TC-83-derived alphavirus vectors, particles and methods
AssigneeAlphaVax, Inc.
InventorsJon O. Rayner, Jonathan F. Smith, Bolyn Hubby, Elizabeth A. Reap
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Abstract
The present disclosure provides TC-83 VEE-derived replicons, alphaviral replicon particles and immunogenic compositions containing TC-83 alphaviral replicon particles which direct the expression of at least one antigen when introduced into a suitable host cell. The TC-83 VEE-derived ARPs described herein are improved in that they are subject to a lower vector-specific immune response than prior art ARPs.
Claims14
A method for preparing propagation-defective TC-83 derived alphaviral replicon particles (ARPs), said method comprising:
- (a) introducing an alphaviral replicon nucleic acid into a host cell, said replicon nucleic acid comprising at least a virus packaging signal and at least one heterologous coding or functional sequence expressible in said alphaviral replicon nucleic acid, said replicon nucleic acid lacking sequences encoding an alphavirus structural protein, wherein said host cell comprises at least one helper function encoding the TC-83 structural proteins comprising the TC-83 capsid protein and the TC-83 glycoproteins, to produce a modified host cell;
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- (b) culturing said modified host cell under conditions allowing expression of the at least one helper function, allowing replication of said TC-83-derived alphaviral replicon nucleic acid and packaging of said alphaviral replicon nucleic acid to form ARPs.
A method of preparing propagation-defective TC-83 derived alphavirus replicon particles comprising introducing an alphavirus replicon vector and one or more helper nucleic acid molecules into alphavirus-permissible cells via electroporation, wherein said alphavirus replicon vector comprises a nucleic acid lacking sequences encoding an alphavirus structural protein, wherein said one or more helper nucleic acid molecules encode the TC-83 structural proteins comprising the TC-83 capsid protein and the TC-83 glycoproteins, and wherein the alphavirus-permissive cells in a culture medium during electroporation are at a concentration of least 108 cells/ml medium and wherein the alphavirus RNA replicon vector is added to the cells prior to electroporation at a concentration of approximately 35 μg per ml.
Dependencies are inferred from the claim text.