title: Recognition of Bungarus multicinctus venom by a DNA aptamer against β-bungarotoxin.
abstract: Antibody-based technology is the main method for diagnosis and treatment of snake bite envenoming currently. However, the development of an antibody, polyclonal or monoclonal, is a complicated and costly procedure. Aptamers are single stranded oligonucleotides that recognize specific targets such as proteins and have shown great potential over the years as diagnostic and therapeutic agents. In contrast to antibodies, aptamers can be selected in vitro without immunization of animals, and synthesized chemically with extreme accuracy, low cost and high degree of purity. In this study we firstly report on the identification of DNA aptamers that bind to β-bungarotoxin (β-BuTx), a neurotoxin from the venom of Bungarus multicinctus. A plate-SELEX method was used for the selection of β-BuTx specific aptamers. After 10 rounds of selection, four aptamer candidates were obtained, with the dissociation constant ranged from 65.9 nM to 995 nM measured by fluorescence spectroscopy. Competitive binding assays using both the fluorescently labeled and unlabeled aptamers revealed that the four aptamers bound to the same binding site of β-BuTx. The best binder, βB-1, bound specifically to β-BuTx, but not to BSA, casein or α-Bungarotoxin. Moreover, electrophoretic mobility shift assay and enzyme-linked aptamer assay demonstrated that βB-1 could discriminate B. multicinctus venom from other snake venoms tested. The results suggest that aptamer βB-1 can serve as a useful tool for the design and development of drugs and diagnostic tests for β-BuTx poisoning and B. multicinctus bites.
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing, China; Institute of Military Medicine, Chengdu Military Region's Center for Disease Control & Prevention, Kunming, China.
authors.forename: Fengping
authors.initials: F
authors.lastname: Ye
authors.affiliation: Institute of Military Medicine, Chengdu Military Region's Center for Disease Control & Prevention, Kunming, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Institute of Military Medicine, Chengdu Military Region's Center for Disease Control & Prevention, Kunming, China.
authors.forename: Xi
authors.initials: X
authors.lastname: Wang
authors.affiliation: Institute of Military Medicine, Chengdu Military Region's Center for Disease Control & Prevention, Kunming, China.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Tan
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing, China.
authors.forename: Wenwen
authors.initials: W
authors.lastname: Xin
authors.affiliation: Institute of Military Medicine, Chengdu Military Region's Center for Disease Control & Prevention, Kunming, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.affiliation: Institute of Military Medicine, Chengdu Military Region's Center for Disease Control & Prevention, Kunming, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Institute of Military Medicine, Chengdu Military Region's Center for Disease Control & Prevention, Kunming, China.
authors.forename: Quanshui
authors.initials: Q
authors.lastname: Fan
authors.affiliation: State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Beijing, China.
authors.forename: Jinglin
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001483:Base Sequence; D001665:Binding Sites; D002038:Bungarotoxins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D017838:Bungarus / Q000378:metabolism; D052156:SELEX Aptamer Technique; D012910:Snake Venoms / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D052157:Aptamers, Nucleotide; D002038:Bungarotoxins; D012910:Snake Venoms