Production of scFv-conjugated affinity silk film and its application to a novel enzyme-linked immunosorbent assay.

[wiskott-aldrich syndrome]

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 .