title: Recombinant human antibody single chain variable fragments reactive with Candida albicans surface antigens.
abstract: A combinatorial phage display library expressing human immunoglobulin heavy and light chain variable regions was used to identify phage clones capable of binding to the surface of Candida albicans blastoconidia. Single chain antibody variable fragments (scFv) derived from three clones detected C. albicans antigens by indirect immunofluorescence assay (IFA), enzyme-linked immunosorbent assay (ELISA), and Western blotting. The antigens detected were conserved among different strains of C. albicans and several other Candida species. Two scFv clones detected antigens specifically expressed by C. albicans blastoconidia; the third detected antigens in both blastoconidia and filamentous forms of C. albicans. The antigens containing the epitopes recognized by all three scFv could be extracted from blastoconidia by dithiothreitol, suggesting attachment to the cell wall via sulfhydryl bonds. Epitope detection by the scFv was sensitive to treatment of C. albicans blastoconidia with sodium periodate, but not proteinase K, indicating the cognate epitopes were composed of carbohydrate. Antigenic determinants for each of the three scFv were detected by immunohistochemical staining of skin sections from a model of cutaneous candidiasis, demonstrating expression in vivo. Through selection for the ability to bind intact organisms, the phage display system provides a means to rapidly identify monoclonal binding ligands to Candida surface antigens. Being entirely human, mature antibodies generated from the scFv have potential utility in the treatment of candidiasis.
authors.affiliation: Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, 601 Elmwood Ave, Rochester, NY 14642, USA. haid@mail.rochester.edu
authors.forename: C G
authors.initials: CG
authors.lastname: Haidaris
authors.forename: J
authors.initials: J
authors.lastname: Malone
authors.forename: L A
authors.initials: LA
authors.lastname: Sherrill
authors.forename: J M
authors.initials: JM
authors.lastname: Bliss
authors.forename: A A
authors.initials: AA
authors.lastname: Gaspari
authors.forename: R A
authors.initials: RA
authors.lastname: Insel
authors.forename: M A
authors.initials: MA
authors.lastname: Sullivan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000946:Antigens, Fungal / Q000235:genetics / Q000276:immunology*; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D002176:Candida albicans / Q000235:genetics / Q000276:immunology*; D002179:Candidiasis, Cutaneous / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000235:genetics / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D019084:Fluorescent Antibody Technique, Indirect; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D000946:Antigens, Fungal; D000954:Antigens, Surface; D017931:DNA Primers; D000939:Epitopes; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011994:Recombinant Proteins