Cells; Humans; Infection; LCMV; Lymphocytic Choriomeningitis - diagnosis; Lymphocytic Choriomeningitis - virology; Lymphocytic choriomeningitis virus - growth & development; Lymphocytic choriomeningitis virus - isolation & purification; Mice; Nucleoproteins - isolation & purification; Vero Cells; Viral particles; Virion - isolation & purification; Virology - methods; Virus isolation
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2017.09.014

•Compared hypotonic buffer, water, freeze-thaw and sonication to obtain cell-associated virus.•Isolation method using deionized water lysis only is the most effective.•Freeze-thaw cycles and sonication do not improve cell-associated virus isolation. A common method for cell-associated virus isolation involves disruption of infected cells by a combination of hypotonic burst, freeze-thaw cycles (F-T) and sonication. This protocol was also originally used for the preparation of cell-free extract containing the MX strain of lymphocytic choriomeningitis virus (LCMV), which is preferentially propagated by cell-to-cell contact and does not release distinct virions into the medium. In the present study, we compared different approaches to virus isolation. Based on virus yield, we show that deionized water lysis is the fastest and most effective method for releasing LCMV MX infectious viral particles from persistently infected cells. Moreover, we demonstrate that freeze-thaw cycles and sonication do not improve virus isolation. This simple protocol could be used for isolation of other viruses, the life cycle of which is strictly cell-associated and therefore are difficult to release in large amounts from host cells.