Protein expression from multiple nucleic acids

US8771988B2 · kind B2 · utility

Assignee

• Hoffmann-La Roche Inc. · US

Inventors

• Ulrich Goepfert · Penzberg, DE
• Hendrik Knoetgen · Penzberg, DE
• Erhard Kopetzki · Penzberg, DE
• Anne Stern · Penzberg, DE

Key dates

• Filing date: Oct 9, 2008
• Grant date: Jul 8, 2014
• Expiry date: Nov 20, 2029

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K2317/515
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The current invention reports a method for the recombinant production of a secreted heterologous immunoglobulin in a CHO cell comprising the following steps: i) providing a CHO cell, which is adapted to growth in suspension culture, adapted to growth in serum-free medium, mycoplasma free, and virus free, ii) providing a vector comprising a prokaryotic origin of replication, a first nucleic acid conferring resistance to a prokaryotic selection agent, a second nucleic acid encoding the heavy chain of said heterologous immunoglobulin, a third nucleic acid encoding the light chain of said heterologous immunoglobulin, a fourth nucleic acid conferring resistance to a eukaryotic selection agent, iii) transfecting said CHO cell, wherein said transfecting comprises a) transfecting said CHO cell with said vector comprising a fourth nucleic acid conferring resistance to a first eukaryotic selection agent, b) selecting a CHO cell by growth in cultivation medium containing said first eukaryotic selection agent, c) transfecting said selected CHO cell with said vector comprising a fourth nucleic acid conferring resistance to a second eukaryotic selection agent different to said first eukaryotic s…