title: Retrocyte Display® technology: generation and screening of a high diversity cellular antibody library.
abstract: Over the last nearly three decades in vitro display technologies have played an important role in the discovery and optimization of antibodies and other proteins for therapeutic applications. Here we describe the use of retroviral expression technology for the display of full-length IgG on B lineage cells in vitro with a hallmark of a tight and stable genotype to phenotype coupling. We describe the creation of a high-diversity (>1.0E09 different heavy- and light-chain combinations) cell displayed fully human antibody library from healthy donor-derived heavy- and light-chain gene libraries, and demonstrate the recovery of high affinity target-specific antibodies from this library by staining of cells with a labeled target antigen and their magnetic- and flow cytometry-based cell sorting. The present technology represents a further evolution in the discovery of full-length, fully human antibodies using mammalian display, and is termed Retrocyte Display® (Retroviral B lymphocyte Display).
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Breous-Nystrom
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Kornelia
authors.initials: K
authors.lastname: Schultze
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Marco
authors.initials: M
authors.lastname: Meier
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Lukas
authors.initials: L
authors.lastname: Flueck
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Christina
authors.initials: C
authors.lastname: Holzer
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Melanie
authors.initials: M
authors.lastname: Boll
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Volker
authors.initials: V
authors.lastname: Seibert
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Andrea
authors.initials: A
authors.lastname: Schuster
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Milan
authors.initials: M
authors.lastname: Blanusa
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Verena
authors.initials: V
authors.lastname: Schaefer
authors.affiliation: NBE Therapeutics AG, Basel, Switzerland.
authors.forename: Ulf
authors.initials: U
authors.lastname: Grawunder
authors.affiliation: 4-Antibody AG, Basel, Switzerland.
authors.forename: Luis
authors.initials: L
authors.lastname: Martin-Parras
authors.affiliation: 4-Antibody AG, Basel, Switzerland. Electronic address: Marc.vandijk@4-antibody.com.
authors.forename: Marc A
authors.initials: MA
authors.lastname: van Dijk
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D001402:B-Lymphocytes / Q000378:metabolism; D015925:Cryopreservation; D004353:Drug Evaluation, Preclinical; D005434:Flow Cytometry; D014644:Genetic Variation; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000235:genetics; D018189:Immunomagnetic Separation; D019151:Peptide Library; D011485:Protein Binding; D012190:Retroviridae / Q000235:genetics*
chemical_list: D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D019151:Peptide Library
keywords: A-MuLV; Abelson murine leukemia virus; B cell receptor; BCR; BiP; CDR; DR6; FACS; FCS; Flow cytometry; Fully human antibody; IRES; MACS; Mammalian cell surface antibody display; Retrocyte Display®; Retroviral B lymphocyte Display; Retroviral transduction; TNF-α; VH; Vκ; binding immunoglobulin protein; complementarity determining region; death receptor 6; fetal calf serum; fluorescence activated cell sorting; heavy chain; internal ribosome entry site; kappa light-chain; magnetic activated cell sorting; tumor necrosis factor-alpha