title: Functional consequences of complementarity-determining region deactivation in a multifunctional anti-nucleic acid antibody.
abstract: Many murine monoclonal anti-DNA antibodies (Abs) derived from mice models for systemic lupus erythematosus have additional cell-penetration and/or nucleic acid-hydrolysis properties. Here, we examined the influence of deactivating each complementarity-determining region (CDR) within a multifunctional anti-nucleic acid antibody (Ab) that possesses these activities, the catalytic 3D8 single chain variable fragment (scFv). CDR-deactivated 3D8 scFv variants were generated by replacing all of the amino acids within each CDR with Gly/Ser residues. The structure of 3D8 scFv accommodated single complete CDR deactivations. Different functional activities of 3D8 scFv were affected differently depending on which CDR was deactivated. The only exception was CDR1, located within the light chain (LCDR1); deactivation of LCDR1 abolished all of the functional activities of 3D8 scFv. A hybrid Ab, HW6/3D8L1, in which the LCDR1 from an unrelated Ab (HW6) was replaced with the LCDR1 from 3D8, acquired all activities associated with the 3D8 scFv. These results suggest that the activity of a multifunctional 3D8 scFv Ab can be modulated by single complete CDR deactivation and that the LCDR1 plays a crucial role in maintaining Ab properties. This study presents a new approach for determining the role of individual CDRs in multifunctional Abs with important implications for the future of Ab engineering.
authors.affiliation: From the Department of Biomedical Sciences, Graduate School, Ajou University, San 5, Woncheon-dong, Yeongtong-gu, Suwon 443-749, South Korea and.
authors.forename: Jiyeon
authors.initials: J
authors.lastname: Lee
authors.forename: Hye-Jin
authors.initials: HJ
authors.lastname: Kim
authors.forename: Jooho
authors.initials: J
authors.lastname: Roh
authors.forename: Youngsil
authors.initials: Y
authors.lastname: Seo
authors.forename: Minjae
authors.initials: M
authors.lastname: Kim
authors.forename: Hye-Ryeong
authors.initials: HR
authors.lastname: Jun
authors.forename: Chuong D
authors.initials: CD
authors.lastname: Pham
authors.forename: Myung-Hee
authors.initials: MH
authors.lastname: Kwon
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D022801:Complementarity Determining Regions / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D004247:DNA / Q000276:immunology*; D006367:HeLa Cells; D006493:Heparin / Q000378:metabolism; D006801:Humans; D006868:Hydrolysis; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D017433:Protein Structure, Secondary; D012313:RNA / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D022801:Complementarity Determining Regions; D007147:Immunoglobulin Light Chains; D057127:Single-Chain Antibodies; D012313:RNA; D006493:Heparin; D004247:DNA
keywords: Anti-DNA Antibody; Antibodies; Antibody Engineering; Autoimmunity; CDR Deactivation; Cell Penetration; DNA Hydrolysis; Immunology; Molecular Biology; Multifunctional Antibody