title: Single chain antibody fragment with serine protease inhibitory property capable of neutralizing toxicity of Trimeresurus mucrosquamatus venom.
abstract: Trimeresurus mucrosquamatus (TM) is one of majorities of snake envenomation with necrotic and hemorrhagic toxin in Taiwan. In this study, chickens were used as an alternative animal model for immunization with TM venom. Using phage display technology to process four rounds of panning, selected single chain variable fragments (scFv) could specifically recognize TM venom proteins, which were later identified as a group of homogeneous venom serine protease. The specific scFv antibodies showed various inhibitory effects on sheep RBC lysis induced by TM venom using an indirect hemolytic assay in vitro. In addition, the survival times of mice were extended to certain degrees when treated with these scFv antibodies individually or in a combination. To elucidate the inhibitory mechanism, we used molecular modeling to build up the serine protease structure to simulate the possible interactions with scFv antibodies. The results suggested that the CDR-loop of the scFv antibodies (3S10 or 4S1) might bind at the 99-loop of venom serine protease so as to affect substrate access due to the partial collapse of the subsite S2 and the partial movement of the subsite S4. It is hoped these chicken-derived antibodies could be applied to develop diagnostic and therapeutic agents against snakebites.
authors.affiliation: The Center of Translational Medicine, Taipei Medical University, Taipei, Taiwan; Antibody and Hybridoma Core Facility, Taipei Medical University, Taipei, Taiwan.
authors.forename: Yu-Ching
authors.initials: YC
authors.lastname: Lee
authors.affiliation: The School of Chinese Medicine for Post Baccalaureate, I-Shou University, Kaohsiung, Taiwan; Department of Chinese Medicine, E-Da Hospital, Kaohsiung, Taiwan.
authors.forename: Wang-Chuan
authors.initials: WC
authors.lastname: Chen
authors.affiliation: Antibody and Hybridoma Core Facility, Taipei Medical University, Taipei, Taiwan.
authors.forename: Meng-Huei
authors.initials: MH
authors.lastname: Liang
authors.affiliation: Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan.
authors.forename: Chi-Hsin
authors.initials: CH
authors.lastname: Lee
authors.affiliation: National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei, Taiwan.
authors.forename: Keng-Chang
authors.initials: KC
authors.lastname: Tsai
authors.affiliation: Center for Research, Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taiwan.
authors.forename: Jen-Ron
authors.initials: JR
authors.lastname: Chiang
authors.affiliation: Center for Research, Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taiwan.
authors.forename: Liao-Chun
authors.initials: LC
authors.lastname: Chiang
authors.affiliation: Department of Pathology and Laboratory Medicine, Landseed Hospital, Taoyuan, Taiwan.
authors.forename: Chi-Ching
authors.initials: CC
authors.lastname: Chen
authors.affiliation: Department of Medical Technology, Jen-Teh Junior College of Medicine, Nursing and Management, Miaoli, Taiwan.
authors.forename: Chang-Yu
authors.initials: CY
authors.lastname: Chang
authors.affiliation: Department of Medical Technology, Jen-Teh Junior College of Medicine, Nursing and Management, Miaoli, Taiwan.
authors.forename: Ching-Hsiao
authors.initials: CH
authors.lastname: Lee
authors.affiliation: Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei, Taiwan; Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan.
authors.forename: Sy-Jye
authors.initials: SJ
authors.lastname: Leu
authors.affiliation: Antibody and Hybridoma Core Facility, Taipei Medical University, Taipei, Taiwan; School of Medical Laboratory Sciences and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan; Department of Laboratory Medicine, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan. Electronic address: yangyuan@tmu.edu.tw.
authors.forename: Yi-Yuan
authors.initials: YY
authors.lastname: Yang
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D000917:Antibody Formation; D015153:Blotting, Western; D002645:Chickens; D003435:Crotalid Venoms / Q000037:antagonists & inhibitors / Q000276:immunology / Q000633:toxicity*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D062105:Molecular Docking Simulation; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D057127:Single-Chain Antibodies / Q000494:pharmacology*; D017841:Trimeresurus
chemical_list: D057134:Antibodies, Neutralizing; D003435:Crotalid Venoms; D015842:Serine Proteinase Inhibitors; D057127:Single-Chain Antibodies; C029748:Trimeresurus venoms
keywords: Molecular modeling; Phage display technology; Single chain variable fragment; Trimeresurus mucrosquamatus; Venom serine protease