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. 2023 Aug 20:669:77-84.
doi: 10.1016/j.bbrc.2023.05.093. Epub 2023 May 24.

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

Linnea Charlotta Hjelm  1 , My Hedhammar  2 , John Löfblom  3
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In vitro Blood-Brain barrier model based on recombinant spider silk protein nanomembranes for evaluation of transcytosis capability of biomolecules

Linnea Charlotta Hjelm et al. Biochem Biophys Res Commun. .
Free article

Abstract

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 engineered shuttle proteins facilitates ranking and the selection of promising candidates during development. 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.

Keywords: Blood–brain barrier; Receptor-mediated transcytosis; Recombinant spider silk; Transferrin Receptor-1; Transwell assay.

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Conflict of interest statement

Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests. My Hedhammar reports a relationship with Spiber Technologies AB that includes: equity or stocks.

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