{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,21]],"date-time":"2026-03-21T18:02:25Z","timestamp":1774116145654,"version":"3.50.1"},"reference-count":57,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2017,4,19]],"date-time":"2017-04-19T00:00:00Z","timestamp":1492560000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000288","name":"Royal Society","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100000288","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004440","name":"Wellcome Trust","doi-asserted-by":"publisher","award":["WT093092MA"],"award-info":[{"award-number":["WT093092MA"]}],"id":[{"id":"10.13039\/100004440","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010663","name":"H2020 European Research Council","doi-asserted-by":"publisher","award":["ERC\u20102010\u2010AdG_20100317"],"award-info":[{"award-number":["ERC\u20102010\u2010AdG_20100317"]}],"id":[{"id":"10.13039\/100010663","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Plant Biotechnology Journal"],"published-print":{"date-parts":[[2017,10]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>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 <jats:italic>in planta<\/jats:italic> for targeted delivery across the blood\u2013brain barrier (<jats:styled-content style=\"fixed-case\">BBB<\/jats:styled-content>). Rabies virus (<jats:styled-content style=\"fixed-case\">RABV<\/jats:styled-content>) 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 <jats:styled-content style=\"fixed-case\">RABV<\/jats:styled-content> glycoprotein peptide sequence to assist delivery of a rabies neutralizing single\u2010chain antibody (ScFv) across an <jats:italic>in cellulo<\/jats:italic> model of human <jats:styled-content style=\"fixed-case\">BBB<\/jats:styled-content>. The 29 amino acid rabies virus peptide (<jats:styled-content style=\"fixed-case\">RVG<\/jats:styled-content>) recognizes the nicotinic acetylcholine receptor (<jats:styled-content style=\"fixed-case\">nA<\/jats:styled-content>chR) at neuromuscular junctions and the <jats:styled-content style=\"fixed-case\">BBB<\/jats:styled-content>. ScFv and ScFv\u2010<jats:styled-content style=\"fixed-case\">RVG<\/jats:styled-content> fusion proteins were produced in <jats:italic>Nicotiana benthamiana<\/jats:italic> by transient expression. Both molecules were successfully expressed and purified, but the ScFv expression level was significantly higher than that of ScFv\u2010<jats:styled-content style=\"fixed-case\">RVG<\/jats:styled-content> fusion. Both ScFv and ScFv\u2010<jats:styled-content style=\"fixed-case\">RVG<\/jats:styled-content> fusion molecules had potent neutralization activity against <jats:styled-content style=\"fixed-case\">RABV<\/jats:styled-content><jats:italic>in cellulo<\/jats:italic>. The ScFv\u2010<jats:styled-content style=\"fixed-case\">RVG<\/jats:styled-content> fusion demonstrated increased binding to <jats:styled-content style=\"fixed-case\">nA<\/jats:styled-content>chR and entry into neuronal cells, compared to ScFv alone. Additionally, a human brain endothelial cell line <jats:styled-content style=\"fixed-case\">BBB<\/jats:styled-content> model was used to demonstrate that plant\u2010produced ScFv\u2010<jats:styled-content style=\"fixed-case\">RVG<jats:sup>P<\/jats:sup><\/jats:styled-content> fusion could translocate across the cells. This study indicates that the plant\u2010produced ScFv\u2010<jats:styled-content style=\"fixed-case\">RVG<jats:sup>P<\/jats:sup><\/jats:styled-content> fusion protein was able to cross the <jats:italic>in cellulo<\/jats:italic><jats:styled-content style=\"fixed-case\">BBB<\/jats:styled-content> and neutralize RABV.<\/jats:p>","DOI":"10.1111\/pbi.12719","type":"journal-article","created":{"date-parts":[[2017,3,8]],"date-time":"2017-03-08T18:36:43Z","timestamp":1488998203000},"page":"1331-1339","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":21,"title":["Enhanced transport of plant\u2010produced rabies single\u2010chain antibody\u2010<scp>RVG<\/scp> peptide fusion protein across an <i>in cellulo<\/i> blood\u2013brain barrier device"],"prefix":"10.1111","volume":"15","author":[{"given":"Waranyoo","family":"Phoolcharoen","sequence":"first","affiliation":[{"name":"Institute for Infection and Immunity St. George's Hospital Medical School University of London London UK"},{"name":"Pharmacognosy and Pharmaceutical Botany Faculty of Pharmaceutical Sciences Chulalongkorn University Bangkok Thailand"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christophe","family":"Prehaud","sequence":"additional","affiliation":[{"name":"Unit\u00e9 de Neuroimmunologie Virale D\u00e9partement de Virologie Institut Pasteur Paris France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Craig J.","family":"van Dolleweerd","sequence":"additional","affiliation":[{"name":"Institute for Infection and Immunity St. George's Hospital Medical School University of London London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Leonard","family":"Both","sequence":"additional","affiliation":[{"name":"Institute for Infection and Immunity St. George's Hospital Medical School University of London London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anaelle","family":"da Costa","sequence":"additional","affiliation":[{"name":"Unit\u00e9 de Neuroimmunologie Virale D\u00e9partement de Virologie Institut Pasteur Paris France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Monique","family":"Lafon","sequence":"additional","affiliation":[{"name":"Unit\u00e9 de Neuroimmunologie Virale D\u00e9partement de Virologie Institut Pasteur Paris France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Julian K\u2010C.","family":"Ma","sequence":"additional","affiliation":[{"name":"Institute for Infection and Immunity St. George's Hospital Medical School University of London London UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,4,19]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11095-014-1519-8"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1012098005098"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.3109\/02652049809006836"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.3140379"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2010.01.011"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1096\/fj.12-219964"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/0161-5890(88)90125-3"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/0168-1702(85)90014-0"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13365-011-0041-6"},{"key":"e_1_2_6_11_1","first-page":"221","article-title":"Synthetic peptides of neurotoxins and rabies virus glycoprotein behave as antagonists in a functional assay for the acetylcholine receptor","volume":"2","author":"Donnelly\u2010Roberts D.L.","year":"1989","journal-title":"Pept. Res."