Directed Functionalization of Recombinant Spider Silk Nonwoven Membranes with Antibodies Using Non-Canonical Amino Acids

Natural spider silk fibers are recognized for their outstanding mechanical properties. The production of underlying recombinant spider silk proteins in scalable quantities unlocks their potential for technical and biomedical applications. The recombinant spider silk technology further enables the introduction of new functions via genetic encoding. In the present study, the non-canonical amino acid L-azidohomoalanine is incorporated into the engineered Araneus diadematus fibroin 4, eADF4(C16), at positions explicitly defined by methionine codons in an N-terminal peptide tag. The azido-functionalized eADF4(C16) is processed into particles, films, or electrospun into nanofibers. As functional entities, high molecular weight molecules, such as antibodies, are modified with bio-orthogonal groups suitable for strain-promoted cycloaddition to the exposed azido-groups on the spider silk supports. The antibody-activated spider silk nonwoven meshes exemplarily show their applicability as bio-capturing membranes.