Alphavirus particles and methods for preparation

US7078218B2 · kind B2 · utility

Assignee

• ALPHAVAX, INC. · US

Inventors

• Jonathan F. Smith · Cary, US
• Kurt Iver Kamrud · Apex, US
• Sergey A. Dryga · Chapel Hill, US
• Harold Alterson · Cary, US
• Jon O. Rayner · Apex, US
• Kim Butler · Cary, US
• Maureen Maughan · Durham, US

Key dates

• Filing date: Dec 12, 2003
• Grant date: Jul 18, 2006
• Expiry date: Dec 12, 2023

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2770/36162
• WIPO field: Pharmaceuticals
• WIPO sector: Chemistry

Abstract

Provided herein are methods for producing alphavirus replicon particles in high yield; replicon RNAs are electroporated into permissive cells, where the cells are at a relatively high density, together with at least one helper nucleic acid providing the necessary functions for packaging. After a growth period in appropriate medium, alphavirus replicon particles are harvested from the surfaces of the cells in which they were produced using a salt wash in which the salt concentration is from about 0.2 to about 5 M sodium chloride, calcium chloride, magnesium chloride, potassium chloride, ammonium acetate, ammonium bicarbonate, among others. After dilution, if necessary, the particles can be purified by a suitable chromatographic technique.