},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1186\/2045-8118-10-33"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(13)62707-5"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.11.4771"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11095-012-0667-y"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.73.7.2424"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1018983904537"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1002\/biot.201300162"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1080\/21688370.2016.1142493"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1474-4422(02)00041-8"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2011.03.047"},{"key":"e_1_2_6_22_1","first-page":"71","article-title":"Pathogenetic mechanisms of septic states in war injuries","volume":"26","author":"Jekic M.","year":"1979","journal-title":"Acta Chir. Iugosl."},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2012.09.047"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2013.273"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature05901"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2016.115"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmr.300030205"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00109a020"},{"key":"e_1_2_6_29_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.7053569"},{"key":"e_1_2_6_30_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.6494916"},{"key":"e_1_2_6_31_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.340020406"},{"key":"e_1_2_6_32_1","first-page":"57","article-title":"Synthetic peptides in the study of the interaction of rabies virus and the acetylcholine receptor","volume":"44","author":"Lentz T.L.","year":"1988","journal-title":"Adv. Biochem. Psychopharmacol."},{"key":"e_1_2_6_33_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-4598(200005)23:5<720::AID-MUS9>3.0.CO;2-5"},{"key":"e_1_2_6_34_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0066084"},{"key":"e_1_2_6_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biomaterials.2009.02.051"},{"key":"e_1_2_6_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/0092-1157(75)90061-X"},{"key":"e_1_2_6_37_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.lfs.2014.08.006"},{"key":"e_1_2_6_38_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejpb.2010.11.015"},{"key":"e_1_2_6_39_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(20)81864-X"},{"key":"e_1_2_6_40_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2005.06.027"},{"key":"e_1_2_6_41_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pntd.0000542"},{"key":"e_1_2_6_42_1","doi-asserted-by":"publisher","DOI":"10.3109\/10611860903548354"},{"key":"e_1_2_6_43_1","doi-asserted-by":"publisher","DOI":"10.1517\/17425247.2014.952627"},{"key":"e_1_2_6_44_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1471-4159.1985.tb07167.x"},{"key":"e_1_2_6_45_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11103-013-0036-1"},{"key":"e_1_2_6_46_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12033-015-9880-3"},{"key":"e_1_2_6_47_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pep.2009.10.002"},{"key":"e_1_2_6_48_1","doi-asserted-by":"publisher","DOI":"10.1002\/jmr.300020202"},{"key":"e_1_2_6_49_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1467-7652.2009.00434.x"},{"key":"e_1_2_6_50_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.285.5433.1569"},{"key":"e_1_2_6_51_1","doi-asserted-by":"publisher","DOI":"10.1111\/pbi.12604"},{"key":"e_1_2_6_52_1","doi-asserted-by":"publisher","DOI":"10.1016\/B978-0-12-387040-7.00013-5"},{"key":"e_1_2_6_53_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcp.1041320115"},{"key":"e_1_2_6_54_1","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.71.10.7372-7380.1997"},{"key":"e_1_2_6_55_1","doi-asserted-by":"publisher","DOI":"10.1002\/j.1460-2075.1983.tb01434.x"},{"key":"e_1_2_6_56_1","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cis384"},{"key":"e_1_2_6_57_1","doi-asserted-by":"publisher","DOI":"10.3109\/1061186X.2010.523788"},{"key":"e_1_2_6_58_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0016519"}],"container-title":["Plant Biotechnology Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fpbi.12719","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/pbi.12719","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,11,20]],"date-time":"2024-11-20T12:35:02Z","timestamp":1732106102000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/pbi.12719"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,4,19]]},"references-count":57,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2017,10]]}},"alternative-id":["10.1111\/pbi.12719"],"URL":"https:\/\/doi.org\/10.1111\/pbi.12719","archive":["Portico"],"relation":{},"ISSN":["1467-7644","1467-7652"],"issn-type":[{"value":"1467-7644","type":"print"},{"value":"1467-7652","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,4,19]]},"assertion":[{"value":"2016-06-17","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2017-03-01","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2017-04-19","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}