Method for producing proteins in Pichia pastoris that lack detectable cross binding activity to antibodies against host cell antigens

US9139632B2 · kind B2 · utility

Assignee

• Merck Sharp & Dohme LLC · US

Inventors

• Piotr Bobrowicz · Lebanon, US
• Sujatha Gomathinayagam · Hanover, US
• Stephen Hamilton · Enfield, US
• Huijuan Li · Hanover, US
• Natarajan Sethuraman · Columbia, US
• Terrance A. Stadheim · White River Junction, US
• Stefan Wildt · Lebanon, US

Key dates

• Filing date: Oct 11, 2010
• Grant date: Sep 22, 2015
• Expiry date: Feb 25, 2031

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K2317/33
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Methods for producing proteins and glycoproteins in Pichia pastoris that lack detectable cross binding activity to antibodies made against host cell antigens are described. In particular, methods are described wherein recombinant Pichia pastoris strains that do not display a β-mannosyltransferase 2 activity with respect to an N-glycan or O-glycan and do not display at least one activity selected from a β-mannosyltransferase 1, 3, and 4 activity to produce recombinant proteins and glycoproteins. These recombinant Pichia pastoris strains can produce proteins and glycoproteins that lack detectable α-mannosidase resistant β-mannose residues thereon and thus, lack cross binding activity to antibodies against host cell antigens. Further described are methods for producing bi-sialylated human erythropoietin in Pichia pastoris that lack detectable cross binding activity to antibodies against host cell antigens.