title: Production of scFv-conjugated affinity silk powder by transgenic silkworm technology.
abstract: Bombyx mori (silkworm) silk proteins are being utilized as unique biomaterials for medical applications. Chemical modification or post-conjugation of bioactive ligands expand the applicability of silk proteins; however, the processes are elaborate and costly. In this study, we used transgenic silkworm technology to develop single-chain variable fragment (scFv)-conjugated silk fibroin. The cocoons of the transgenic silkworm contain fibroin L-chain linked with scFv as a fusion protein. After dissolving the cocoons in lithium bromide, the silk solution was dialyzed, concentrated, freeze-dried, and crushed into powder. Immunoprecipitation analyses demonstrate that the scFv domain retains its specific binding activity to the target molecule after multiple processing steps. These results strongly suggest the promise of scFv-conjugated silk fibroin as an alternative affinity reagent, which can be manufactured using transgenic silkworm technology at lower cost than traditional affinity carriers.
authors.affiliation: Animal Immune and Cell Biology Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Sato
authors.forename: Katsura
authors.initials: K
authors.lastname: Kojima
authors.forename: Chisato
authors.initials: C
authors.lastname: Sakuma
authors.forename: Maria
authors.initials: M
authors.lastname: Murakami
authors.forename: Eriko
authors.initials: E
authors.lastname: Aratani
authors.forename: Takato
authors.initials: T
authors.lastname: Takenouchi
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Tamada
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kitani
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000378:metabolism; D001965:Bromides / Q000737:chemistry; D005349:Fibroins / Q000096:biosynthesis* / Q000737:chemistry; D018020:Lithium Compounds / Q000737:chemistry; D011208:Powders / Q000737:chemistry; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000737:chemistry
chemical_list: D001965:Bromides; D018020:Lithium Compounds; D011208:Powders; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; C040949:lithium bromide; D005349:Fibroins