title: Improving the developability of an anti-EphA2 single-chain variable fragment for nanoparticle targeting.
abstract: Antibody-targeted nanoparticles have great promise as anti-cancer drugs; however, substantial developmental challenges of antibody modules prevent many candidates from reaching the clinic. Here, we describe a robust strategy for developing an EphA2-targeting antibody fragment for immunoliposomal drug delivery. A highly bioactive single-chain variable fragment (scFv) was engineered to overcome developmental liabilities, including low thermostability and weak binding to affinity purification resins. Improved thermostability was achieved by modifying the framework of the scFv, and complementarity-determining region (CDR)-H2 was modified to increase binding to protein A resins. The results of our engineering campaigns demonstrate that it is possible, using focused design strategies, to rapidly improve the stability and manufacturing characteristics of an antibody fragment for use as a component of a novel therapeutic construct.
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Melissa L
authors.initials: ML
authors.lastname: Geddie
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Neeraj
authors.initials: N
authors.lastname: Kohli
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Dmitri B
authors.initials: DB
authors.lastname: Kirpotin
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Maja
authors.initials: M
authors.lastname: Razlog
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Jiao
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Tad
authors.initials: T
authors.lastname: Kornaga
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Rennard
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Lihui
authors.initials: L
authors.lastname: Xu
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Birgit
authors.initials: B
authors.lastname: Schoerberl
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.|b Department of Anesthesia and Pharmaceutical Chemistry , University of California San Francisco , San Francisco , CA , USA.
authors.forename: James D
authors.initials: JD
authors.lastname: Marks
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Daryl C
authors.initials: DC
authors.lastname: Drummond
authors.affiliation: a Merrimack, Inc. , Cambridge , MA , USA.
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Lugovskoy
mesh_terms: D000818:Animals; D016503:Drug Delivery Systems / Q000379:methods*; D036383:Ephrin-A2 / Q000276:immunology*; D006801:Humans; D018796:Immunoconjugates / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology; D053758:Nanoparticles; D015202:Protein Engineering / Q000379:methods; D055550:Protein Stability; D036104:Receptor, EphA2; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000276:immunology*
chemical_list: C000623258:EPHA2 protein, human; D036383:Ephrin-A2; D018796:Immunoconjugates; D007135:Immunoglobulin Variable Region; D057127:Single-Chain Antibodies; D036104:Receptor, EphA2
keywords: Antibody-drug conjugate; antibody engineering; antibody fragment; developability; liposome; manufacturability; protein A binding; stability