title: Modular protein expression by RNA trans-splicing enables flexible expression of antibody formats in mammalian cells from a dual-host phage display vector.
abstract: A recently described dual-host phage display vector that allows expression of immunoglobulin G (IgG) in mammalian cells bypasses the need for subcloning of phage display clone inserts to mammalian vectors for IgG expression in large antibody discovery and optimization campaigns. However, antibody discovery and optimization campaigns usually need different antibody formats for screening, requiring reformatting of the clones in the dual-host phage display vector to an alternative vector. We developed a modular protein expression system mediated by RNA trans-splicing to enable the expression of different antibody formats from the same phage display vector. The heavy-chain region encoded by the phage display vector is directly and precisely fused to different downstream heavy-chain sequences encoded by complementing plasmids simply by joining exons in different pre-mRNAs by trans-splicing. The modular expression system can be used to efficiently express structurally correct IgG and Fab fragments or other antibody formats from the same phage display clone in mammalian cells without clone reformatting.
authors: Department of Antibody Engineering, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors: Yonglei
authors: Y
authors: Shang
authors: Department of Antibody Engineering, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA.
authors: Devin
authors: D
authors: Tesar
authors: Department of Antibody Engineering, Genentech, 1 DNA Way, South San Francisco, CA 94080, USA ihotzel@gene.com.
authors: Isidro
authors: I
authors: Hötzel
mesh_terms: D017104:Bacteriophage M13 / Q000235:genetics; D061505:Cell Surface Display Techniques / Q000379:methods*; D005091:Exons / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000235:genetics*; D007438:Introns / Q000235:genetics; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D026723:Polyadenylation; D020040:Trans-Splicing; D014162:Transfection
chemical_list: D007074:Immunoglobulin G
keywords: bacteriophage; intron; polyadenylation; spliceosome; trans-splicing