Development of a high yielding E. coli periplasmic expression system for the production of humanized Fab' fragments

• Published in: Biotechnology progress Vol. 33; no. 1; pp. 212 - 220
• Main Authors: Ellis, Mark, Patel, Pareshkumar, Edon, Marjory, Ramage, Walter, Dickinson, Robert, Humphreys, David P.
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.01.2017
• Subjects: Antibodies, Monoclonal, Humanized - biosynthesis; Antibodies, Monoclonal, Humanized - chemistry; Antibodies, Monoclonal, Humanized - isolation & purification; DsbC; E coli; Escherichia coli; Escherichia coli - chemistry; Escherichia coli - genetics; Fab; Fermentation; Humans; Immunoglobulin Fab Fragments - biosynthesis; Immunoglobulin Fab Fragments - chemistry; Immunoglobulin Fab Fragments - isolation & purification; Mutation; Peptide Hydrolases - chemistry; Peptide Hydrolases - isolation & purification; Periplasm - chemistry; Periplasm - genetics; Protein Engineering; Protein Folding; Tsp; DsbC; Tsp; fermentation; Escherichia coli; Fab

Humanized Fab′ fragments may be produced in the periplasm of Escherichia coli but can be subject to degradation by host cell proteases. In order to increase Fab′ yield and reduce proteolysis we developed periplasmic protease deficient strains of E. coli. These strains lacked the protease activity of...