Method and device for producing vaccine

Abstract

A method of making a vaccine using animal derived component free (ADCF) cell culture technology, including the steps of attaching ADCF-adapted cells to a microcarrier including an attachment mechanism for attaching filipodia of the cells, the microcarrier being in a culture, growing the cells in ADCF maintenance media, infecting the cells with vaccine media, producing virus within the cells, and harvesting the virus. A vaccine produced by the above method in a pharmaceutically acceptable carrier. A vaccine production structure of ADCF-adapted cells removably attached to microcarrier beads including an attachment mechanism for attaching filipodia of the cells.

1 - 33 . (canceled) 
     
     
         34 . A method of making an influenza vaccine including the steps of:
 selecting a cell line from the group consisting of influenza-permissive cells having attachment filipodia;   attaching said cells to a plurality of microcarriers including attachment means for attaching said filipodia of the cells;   placing said plurality of microcarriers with said attached cells in a culture;   suspending said plurality of microcarriers by an impeller at a speed sufficient to maintain said microcarriers in a substantially homogenous suspension;   growing said cells in a maintenance media;   infecting said cells with influenza vaccine media;   producing influenza virus within said cells; and   harvesting said influenza virus.   
     
     
         35 . The method of  claim 34  wherein the attaching step is further defined as removing cells from a substrate using a protease to result in a single cell suspension, adding said single cell suspension to said culture, uniformly attaching said cells to said microcarriers, and spreading said cells to said microcarriers. 
     
     
         36 . The method of  claim 34 , wherein the producing step is further defined as timing cell infection relative to cell seeding density on said microcarriers and growth phase of the cell to synergistically produce high virus yield. 
     
     
         37 . The method of  claim 34 , wherein the harvesting step is further defined as harvesting supernatant to collect said virus. 
     
     
         38 . The method of  claim 35 , further including the step of transferring said microcarriers to said culture at least one hour before adding said single cell suspension to said culture. 
     
     
         39 . The method of  claim 34 , wherein each of said microcarriers has a micro-porous surface. 
     
     
         40 . The method of  claim 39 , wherein said microcarriers have a density of 1.04 to 1.1 g/cc. 
     
     
         41 . The method of  claim 39 , wherein each of said microcarriers has a diameter of 75 to 225 micrometers. 
     
     
         42 . The method of  claim 34 , wherein said microcarriers are composed of a material selected from the group consisting of glass, polystyrene plastic, acrylamide, dextran, solid collagen, porous collagen, porcine collagen, cellulose, and liquid fluorocarbon. 
     
     
         43 . The method of  claim 34 , wherein said microcarriers are composed of a material chosen from the group consisting of porcine collagen coated polystyrene and glass-coated polystyrene. 
     
     
         44 . The method of  claim 34 , wherein said microcarriers include at least one adhesive peptide attached to a surface of said microcarriers through covalent or non-covalent linkages. 
     
     
         45 . The method of  claim 34 , wherein said microcarriers include a coating chosen from the group consisting of porcine collagen, bovine collagen, human collagen, ProNectin®, recombinant fibronectin, and any other suitable natural or synthetic peptide.