Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening
| Autor(a) principal: | |
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| Data de Publicação: | 2021 |
| Outros Autores: | , , , , , , , , , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| Texto Completo: | http://hdl.handle.net/10451/57424 |
Resumo: | © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
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Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screeningBrain targeting antibodiesBlood-brain barrierin vivo phage displaySingle-domain antibodiesDrug delivery© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).A major bottleneck in the successful development of central nervous system (CNS) drugs is the discovery and design of molecules that can cross the blood-brain barrier (BBB). Nano-delivery strategies are a promising approach that take advantage of natural portals of entry into the brain such as monoclonal antibodies (mAbs) targeting endogenous BBB receptors. However, the main selected mAbs rely on targeting broadly expressed receptors, such as the transferrin and insulin receptors, and in selection processes that do not fully mimic the native receptor conformation, leading to mistargeting and a low fraction of the administered dose effectively reaching the brain. Thus, there is an urgent need to identify new BBB receptors and explore novel antibody selection approaches that can allow a more selective delivery into the brain. Considering that in vitro models fail to completely mimic brain structure complexity, we explored an in vivo cell immunization approach to construct a rabbit derived single-domain antibody (sdAb) library towards BBB endothelial cell receptors. The sdAb antibody library was used in an in vivo phage display screening as a functional selection of novel BBB targeting antibodies. Following three rounds of selections, next generation sequencing analysis, in vitro brain endothelial barrier (BEB) model screenings and in vivo biodistribution studies, five potential sdAbs were identified, three of which reaching >0.6% ID/g in the brain. To validate the brain drug delivery proof-of-concept, the most promising sdAb, namely RG3, was conjugated at the surface of liposomes encapsulated with a model drug, the pan-histone deacetylase inhibitor panobinostat (PAN). The translocation efficiency and activity of the conjugate liposome was determined in a dual functional in vitro BEB-glioblastoma model. The RG3 conjugated PAN liposomes enabled an efficient BEB translocation and presented a potent antitumoral activity against LN229 glioblastoma cells without influencing BEB integrity. In conclusion, our in vivo screening approach allowed the selection of highly specific nano-antibody scaffolds with promising properties for brain targeting and drug delivery.This research was funded by: the Portuguese Funding Agency, Fundacão para a Ciência e a Tecnologia, FCT IP: IF/01010/2013 and PTDC/BBB-BIO/0508/2014 to FAS, SFRH/BPD/100522/2014 and DL57/2016/CP1438/CT0002 to SIA, PD/BD/128281/2017, PTDC/BIA-BQM/5027/2020 and Dl 57/2016/CP1451/CT0023 to MC and VN, UIDB/04138/2020 and UIDP/04138/2020 to MMG, UID/Multi/04349/2020 and PTDC/QUI-NUC/30147/2017 to LG and JDGC, UID/NEU/04539/2019, UIDB/04539/2020 and ViraVector (CENTRO-01-0145-FEDER-022095) to RJN and LPA. Gilead Génese 2019 has provided support through project PGG/050/2019 to JNRD. CIISA has provided support through Project UIDB/00276/2020, funded by FCT. FAS and JG dedicate this study to the memory of Professor Carlos F. Barbas 3rd, an outstanding mentor and friend.MDPIRepositório da Universidade de LisboaAguiar, Sandra IDias, Joana N. R.André, AnaSilva, MartaMartins, DianaCarrapiço, BelmiraCastanho, Miguel A. R. B.Carrico, Joao AndreCavaco, MarcoGaspar, Maria ManuelaNobre, Rui JorgePereira de Almeida, LuísOliveira, SoraiaGano, LurdesCorreia, João D. G.Carlos F. Barbas, IIIGonçalves, João RafaelNeves, VeraAires da Silva, Frederico2023-05-15T13:30:04Z20212021-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10451/57424engPharmaceutics 2021, 13, 159810.3390/pharmaceutics131015981999-4923info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-08T17:05:40Zoai:repositorio.ul.pt:10451/57424Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T22:07:50.531707Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
| dc.title.none.fl_str_mv |
Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening |
| title |
Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening |
| spellingShingle |
Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening Aguiar, Sandra I Brain targeting antibodies Blood-brain barrier in vivo phage display Single-domain antibodies Drug delivery |
| title_short |
Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening |
| title_full |
Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening |
| title_fullStr |
Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening |
| title_full_unstemmed |
Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening |
| title_sort |
Highly specific blood-brain barrier transmigrating single-domain antibodies selected by an in vivo phage display screening |
| author |
Aguiar, Sandra I |
| author_facet |
Aguiar, Sandra I Dias, Joana N. R. André, Ana Silva, Marta Martins, Diana Carrapiço, Belmira Castanho, Miguel A. R. B. Carrico, Joao Andre Cavaco, Marco Gaspar, Maria Manuela Nobre, Rui Jorge Pereira de Almeida, Luís Oliveira, Soraia Gano, Lurdes Correia, João D. G. Carlos F. Barbas, III Gonçalves, João Rafael Neves, Vera Aires da Silva, Frederico |
| author_role |
author |
| author2 |
Dias, Joana N. R. André, Ana Silva, Marta Martins, Diana Carrapiço, Belmira Castanho, Miguel A. R. B. Carrico, Joao Andre Cavaco, Marco Gaspar, Maria Manuela Nobre, Rui Jorge Pereira de Almeida, Luís Oliveira, Soraia Gano, Lurdes Correia, João D. G. Carlos F. Barbas, III Gonçalves, João Rafael Neves, Vera Aires da Silva, Frederico |
| author2_role |
author author author author author author author author author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
Repositório da Universidade de Lisboa |
| dc.contributor.author.fl_str_mv |
Aguiar, Sandra I Dias, Joana N. R. André, Ana Silva, Marta Martins, Diana Carrapiço, Belmira Castanho, Miguel A. R. B. Carrico, Joao Andre Cavaco, Marco Gaspar, Maria Manuela Nobre, Rui Jorge Pereira de Almeida, Luís Oliveira, Soraia Gano, Lurdes Correia, João D. G. Carlos F. Barbas, III Gonçalves, João Rafael Neves, Vera Aires da Silva, Frederico |
| dc.subject.por.fl_str_mv |
Brain targeting antibodies Blood-brain barrier in vivo phage display Single-domain antibodies Drug delivery |
| topic |
Brain targeting antibodies Blood-brain barrier in vivo phage display Single-domain antibodies Drug delivery |
| description |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
| publishDate |
2021 |
| dc.date.none.fl_str_mv |
2021 2021-01-01T00:00:00Z 2023-05-15T13:30:04Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
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http://hdl.handle.net/10451/57424 |
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http://hdl.handle.net/10451/57424 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
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Pharmaceutics 2021, 13, 1598 10.3390/pharmaceutics13101598 1999-4923 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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MDPI |
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MDPI |
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