Production of low fucose antibodies in h4-ii-e rat cells

A method for producing a glycoprotein of interest comprising the steps of:

• a) providing a rat hepatoma cell,
• b) optionally adapting said cell of step a) to growth in suspension culture,
• c) optionally adapting said cell of step a) and/or step b) to growth in serum-free medium,
• d) optionally adapting said cell of step a) and/or step b) and/or step c) to growth in calcium-reduced or calcium-free medium,
• e) transfecting this optionally adapted rat hepatoma cell with a nucleic acid sequence encoding a recombinant glycoprotein of interest,
• f) cultivating said transfected cell of step e) under conditions which allow expression of said glycoprotein of interest, and
• g) optionally isolating and purifying said glycoprotein of interest.

A method for producing a (recombinant) antibody or Fc-fusion protein having

• a) FcγRIIIa binding activity and/or
• b) complement binding activity and/or
• c) binding activity of the neonatal Fc receptor FcRn,
• comprising producing said antibody or Fc fusion protein in a rat hepatoma cell, wherein said rat hepatoma cell is preferably a H4-II-E cell, or said cell is:
• i) a cell derived from a cell selected from the group consisting of:
• European Collection of Cell Cultures (ECACC, Cat. no. 87031301), American Type Culture Collection (ATCC, deposit no. CRL-1548), H4-II-E-C3 cell line (CRL-1600 or HPACC No. 85061112 or ECACC catalogue no. 85061112), H4II cell line (HPACC Nr. 89042702), H4-TG cell line (CRL-1578), H5 cell line (HPACC, Nr. 94101905) and H4-S cell line (HPACC Nr. 89102001), or
• ii) a cell which is deposited with the European Collection of Cell Cultures under the number ECACC, Cat. no. 87031301 or the American Type Culture Collection ATCC under the deposit no. CRL-1548, or
• iii) a cell which is deposited with the DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH) under the accession number DSM ACC3129 (H4-II-E), or
• iv) a cell which is deposited with the DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH) under the accession number DSM ACC3130 (H4-II-Es), or
• v) a derivative or progeny of any one cell of i) or ii) or iii) or iv).

A method of generating a host cell for production of recombinant glycoprotein comprising:

• a) providing a rat hepatoma cell,
• b) adapting said rat hepatoma cell of step a) to growth in suspension culture, and
• c) adapting said rat hepatoma cell of step a) to growth in serum-free medium, and
• d) adapting said rat hepatoma cell of step a) to growth in calcium-reduced or calcium-free medium, and
• e) optionally adapting said rat hepatoma cell of step a) to growth in medium free of any protein/peptide of animal origin,
• f) optionally selecting a single cell clone, and
• g) obtaining a host cell.