Freeze-Dried Composition of Inactivated Virus Envelope with Membrane Fusion Activity

Abstract

The objects of the present invention are to provide a freeze-dried composition of an inactivated virus envelop having membrane fusion activity which can be stored at higher temperatures without losing the ability to introduce foreign matters and to provide a method of introducing a foreign matter into a cell with high efficiency. The present invention provides a freeze-dried composition for introducing a foreign matter which comprises an inactivated virus envelope having membrane fusion activity, and at least one stabilizer selected from the group consisting of a protein hydrolysate, leucine, an L-arginine-acid and a polysaccharide, and a method of introducing a foreign matter using the freeze-dried composition containing an inactivated virus envelope.

1 . A freeze-dried composition comprising an inactivated virus envelope having membrane fusion activity, and at least one stabilizer selected from the group consisting of a protein hydrolysate, leucine, an L-arginine-acid and a polysaccharide. 
     
     
         2 . The freeze-dried composition according to  claim 1 , wherein the inactivated virus envelope is an inactivated virus envelope of a virus of Paramyxoviridae, Orthomyxoviridae, Herpesviridae, Hepadnaviridae or Flaviviridae. 
     
     
         3 . The freeze-dried composition according to  claim 1  or  2 , wherein the stabilizer is a protein hydrolysate and a polysaccharide. 
     
     
         4 . The freeze-dried composition according to any one of  claims 1  to  3 , which is prepared by a process comprising a step (1) of mixing a suspension of the inactivated virus envelope having membrane fusion activity and the stabilizer and a step (2) of freeze-drying the mixture obtained in the step (1). 
     
     
         5 . The freeze-dried composition according to  claim 4 , wherein the concentration of the inactivated virus envelope in the mixture obtained in the step (1) corresponds to an OD of from 0.1 to 7.0. 
     
     
         6 . The freeze-dried composition according to  claim 4  or  5 , wherein the concentration of the protein hydrolysate, leucine or the L-arginine-acid in the mixture obtained in the step (1) is from 0.1 to 2.5%. 
     
     
         7 . The freeze-dried composition according to any one of  claims 4  to  6 , wherein the concentration of the polysaccharide in the mixture obtained in the step (1) is from 0.05 to 0.5%. 
     
     
         8 . The freeze-dried composition according to any one of  claims 4  to  7 , wherein the salt concentration in the mixture obtained in the step (1) is at most 3 mM. 
     
     
         9 . A method of introducing a foreign matter into a cell or a living organism, which comprises: 
       (1) a step of rehydrating a freeze-dried composition containing an inactivated virus envelope having membrane fusion activity with a mixture comprising the foreign matter and water, and 
       (2) a step of bringing the composition obtained in the step (1) into contact with the cell or the living organism. 
     
     
         10 . The method of introducing a foreign matter according to  claim 9 , wherein the freeze-dried composition is the freeze-dried composition as defined in any one of  claims 1  to  8 . 
     
     
         11 . A cell fusion method comprising: 
       (a) step of rehydrating a freeze-dried composition containing an inactivated virus envelope having membrane fusion activity; and 
       (b) step of mixing the composition obtained in the step (a) with a suspension of cells to be fused. 
     
     
         12 . The cell fusion method according to  claim 11 , wherein the freeze-dried composition is the freeze-dried composition as defined in any one of  claims 1  to  8 .