Solohill Engineering, Inc.

University of Michigan

Other

William J Hillegas

David E Solomon

James Varani

Gilbert H Wuttke

David L Melmoth

Bonnie L Wallace

David L Helmreich

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.

Method and device for producing vaccine

There is provided a method of making microcarrier beads having the steps of forming a bead made of a lightly crosslinked styrene copolymer core and also having functional groups on the surface of the bead and washing the microcarrier beads with basic and acidic solutions to make the beads compatible for cell culture. Also provided is a microcarrier bead made of a styrene copolymer core with a tri-methylamine exterior which has been washed in basic and acidic solutions to make the beads compatible for cell culture. The method of using microcarrier beads for increased growth of anchorage dependent cells having the steps of washing the microcarrier bead with basic and acidic solutions and mixing the microcarrier bead with an anchorage dependent cell containing culture medium is also provided.

Microcarrier beads having a styrene copolymer core and a covalently linked tri-methylamine exterior

A microcarrier bead system for culturing anchorage-dependent cells is formed of a polystyrene core with a coating of collagen fixed thereover. In certain embodiments, the coating is a protein, such as laminin or fibronectin. The microcarrier bead is of low density, illustratively 1.02 g/cc, and therefore requires less agitation of the nutrient media to maintain suspension. This reduced stirring causes lower shear forces to impinge upon the cells, thereby improving the attachment and proliferation of the cells being cultured. The microcarrier bead of the present invention exhibits surprising advantages with respect to cell attachment and harvesting over beads formed entirely of collagen, or of DEAE-dextran coated with collagen. During harvesting, contamination of the product resulting from dissolved collagen, particularly when proteolytic enzymes are used, is minimized. Additionally, adsorption of toxins and product by the subject microcarrier beads is minimized.

Collagen-coated polystyrene microcarrier beads

A filter-separator pour-out cap for attachment to the neck opening of a container to separate particulate material from a liquidic suspension in the container when the container is tipped to pour out its contents. The cap comprises a filter-separator section having an upstream face at which flow from the container enters and a downstream face at which flow exits. This section comprises successive elements that are spaced axially apart from each other and are constructed and arranged to present at the upstream face juxtaposed uniformally spaced apart edges that define the entrances of openings that pass through the filter-separator from the upstream face to the downstream face. These elements have, in transverse cross section, a taper which narrows in the direction from the upstream face to the downstream face so that the openings are of progressively increasing taper in the same direction. In one embodiment of the invention these elements are identical circular rings. In another, the elements are convolutions of a helix. Various embodiments of caps are disclosed.

Filter-separator pour-out cap

An insert for a roller bottle of the type used in cell culture for vaccine manufacture is formed of a polymeric resilient material whereby it can be bent or rolled so that it can pass through the opening of the roller bottle. Once the insert is inside the bottle, it is released so that its resilient characteristic causes it to unbend or unroll. The insert is wider than the diameter of the roller bottle, and therefore it is urged against the interior wall of the roller bottle. Apertures and/or notches are provided in the insert to increase the turbulence of a fluid and to decrease the fluidic shear forces in the roller bottle as it is rotated in conventional operation. A significant increase in productivity is achieved because particles, which may be microcarrier beads, are maintained in suspension, and prevented from sinking to the bottom of the fluid medium where cells cannot grow. The roller bottle system is thereby transformed into a microcarrier suspension culture system.

Growing community of inventors

Ann Arbor, MI, United States of America

William J Hillegas