title: Structural and functional characterization of a novel scFv anti-HSP60 of Strongyloides sp.
abstract: Phage display is a powerful technology that selects specific proteins or peptides to a target. We have used Phage Display to select scFv (single-chain variable fragment) clones from a combinatorial library against total proteins of Strongyloides venezuelensis. After scFv characterization, further analysis demonstrated that this recombinant fragment of antibody was able to bind to an S. venezuelensis antigenic fraction of ~65 kDa, present in the body periphery and digestive system of infective larvae (L3), as demonstrated by immunofluorescence. Mass spectrometry results followed by bioinformatics analysis showed that this antigenic fraction was a heat shock protein 60 (HSP60) of Strongyloides sp. The selected scFv was applied in serodiagnosis by immune complexes detection in serum samples from individuals with strongyloidiasis using a sandwich enzyme-linked immunosorbent assay (ELISA), showing sensitivity of 97.5% (86.84-99.94), specificity of 98.81 (93.54-99.97), positive likelihood ratio of 81.60 and an area under the curve of 0.9993 (0.9973-1.000). Our study provided a novel monoclonal scFv antibody fragment which specifically bound to HSP60 of Strongyloides sp. and was applied in the development of an innovative serodiagnosis method for the human strongyloidiasis.
authors.affiliation: Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais, Brazil.
authors.forename: Marcelo Arantes
authors.initials: MA
authors.lastname: Levenhagen
authors.affiliation: Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais, Brazil.
authors.forename: Fabiana
authors.initials: F
authors.lastname: de Almeida Araújo Santos
authors.affiliation: Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais, Brazil.
authors.forename: Patrícia Tiemi
authors.initials: PT
authors.lastname: Fujimura
authors.affiliation: Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais, Brazil.
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Carneiro
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Caneiro
authors.affiliation: Laboratório de Diagnóstico de Parasitoses, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais, Brazil.
authors.forename: Julia Maria
authors.initials: JM
authors.lastname: Costa-Cruz
authors.affiliation: 1] Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, Minas Gerais, Brazil [2] Department of Medical Microbiology and Immunology, University of California, Davis, California, United States of America.
authors.forename: Luiz Ricardo
authors.initials: LR
authors.lastname: Goulart
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000595:Amino Acid Sequence; D000818:Animals; D000947:Antigens, Helminth / Q000097:blood / Q000276:immunology*; D018834:Chaperonin 60 / Q000276:immunology*; D005260:Female; D006801:Humans; D007814:Larva / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013321:Strongyloides / Q000254:growth & development / Q000378:metabolism*; D055815:Young Adult
chemical_list: D000947:Antigens, Helminth; D018834:Chaperonin 60; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies