title: Antibody discovery ex vivo accelerated by the LacO/LacI regulatory network.
abstract: Monoclonal antibodies (mAbs) can be potent and highly specific therapeutics, diagnostics and research reagents. Nonetheless, mAb discovery using current in vivo or in vitro approaches can be costly and time-consuming, with no guarantee of success. We have established a platform for rapid discovery and optimization of mAbs ex vivo. This DTLacO platform derives from a chicken B cell line that has been engineered to enable rapid selection and seamless maturation of high affinity mAbs. We have validated the DTLacO platform by generation of high affinity and specific mAbs to five cell surface targets, the receptor tyrosine kinases VEGFR2 and TIE2, the glycoprotein TROP2, the small TNF receptor family member FN14, and the G protein-coupled receptor FZD10. mAb discovery is rapid and humanization is straightforward, establishing the utility of the DTLacO platform for identification of mAbs for therapeutic and other applications.
authors.affiliation: Department of Immunology, University of Washington School of Medicine, Seattle, Washington, USA.
authors.forename: Munehisa
authors.initials: M
authors.lastname: Yabuki
authors.forename: W Jason
authors.initials: WJ
authors.lastname: Cummings
authors.forename: John B
authors.initials: JB
authors.lastname: Leppard
authors.forename: Robert M
authors.initials: RM
authors.lastname: Immormino
authors.forename: Christi L
authors.initials: CL
authors.lastname: Wood
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Allison
authors.forename: Patrick W
authors.initials: PW
authors.lastname: Gray
authors.forename: Larry W
authors.initials: LW
authors.lastname: Tjoelker
authors.forename: Nancy
authors.initials: N
authors.lastname: Maizels
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry / Q000276:immunology; D000915:Antibody Affinity / Q000276:immunology; D002460:Cell Line; D002645:Chickens; D002999:Clone Cells; D022801:Complementarity Determining Regions / Q000235:genetics; D017124:Conserved Sequence / Q000235:genetics; D053263:Gene Regulatory Networks / Q000235:genetics*; D005818:Genetic Engineering; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000276:immunology; D007763:Lac Operon / Q000235:genetics*; D056968:Lac Repressors / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011956:Receptors, Cell Surface / Q000276:immunology; D019809:Streptavidin / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D022801:Complementarity Determining Regions; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D056968:Lac Repressors; D011956:Receptors, Cell Surface; D019809:Streptavidin