title: Functional stability of 3D8 scFv, a nucleic acid-hydrolyzing single chain antibody, under different biochemical and physical conditions.
abstract: 3D8 single-chain Fv (scFv) is a catalytic nucleic acid antibody with anti-viral activity against a broad spectrum of viruses. Here we investigated the functional stability of 3D8 scFv to provide a basis for engineering a 3D8 scFv derivative and for developing stable formulations with improved stability and potential use as an anti-viral agent. The stability of 3D8 scFv was assessed by measuring its DNA-hydrolyzing activity under different biochemical and physical conditions using a fluorescence resonance energy transfer (FRET)-based method. In addition, the anti-influenza (H9N2) effect of 3D8 scFv was evaluated in A549 cells. 3D8 scFv was stable at 50°C for 6h at pH 7.2, for 3 days at pH 4-10 at 37°C and 30 days at pH 4-8 at 37°C. The stability was not affected by a reducing condition, freeze-thawing for up to 30 cycles, or lyophilization. Evaluation of the anti-virus effect showed that cells treated with 32-128 units of 3D8 scFv showed a 50% decrease in influenza replication compared to untreated cells. Based on its enzymatic stability in various biochemical and physical environments, 3D8 scFv holds good potential for development as an anti-viral therapeutic.
authors.affiliation: Department of Biomedical Sciences, Graduate School, Ajou University, 206 World Cup-ro, Yeongtong-gu, Suwon 443-749, Republic of Korea.
authors.forename: Joungmin
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Biomedical Sciences, Graduate School, Ajou University, 206 World Cup-ro, Yeongtong-gu, Suwon 443-749, Republic of Korea.
authors.forename: Hyunjoon
authors.initials: H
authors.lastname: Park
authors.affiliation: Department of Biomedical Sciences, Graduate School, Ajou University, 206 World Cup-ro, Yeongtong-gu, Suwon 443-749, Republic of Korea.
authors.forename: Minjae
authors.initials: M
authors.lastname: Kim
authors.affiliation: Department of Biomedical Sciences, Graduate School, Ajou University, 206 World Cup-ro, Yeongtong-gu, Suwon 443-749, Republic of Korea.
authors.forename: Youngsil
authors.initials: Y
authors.lastname: Seo
authors.affiliation: Department of Biomedical Sciences, Graduate School, Ajou University, 206 World Cup-ro, Yeongtong-gu, Suwon 443-749, Republic of Korea.
authors.forename: Yeonjin
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Animal Biotechnology Division, National Institute of Animal Science, Rural Development Administration, Suwon 441-706, Republic of Korea.
authors.forename: Sung June
authors.initials: SJ
authors.lastname: Byun
authors.affiliation: Department of Genetic Engineering, Sungkyunkwan University, Jangan-gu, Suwon, Republic of Korea.
authors.forename: Sukchan
authors.initials: S
authors.lastname: Lee
authors.affiliation: Department of Biomedical Sciences, Graduate School, Ajou University, 206 World Cup-ro, Yeongtong-gu, Suwon 443-749, Republic of Korea; Department of Microbiology, Ajou University School of Medicine, 206 World Cup-ro, Yeongtong-gu, Suwon 443-749, Republic of Korea. Electronic address: kwonmh@ajou.ac.kr.
authors.forename: Myung-Hee
authors.initials: MH
authors.lastname: Kwon
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D000998:Antiviral Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004355:Drug Stability; D006367:HeLa Cells; D006801:Humans; D006868:Hydrolysis; D053127:Influenza A Virus, H9N2 Subtype / Q000187:drug effects* / Q000378:metabolism; D009696:Nucleic Acids / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology
chemical_list: D000911:Antibodies, Monoclonal; D000998:Antiviral Agents; D009696:Nucleic Acids; D057127:Single-Chain Antibodies
keywords: Anti- influenza activity; Anti-nucleic acid antibody; Antibody stability; Catalytic antibody; DNA hydrolysis