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Creation and evaluation of a single‐chain antibody tetramer that targets brain endothelial cells
Author(s) -
Zhang Xiaobin,
Wang Xin Xiang,
Shusta Eric V.
Publication year - 2014
Publication title -
aiche journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.958
H-Index - 167
eISSN - 1547-5905
pISSN - 0001-1541
DOI - 10.1002/aic.14348
Subject(s) - biotinylation , internalization , streptavidin , tetramer , blood–brain barrier , chemistry , antibody , microbiology and biotechnology , yeast , endothelial stem cell , cell , biochemistry , biotin , biology , enzyme , immunology , neuroscience , central nervous system , in vitro
Antibodies that target and internalize into blood‐brain barrier (BBB) endothelial cells offer promise as drug delivery agents. Previously, we identified a single‐chain antibody (scFvA) capable of binding to the BBB. In an attempt to improve the binding and internalization properties of scFvA, a biotinylation tag (Avitag) was fused to scFvA and the protein secreted by yeast. The scFvA‐Avitag could be biotinylated by yeast‐displayed BirA enzyme and biotinylated scFvA‐Avitag could be used to create scFv tetramers. Tetramerization of scFvA improved the internalization of scFvA into BBB endothelial cells, and biotinylated scFvA‐Avitag could also be used to target streptavidin‐coated quantum dots for BBB endothelial cell internalization. Perfusing the rat brain with scFvA‐tetramer confirmed that the antigen targeted by scFvA is distributed on the blood side of the BBB, suggesting the potential for downstream application of scFvA in brain‐targeted drug delivery. © 2014 American Institute of Chemical Engineers AIChE J , 60: 1245–1252, 2014