Crucell Holland B.v.

Janssen Vaccines & Prevention B.v.

John Alfred Lewis

Taco Gilles Uil

Barbara Petronella Sanders

Diana Edo-Matas

Alfred Luitjens

Jerome Hhv Custers

A poliovirus (PV) strain was attenuated by a novel method of Cold-Adapted-Viral-Attenuation (CAVA). The resulting recombinant attenuated PV, CAVA-PV, shows wild-type replication at 30&#xb0; C., but no substantial replication at 37&#xb0; C. The inability to replicate at 37&#xb0; C. is defined by an inability to quantify virus during infection at this temperature by titration (infectious units), qPCR (viral RNA) or Electron Microscopy (visual signs of infection). CAVA-PV is genetically stable under production conditions and shows utility for use as the backbone to produce attenuated strains with the same antigenic profile as conventional vaccines by replacing the sequence coding for the capsid of CAVA-PV with sequences coding for capsids of different PV strains. Furthermore, mutations identified in CAVA-PV can be engineered into different, even wild-type and neurovirulent poliovirus background strains to obtain additional CAVA-PV strains.

Cold-adapted-viral-attenuation (CAVA) and novel attenuated poliovirus strains

The invention provides methods for large-scale production of recombinant adenovirus 35, using perfusion systems and infection at very high-cell densities.

Method for the production of adenoviral vectors

Described is a process for producing poliovirus, the process comprising: a) providing a serum-free suspension culture of cells, which are primary human retina (HER) cells that have been immortalized by expression of adenovirus E1 sequences, b) infecting the cells with poliovirus, at a cell density of between 2&#xd7;10cells/ml and 150&#xd7;10cells/ml, and c) harvesting poliovirus at a time of between 12 and 48 hours after infection.

Production of poliovirus at high titers for vaccine production

Provided is a process for the production of poliovirus, comprising the steps of:

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Little Compton, RI, United States of America

John Alfred Lewis