In vitro Blood-Brain barrier model based on recombinant spider silk protein nanomembranes for evaluation of transcytosis capability of biomolecules

The blood-brain barrier (BBB) limits the uptake of central nervous system (CNS)-targeting drugs into the brain. Engineering molecular shuttles for active transportation across the barrier has thus potential for improving the efficacy of such drugs. In vitro assessment of potential transcytosis capability for engi-neered shuttle proteins facilitates ranking and the selection of promising candidates during develop-ment. Herein, the development of an assay based on brain endothelial cells cultured on permeable recombinant silk nanomembranes for screening of transcytosis capability of biomolecules is described. The silk nanomembranes supported growth of brain endothelial cells to form confluent monolayers with relevant cell morphology, and induced expression of tight-junction proteins. Evaluation of the assay using an established BBB shuttle antibody showed transcytosis over the membranes with an apparent permeability that significantly differed from the isotype control antibody.& COPY; 2023 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Automated summary

The development of an assay based on brain endothelial cells cultured on permeable recombinant silk nanomembranes for screening the transcytosis capability of biomolecules is described. Evaluation of the assay using an established BBB shuttle antibody showed significant transcytosis over the membranes.