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AssigneeAlphaVax, Inc.
InventorsJonathan F. Smith, Kurt Kamrud, Sergey Dryga, Harold Alterson, Jon Rayner, Kim Butler, Maureen F. Maughan
Estimating the term…
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.
A method for preparing alphaviral replicon particles (ARPS), said method comprising the steps of:
A method of preparing alphavirus replicon particles comprising introducing an alphavirus replicon vector and one or more helper nucleic acid molecules into alphavirus-permissive cells via electroporation, wherein concentration of the alphavirus permissive cells in culture medium during electroporation is from 5×107 to 5×108 cells/mL and wherein the concentration of the alphavirus RNA replicon vector added to the cells prior to electroporation is approximately 35 μg per mL.
A method for preparing alphaviral replicon particles (ARPS), said method comprising the steps of:
Dependencies are inferred from the claim text.