title: Discovery of highly soluble antibodies prior to purification using affinity-capture self-interaction nanoparticle spectroscopy.
abstract: Self-association of monoclonal antibodies (mAbs) at high concentrations can result in developability challenges such as poor solubility, aggregation, opalescence and high viscosity. There is a significant unmet need for methods that can evaluate self-association propensities of concentrated mAbs at the earliest stages in antibody discovery to avoid downstream issues. We have previously developed a method (affinity-capture self-interaction nanoparticle spectroscopy, AC-SINS) that is capable of detecting weak antibody self-interactions using unusually dilute mAb solutions (tens of µg/ml). Here we optimize and implement this assay for characterization of unpurified and highly dilute mAbs directly in cell culture media. This assay was applied to screen 87 mAbs obtained via immunization. Our measurements reveal a wide range of self-associative propensities for mAbs that bind to the same antigen and which differ mainly in their complementarity-determining regions. The least associative mAbs identified by AC-SINS were confirmed to be highly soluble when purified and concentrated by three to five orders of magnitude. This approach represents a key advance in screening mAb variants using unpurified antibody samples, and it holds significant potential to both improve initial candidate selection as well as to guide protein engineering efforts to improve the properties of specific mAb candidates.
authors.affiliation: Isermann Department of Chemical and Biological Engineering, Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Jiemin
authors.initials: J
authors.lastname: Wu
authors.affiliation: Eli Lilly Biotechnology Center, San Diego, CA 92121, USA.
authors.forename: Jason S
authors.initials: JS
authors.lastname: Schultz
authors.affiliation: Eli Lilly Biotechnology Center, San Diego, CA 92121, USA.
authors.forename: Caroline L
authors.initials: CL
authors.lastname: Weldon
authors.affiliation: Isermann Department of Chemical and Biological Engineering, Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA Present address: Protein Pharmaceutical Development, Biogen Inc., Cambridge, MA 02142, USA.
authors.forename: Shantanu V
authors.initials: SV
authors.lastname: Sule
authors.affiliation: Eli Lilly Biotechnology Center, San Diego, CA 92121, USA.
authors.forename: Qing
authors.initials: Q
authors.lastname: Chai
authors.affiliation: Isermann Department of Chemical and Biological Engineering, Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
authors.forename: Steven B
authors.initials: SB
authors.lastname: Geng
authors.affiliation: Eli Lilly Biotechnology Center, San Diego, CA 92121, USA tessier@rpi.edu cdickinson@lilly.com.
authors.forename: Craig D
authors.initials: CD
authors.lastname: Dickinson
authors.affiliation: Isermann Department of Chemical and Biological Engineering, Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY 12180, USA tessier@rpi.edu cdickinson@lilly.com.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Tessier
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification*; D005591:Chemical Fractionation / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D007114:Immunization; D053758:Nanoparticles / Q000737:chemistry*; D012995:Solubility; D013057:Spectrum Analysis
chemical_list: D000911:Antibodies, Monoclonal
keywords: aggregation; high-throughput; mAb; self-association; solubility; viscosity