Open AccessEnhanced transport of plant‐produced rabies single‐chain antibody‐ RVG peptide fusion protein across an in cellulo blood–brain barrier device
Author(s) -
Phoolcharoen Waranyoo,
Prehaud Christophe,
Dolleweerd Craig J.,
Both Leonard,
Costa Anaelle,
Lafon Monique,
Ma Julian KC.
Publication year - 2017
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/pbi.12719
Subject(s) - biology , fusion protein , antibody , peptide , single chain , microbiology and biotechnology , rabies , fusion , virology , computational biology , biochemistry , immunology , gene , recombinant dna , linguistics , philosophy
Summary The biomedical applications of antibody engineering are developing rapidly and have been expanded to plant expression platforms. In this study, we have generated a novel antibody molecule in planta for targeted delivery across the blood–brain barrier ( BBB ). Rabies virus ( RABV ) is a neurotropic virus for which there is no effective treatment after entry into the central nervous system. This study investigated the use of a RABV glycoprotein peptide sequence to assist delivery of a rabies neutralizing single‐chain antibody (ScFv) across an in cellulo model of human BBB . The 29 amino acid rabies virus peptide ( RVG ) recognizes the nicotinic acetylcholine receptor ( nA chR) at neuromuscular junctions and the BBB . ScFv and ScFv‐ RVG fusion proteins were produced in Nicotiana benthamiana by transient expression. Both molecules were successfully expressed and purified, but the ScFv expression level was significantly higher than that of ScFv‐ RVG fusion. Both ScFv and ScFv‐ RVG fusion molecules had potent neutralization activity against RABV in cellulo . The ScFv‐ RVG fusion demonstrated increased binding to nA chR and entry into neuronal cells, compared to ScFv alone. Additionally, a human brain endothelial cell line BBB model was used to demonstrate that plant‐produced ScFv‐ RVG P fusion could translocate across the cells. This study indicates that the plant‐produced ScFv‐ RVG P fusion protein was able to cross the in cellulo BBB and neutralize RABV.