Method for Producing Viral Vaccines

Abstract

The present invention provides a method for the manufacture of a preparation comprising virus antigens comprising a) inoculation of cells with infectious virus in a fluid, b) propagation of said virus in said cells, c) collecting said propagated virus, d) inactivating said collected virus, and e) treating said inactivated virus with a detergent, resulting in a preparation comprising viral antigens.

1 - 22 . (canceled) 
     
     
         23 . A method of increasing the resistance to a viral infection in a subject comprising administering a preparation comprising viral antigens, the preparation obtained by
 a method comprising   a) inoculating cells with infectious virus in a fluid,   b) propagating virus in said cells,   c) collecting said virus propagated in said cells,   d) completely inactivating said virus collected from said cells, and   e) purifying the inactivated virus, wherein the purification comprises ultracentrifugation or use of a sucrose gradient,   f) contacting the purified virus with a nuclease, and   g) treating said virus with detergent under conditions effective to yield a split virus,   resulting in a preparation comprising viral antigens, wherein the collected virus of step (c) is not purified prior to the inactivating step.   
     
     
         24 . The method of  claim 23 , wherein the step of collecting said propagated virus comprises separating the virus from said cells and/or cell debris of said cells after infection. 
     
     
         25 . The method of  claim 23 , wherein said inactivating is performed by addition of formaldehyde. 
     
     
         26 . The method of  claim 23 , wherein said inactivating is performed by UV irradiation. 
     
     
         27 . The method of  claim 23 , wherein said virus propagated in said cells is released into said fluid. 
     
     
         28 . The method of  claim 23 , wherein step (c) or (d) further comprises treating the virus with a nuclease. 
     
     
         29 . The method of  claim 28 , wherein said nuclease is Benzonase®. 
     
     
         30 . The method of  claim 23 , wherein said cells are in form of a cell culture during said virus propagation. 
     
     
         31 . The method of  claim 23 , wherein said cells are mammalian or avian cells. 
     
     
         32 . The method of  claim 23 , wherein said cells are epithelial cells. 
     
     
         33 . The method of  claim 31 , wherein said cells are Vero cells. 
     
     
         34 . The method of  claim 23 , wherein said virus is an enveloped virus. 
     
     
         35 . The method of  claim 34 , wherein said virus is an orthomyxo virus. 
     
     
         36 . The method of  claim 35 , wherein said virus is an influenza virus. 
     
     
         37 . The method of  claim 23 , wherein the concentration of non-viral protein during said inactivation is below 350 μg/ml. 
     
     
         38 . The method of  claim 23 , wherein said manufacture is on industrial scale amounts. 
     
     
         39 . The method of  claim 37 , wherein said inactivation is performed on at least 1 L virus containing fluid. 
     
     
         40 . A method of increasing the resistance to a viral infection in a subject comprising administering a preparation comprising viral antigens obtained by a method comprising
 a) obtaining a clarified supernatant comprising infectious virus,   b) inactivating said virus from step (a),   c) purifying the virus, wherein the purification comprises ultracentrifugation or use of a sucrose gradient,   d) contacting the inactivated, purified virus with a nuclease,   e) treating said virus with detergent under conditions effective to yield a split virus, and   f) further purifying said virus resulting in a preparation comprising viral antigens,   
       wherein the clarified virus of step (a) is not purified prior to the inactivating step. 
     
     
         41 . The method of  claim 40 , wherein said clarified supernatant is obtained from a cell culture. 
     
     
         42 . The method of  claim 41 , further comprising the step of stabilizing said viral antigens. 
     
     
         43 . The method of  claim 42 , wherein said viral antigens are stabilized by addition of an effective amount of Tween 80® (polysorbate 80). 
     
     
         44 . The method of  claim 43 , wherein Tween 80® is in an amount of about 0.125%. 
     
     
         45 . A vaccine comprising a preparation of viral antigens, the preparation obtained by a method comprising
 a) inoculating cells with infectious virus in a fluid,   b) propagating virus in said cells,   c) collecting said virus propagated in said cells,   d) completely inactivating said virus collected from said cells, and   e) purifying the inactivated virus, wherein the purification comprises ultracentrifugation or use of a sucrose gradient,   f) contacting the purified virus with a nuclease, and   g) treating said virus with detergent under conditions effective to yield a split virus,   resulting in a preparation comprising viral antigens, wherein the collected virus of step (c) is not purified prior to the inactivating step.   
     
     
         46 . The vaccine of  claim 45 , wherein said virus is an enveloped virus. 
     
     
         47 . The vaccine of  claim 46 , wherein said virus is an orthomyxo virus. 
     
     
         48 . The vaccine of  claim 46 , wherein said virus is an influenza virus. 
     
     
         49 . The vaccine of  claim 45 , wherein the concentration of non-viral protein during said inactivation is below 350 μg/ml. 
     
     
         50 . The vaccine of  claim 45 , further comprising a stabilization agent. 
     
     
         51 . The vaccine of  claim 50 , wherein said viral antigens are stabilized by addition of an effective amount of Tween 80® (polysorbate 80). 
     
     
         52 . The vaccine of  claim 51 , wherein Tween 80® is in an amount of about 0.125%. 
     
     
         53 . The vaccine of  claim 45 , further comprising a pharmaceutical carrier or an adjuvant. 
     
     
         54 . The vaccine of  claim 53 , wherein the pharmaceutical carrier is selected from the group consisting of stabilizing salts, emulgators, solubilizers, suspending agents, thickening agents and redox components. 
     
     
         55 . The vaccine of  claim 45 , wherein the vaccine is free of stabilizers. 
     
     
         56 . The vaccine of  claim 45  which comprises one or more viral antigens.