title: Production of scFv-conjugated affinity silk film and its application to a novel enzyme-linked immunosorbent assay.
abstract: Bombyx mori (silkworm) silk proteins have been utilized as unique biomaterials for various medical applications. To develop a novel affinity silk material, we generated a transgenic silkworm that spins silk protein containing the fibroin L-chain linked with the single-chain variable fragment (scFv) as a fusion protein. Previously, the scFv-conjugated "affinity" silk powder specifically immunoprecipitated its target protein, Wiskott-Aldrich syndrome protein. To expand the applicability of affinity silk materials, we processed the scFv-conjugated silk protein into a thin film by dissolving it in lithium bromide, then drying it in the wells of 96-well plates. Enzyme-linked immunosorbent assay demonstrated specific detection of Wiskott-Aldrich syndrome protein, both as a recombinant protein and in its native form extracted from mouse macrophages. These findings suggest that this scFv-conjugated silk film serves as the basis for an alternative immunodetection system.
authors: 1] Animal Immune and Cell Biology Research Unit, National Institute of Agrobiological Sciences, 1-2 Ohwashi, Tsukuba, Ibaraki 305-8634, Japan [2].
authors: Mitsuru
authors: M
authors: Sato
authors: 1] Silk Materials Research Unit, National Institute of Agrobiological Sciences, 1-2 Ohwashi, Tsukuba, Ibaraki 305-8634, Japan [2].
authors: Katsura
authors: K
authors: Kojima
authors: Animal Immune and Cell Biology Research Unit, National Institute of Agrobiological Sciences, 1-2 Ohwashi, Tsukuba, Ibaraki 305-8634, Japan.
authors: Chisato
authors: C
authors: Sakuma
authors: Silk Materials Research Unit, National Institute of Agrobiological Sciences, 1-2 Ohwashi, Tsukuba, Ibaraki 305-8634, Japan.
authors: Maria
authors: M
authors: Murakami
authors: Silk Materials Research Unit, National Institute of Agrobiological Sciences, 1-2 Ohwashi, Tsukuba, Ibaraki 305-8634, Japan.
authors: Yasushi
authors: Y
authors: Tamada
authors: Animal Immune and Cell Biology Research Unit, National Institute of Agrobiological Sciences, 1-2 Ohwashi, Tsukuba, Ibaraki 305-8634, Japan.
authors: Hiroshi
authors: H
authors: Kitani
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry / Q000276:immunology; D012831:Bombyx / Q000378:metabolism; D001965:Bromides / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D047468:Immunoprecipitation; D018020:Lithium Compounds / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D047011:Silk / Q000235:genetics / Q000378:metabolism*; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051316:Wiskott-Aldrich Syndrome Protein / Q000032:analysis / Q000276:immunology
chemical_list: D055910:Antibodies, Immobilized; D001965:Bromides; D018020:Lithium Compounds; D011993:Recombinant Fusion Proteins; D047011:Silk; D057127:Single-Chain Antibodies; D051316:Wiskott-Aldrich Syndrome Protein; C040949:lithium bromide