title: Optimizing antibody expression by using the naturally occurring framework diversity in a live bacterial antibody display system.
abstract: Rapid identification of residues that influence antibody expression and thermostability is often needed to move promising therapeutics into the clinic. To establish a method that can assess small expression differences, we developed a Bacterial Antibody Display (BAD) system that overcomes previous limitations, enabling the use of full-length formats for antibody and antigen in a live cell setting. We designed a unique library of individual framework variants using natural diversity introduced by somatic hypermutation, and screened half-antibodies for increased expression using BAD. We successfully identify variants that dramatically improve expression yields and in vitro thermostability of two therapeutically relevant antibodies in E. coli and mammalian cells. While we study antibody expression, bacterial display can now be expanded to examine the processes of protein folding and translocation. Additionally, our natural library design strategy could be applied during antibody humanization and library design for in vitro display methods to maintain expression and formulation stability.
authors.affiliation: Department of Antibody Engineering, Genentech Research and Early Development, 1 DNA Way, South San Francisco, CA, 94080, USA.
authors.forename: T Noelle
authors.initials: TN
authors.lastname: Lombana
authors.affiliation: Department of Antibody Engineering, Genentech Research and Early Development, 1 DNA Way, South San Francisco, CA, 94080, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Dillon
authors.affiliation: Department of Antibody Engineering, Genentech Research and Early Development, 1 DNA Way, South San Francisco, CA, 94080, USA.
authors.forename: Jack
authors.initials: J
authors.lastname: Bevers
authors.affiliation: Department of Antibody Engineering, Genentech Research and Early Development, 1 DNA Way, South San Francisco, CA, 94080, USA.
authors.forename: Christoph
authors.initials: C
authors.lastname: Spiess
mesh_terms: D000906:Antibodies / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D014644:Genetic Variation; D018793:Interleukin-13 / Q000276:immunology; D019151:Peptide Library; D015202:Protein Engineering / Q000379:methods*; D055550:Protein Stability; D036223:Receptor, EphB3; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000276:immunology
chemical_list: D000906:Antibodies; D018793:Interleukin-13; D019151:Peptide Library; D011994:Recombinant Proteins; D042461:Vascular Endothelial Growth Factor A; D036223:Receptor, EphB3