title: Preparation and characterization of a human scFv against the Clostridium perfringens type A alpha-toxin.
abstract: Alpha-toxin produced by Clostridium perfringens is an important virulence factor, causing food poisoning and gas gangrene in humans. As such, it is considered a potential bioterrorism threat. To date, there is still no human effective therapeutic drug against alpha-toxin. In this study, a human single chain antibody against alpha-toxin was produced from synthetic (Tomlinson I + J) naive phage display libraries, and its preventive and therapeutic efficacy in mice was examined. To prove the neutralizing potential of the scFv, alpha-toxin was preincubated with scFv and subsequently tested for its lecithinase and hemolytic activity, as well as its lethal effect in mice following intravenous administration. The equilibrium association constant between scFv and CPA was 2.02 × 1010(1/M), as analyzed by SPR. The scFv could inhibit lecithinase and hemolytic activity, and provided effective protection against alpha-toxin when mice were challenged 1-h post scFv injection. In addition, the survival rate reached 80% for mice treated with scFv within 30 min of being challenged with a 2 × LD50 dose of alpha-toxin. These results confirmed that we successfully prepared a human scFv against C. perfringens type A alpha-toxin, which can be used in the prevention and treatment of alpha-toxin-related illness.
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, Jilin Province, China.
authors.forename: Dongdong
authors.initials: D
authors.lastname: Wang
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, Jilin Province, China.
authors.forename: Yuhuan
authors.initials: Y
authors.lastname: Yue
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, Jilin Province, China.
authors.forename: Guangmou
authors.initials: G
authors.lastname: Wu
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, Jilin Province, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, Jilin Province, China.
authors.forename: Yuling
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Graduate Academy, Jilin Agriculture University, Changchun, 130118, Jilin Province, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Yu
authors.affiliation: Graduate Academy, Jilin Agriculture University, Changchun, 130118, Jilin Province, China.
authors.forename: Yuangang
authors.initials: Y
authors.lastname: Ji
authors.affiliation: Graduate Academy, Jilin University, Changchun, 130062, Jilin Province, China.
authors.forename: Jinpeng
authors.initials: J
authors.lastname: Wang
authors.affiliation: Graduate Academy, Jilin University, Changchun, 130062, Jilin Province, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Li
authors.affiliation: Graduate Academy, Yangzhou University, Yangzhou, 225009, Jiangsu Province, China.
authors.forename: Rongrong
authors.initials: R
authors.lastname: Pan
authors.affiliation: Graduate Academy, Jilin Agriculture University, Changchun, 130118, Jilin Province, China.
authors.forename: Hongyuan
authors.initials: H
authors.lastname: Ma
authors.affiliation: Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun, 130122, Jilin Province, China. Electronic address: zhangguoli2001@126.com.
authors.forename: Guoli
authors.initials: G
authors.lastname: Zhang
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000276:immunology* / Q000633:toxicity; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000276:immunology* / Q000633:toxicity; D006801:Humans; D007928:Lethal Dose 50; D051379:Mice; D019151:Peptide Library; D017422:Sequence Analysis, DNA; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000302:isolation & purification / Q000627:therapeutic use*; D010738:Type C Phospholipases / Q000276:immunology* / Q000633:toxicity
chemical_list: D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D019151:Peptide Library; D057127:Single-Chain Antibodies; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens
keywords: Alpha toxin; Biological activity; Biopanning; Clostridium perfringens type A; Human single chain antibody