title: Identification and characterization of B-cell epitopes of 3FTx and PLA(2) toxins from Micrurus corallinus snake venom.
abstract: The main goal of this work was to develop a strategy to identify B-cell epitopes on four different three finger toxins (3FTX) and one phospholipase A2 (PLA2) from Micrurus corallinus snake venom. 3FTx and PLA2 are highly abundant components in Elapidic venoms and are the major responsibles for the toxicity observed in envenomation by coral snakes. Overlapping peptides from the sequence of each toxin were prepared by SPOT method and three different anti-elapidic sera were used to map the epitopes. After immunogenicity analysis of the spot-reactive peptides by EPITOPIA, a computational method, nine sequences from the five toxins were chemically synthesized and antigenically and immunogenically characterized. All the peptides were used together as immunogens in rabbits, delivered with Freund's adjuvant for a first cycle of immunization and Montanide in the second. A good antibody response against individual synthetic peptides and M. corallinus venom was achieved. Anti-peptide IgGs were also cross-reactive against Micrurus frontalis and Micrurus lemniscatus crude venoms. In addition, anti-peptide IgGs inhibits the lethal and phospholipasic activities of M. corallinus crude venom. Our results provide a rational basis to the identification of neutralizing epitopes on coral snake toxins and show that their corresponding synthetic peptides could improve the generation of immuno-therapeutics. The use of synthetic peptide for immunization is a reasonable approach, since it enables poly-specificity, low risk of toxic effects and large scale production.
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: K L
authors.initials: KL
authors.lastname: Castro
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: C G
authors.initials: CG
authors.lastname: Duarte
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil, 1500, 05503-900, São Paulo, SP, Brazil.
authors.forename: H R
authors.initials: HR
authors.lastname: Ramos
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: R A
authors.initials: RA
authors.lastname: Machado de Avila
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: F S
authors.initials: FS
authors.lastname: Schneider
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: D
authors.initials: D
authors.lastname: Oliveira
authors.affiliation: Fundação Ezequiel Dias, Belo Horizonte, Brazil.
authors.forename: C F
authors.initials: CF
authors.lastname: Freitas
authors.affiliation: Departamento de Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
authors.forename: E
authors.initials: E
authors.lastname: Kalapothakis
authors.affiliation: Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil, 1500, 05503-900, São Paulo, SP, Brazil.
authors.forename: P L
authors.initials: PL
authors.lastname: Ho
authors.affiliation: Departamento de Bioquímica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil. Electronic address: olortegi@icb.ufmg.br.
authors.forename: C
authors.initials: C
authors.lastname: Chávez-Olortegui
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D001938:Brazil; D060326:Chemistry Techniques, Synthetic; D004546:Elapid Venoms / Q000737:chemistry*; D017815:Elapidae; D004797:Enzyme-Linked Immunosorbent Assay; D018985:Epitopes, B-Lymphocyte / Q000235:genetics*; D007074:Immunoglobulin G / Q000378:metabolism; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010455:Peptides / Q000235:genetics / Q000276:immunology; D054467:Phospholipases A2 / Q000235:genetics*; D014118:Toxins, Biological / Q000235:genetics*
chemical_list: D004546:Elapid Venoms; D018985:Epitopes, B-Lymphocyte; D007074:Immunoglobulin G; D010455:Peptides; D014118:Toxins, Biological; C053972:micrurus venom; D054467:Phospholipases A2
keywords: Antivenoms; Epitopes; Micrurus corallinus snake venom; Phospholipase A(2) (PLA(2)); Synthetic peptides; Three-finger toxins (3FTx)