Method and device for producing vaccine
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. A method of making an influenza vaccine using animal derived component free (ADCF) cell culture technology, including the steps of: attaching ADCF-adapted cells to a microcarrier including attachment means for attaching filipodia of the cells and suspending the microcarrier by an impeller turning at a constant speed of 50 to 60 rpm, the microcarrier being in a culture; growing the cells in ADCF maintenance media; infecting the cells with influenza vaccine media; producing influenza virus within the cells; and harvesting the influenza virus.
2. The method of claim 1 , wherein the attaching step is further defined as removing ADCF-adapted cells from substrate using ADCF trypsin resulting in a single cell suspension, adding the single cell suspension to the culture, uniformly attaching the cells to the microcarrier, and spreading the cells to the microcarrier.
3. The method of claim 2 , wherein the spreading step is further defined as setting the pH at 7.2-7.4, setting an intermittent stirring cycle for up to 24 hours.
4. The method of claim 1 , wherein the producing step is further defined as timing cell infection relative to cell seeding density on the microcarrier and growth phase of the cell to synergistically produce high virus yield.
5. The method of claim 1 , wherein the harvesting step is further defined as harvesting the supernatant to collect the virus.
6. The method of claim 2 , further including the step of transferring the microcarrier to the culture at least one hour before adding the single cell suspension to the culture.
7. The method of claim 6 , wherein the culture includes an impeller running at a rate of 50 to 60 rpm, with a pH at 7.5 to 7.6, and temperature at 37 degrees C.
8. The method of claim 7 , wherein the impeller runs for at least 30 minutes to allow for cell attachment to the microcarrier.
9. The method of claim 1 , wherein the attachment means of the microcarrier beads is a micro-porous surface.
10. The method of claim 9 , wherein the microcarrier beads have a density of 1.04 to 1.1 g/cc.
11. The method of claim 9 , wherein the microcarrier beads have a diameter of 75 to 225 micrometers.
12. The method of claim 9 , wherein the microcarrier beads are made of a material chosen from the group consisting of glass, polystyrene plastic, acrylamide, dextran, solid collagen, porous collagen, porcine collagen, cellulose, and liquid fluorocarbon.
13. The method of claim 9 , wherein the microcarrier beads are made of a material chosen from the group consisting of porcine collagen coated polystyrene and glass-coated polystyrene.
14. The method of claim 9 , wherein the microcarrier beads include at least one adhesive peptide attached to a surface of the microcarrier bead through covalent or noncovalent linkages.
15. The method of claim 9 , wherein the microcarrier beads include a coating chosen from the group consisting of porcine, collagen, bovine collagen, human collagen, ProNectin F, recombinant fibronectin, and any other suitable natural or synthetic peptide.
16. The method of claim 1 , wherein the vaccine media is Vero cell adapted influenza virus working seeds.
17. The method of claim 1 , wherein the ADCF-adapted cells are Vero or Madin-Darby Canine Kidney (MDCK) cells.