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Purification of Herpes Simplex Virus

Department of Virology Institute of Medical Microbiology University of Göteborg, Göteborg, Sweden

Herpes simplex virus was highly purified from host cell material with little loss of infectivity by the combination of isopycnic centrifuging in a three layered silica gradient and rate zonal centrifuging in dextran T 70 gradient. By layering three solutions of colloidal silica (24, 18 and 12%, w/v) on top of each other in the centrifuge tube prior to ultracentrifuging, a linear type of gradient was obtained. Virus particles and nucleocapsids seemed to be distinctly separated in this three-layered silica gradient. The overall reduction of host proteins and sphingolipids was 1250 to 2000 times and 350 to 600 times, respectively. The reduction of virus specified non-particulate proteins was approx. 500 times. The recovery of infectivity was 55 to 70%.

Received 16 July 1973; accepted 29 October 1973.

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