title: Antigen fingerprinting of polyclonal antibodies raised in immunized chickens with tick total proteins: a reservoir for the discovery of novel antigens.
abstract: Identification of tick-protective antigens remains the limiting step in vaccine development. The authors have generated several B cell epitope candidates by fingerprinting Rhipicephalus (Boophilus) microplus proteins that were characterized through bioselection of random peptide phage display libraries against polyclonal antibodies antitick proteins. From 280 clones selected and sequenced, 107 distinct reactive clones were validated by dot-blot assays. Eight consensus motifs were generated, and the most frequent ones were PXXKXH, NXXKXXL, and HTS (68.2%, 65%, and 42%, respectively). The consensus sequences identified potential vaccine targets by alignment with the protein database of R. microplus, which may have putative roles in the host response. Sequences that did not align with known proteins but shared extensive homology among each other were classified as conformational epitopes. Sequence alignments also recognized multiple targets, and the most predominant proteins were identified. Finally, immunized mice sera recognized tick proteins, demonstrating that functional epitope profiles can be identified through selection of phage-displayed peptide libraries with hyperimmune sera and revealing that the epitope-displaying phages can be used as potential vaccine immunogens.
authors.affiliation: Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil. crprudencio@uol.com.br
authors.forename: Carlos Roberto
authors.initials: CR
authors.lastname: Prudencio
authors.forename: Aline Aparecida Rezende
authors.initials: AA
authors.lastname: Rodrigues
authors.forename: Rone
authors.initials: R
authors.lastname: Cardoso
authors.forename: Matias Pablo Juan
authors.initials: MP
authors.lastname: Szabó
authors.forename: Luiz Ricardo
authors.initials: LR
authors.lastname: Goulart
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity; D000941:Antigens / Q000737:chemistry / Q000276:immunology*; D060829:Arthropod Proteins / Q000737:chemistry / Q000276:immunology*; D002645:Chickens / Q000276:immunology*; D018604:Epitope Mapping / Q000379:methods*; D007118:Immunoassay; D051379:Mice; D008807:Mice, Inbred BALB C; D019151:Peptide Library; D011485:Protein Binding / Q000276:immunology; D048494:Rhipicephalus / Q000276:immunology*; D020539:Sequence Analysis, Protein
chemical_list: D000906:Antibodies; D000941:Antigens; D060829:Arthropod Proteins; D019151:Peptide Library