Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)

The development of a new generation of non-antibody protein drug delivery systems requires site-directed conjugation strategies to produce homogeneous, reproducible and scalable nanomedicines. For that, the genetic addition of cysteine residues into solvent-exposed positions allows the thiol-mediate...

Descripción completa

Detalles Bibliográficos
Autores: Rueda A., Mendoza J.I., Alba-Castellon L., Parladé E., Voltà-Durán E., Paez D., Aviño A., Eritja R., Vázquez E., Villaverde A., Mangues R., Unzueta U.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2023
País:España
Institución:Institut d’Investigació Biomèdica Sant Pau (IIB Sant Pau)
Repositorio:r-IIB SANT PAU. Repositorio Institucional de Producción Científica del Instituto de Investigación Biomédica Sant Pau
OAI Identifier:oai:iibsantpau.fundanetsuite.com:p17216
Acceso en línea:https://iibsantpau.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=17216
https://www.scopus.com/inward/record.uri?eid=2-s2.0-85172098368&doi=10.1007%2fs40843-023-2571-6&partnerID=40&md5=745cd7379ee2ea05cc0ec7cff70ee5fd
Access Level:acceso abierto
Palabra clave:nanomedicine
protein carriers
disulfide-bonds
THIOCAP
nanoconjugates
cysteine coupling
id ES_aa5da89a40ef54655aebb47b0f77d0a1
oai_identifier_str oai:iibsantpau.fundanetsuite.com:p17216
network_acronym_str ES
network_name_str España
repository_id_str
spelling Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)Rueda A.Mendoza J.I.Alba-Castellon L.Parladé E.Voltà-Durán E.Paez D.Aviño A.Eritja R.Vázquez E.Villaverde A.Mangues R.Unzueta U.nanomedicineprotein carriersdisulfide-bondsTHIOCAPnanoconjugatescysteine couplingThe development of a new generation of non-antibody protein drug delivery systems requires site-directed conjugation strategies to produce homogeneous, reproducible and scalable nanomedicines. For that, the genetic addition of cysteine residues into solvent-exposed positions allows the thiol-mediated cysteine coupling of therapeutic drugs into protein-based nanocarriers. However, the high reactivity of unpaired cysteine residues usually reduces protein stability, consequently imposing the use of more methodologically demanding purification procedures. This is especially relevant for disulfide-containing nanocarriers, as previously observed in THIOMABs. Moreover, although many protein scaffolds and targeting ligands are also rich in disulfide bridges, the use of these methodologies over emerging non-antibody carrier proteins has been completely neglected. Here, we report the development of a simple and straightforward procedure for a one-step production and site-directed cysteine conjugation of disulfide-containing non-antibody thiolated carrier proteins (THIOCAPs). This method is validated in a fluorescent C-X-C chemokine receptor 4 (CXCR4)-targeted multivalent nano-carrier containing two intramolecular disulfide bridges and one reactive cysteine residue strategically placed into a solvent-exposed position (THIO-T22-GFP-H6) for drug conjugation and in a humanized alternative intended for clinical applications (T22-HSNBT-H6). Thus, we produce very stable, homogeneous and fully functional antitumoral nanoconjugates (THIO-T22-GFP-H6-MMAE and T22-HSNBT-H6-MMAE) that selectively eliminate target cancer cells via CXCR4-receptor. Altogether, the developed methodology appears as a powerful tool for the rational engineering of emerging non-antibody, cell-targeted protein nanocarriers that contain disulfide bridges together with a solvent-exposed reactive cysteine (THIOCAP). This should pave the way for the development of a new generation of stable, homogeneous and efficient nanomedicines.SCIENCE PRESS2023info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://iibsantpau.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=17216https://www.scopus.com/inward/record.uri?eid=2-s2.0-85172098368&doi=10.1007%2fs40843-023-2571-6&partnerID=40&md5=745cd7379ee2ea05cc0ec7cff70ee5fdScience China-MaterialsISSN: 20958226ISSNe: 21994501reponame:r-IIB SANT PAU. Repositorio Institucional de Producción Científica del Instituto de Investigación Biomédica Sant Pauinstname:Institut d’Investigació Biomèdica Sant Pau (IIB Sant Pau)Inglésinfo:eu-repo/semantics/openAccessoai:iibsantpau.fundanetsuite.com:p172162026-06-14T12:41:47Z
dc.title.none.fl_str_mv Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)
title Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)
spellingShingle Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)
Rueda A.
nanomedicine
protein carriers
disulfide-bonds
THIOCAP
nanoconjugates
cysteine coupling
title_short Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)
title_full Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)
title_fullStr Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)
title_full_unstemmed Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)
title_sort Site-directed cysteine coupling of disulfide-containing non-antibody carrier proteins (THIOCAPs)
dc.creator.none.fl_str_mv Rueda A.
Mendoza J.I.
Alba-Castellon L.
Parladé E.
Voltà-Durán E.
Paez D.
Aviño A.
Eritja R.
Vázquez E.
Villaverde A.
Mangues R.
Unzueta U.
author Rueda A.
author_facet Rueda A.
Mendoza J.I.
Alba-Castellon L.
Parladé E.
Voltà-Durán E.
Paez D.
Aviño A.
Eritja R.
Vázquez E.
Villaverde A.
Mangues R.
Unzueta U.
author_role author
author2 Mendoza J.I.
Alba-Castellon L.
Parladé E.
Voltà-Durán E.
Paez D.
Aviño A.
Eritja R.
Vázquez E.
Villaverde A.
Mangues R.
Unzueta U.
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv nanomedicine
protein carriers
disulfide-bonds
THIOCAP
nanoconjugates
cysteine coupling
topic nanomedicine
protein carriers
disulfide-bonds
THIOCAP
nanoconjugates
cysteine coupling
description The development of a new generation of non-antibody protein drug delivery systems requires site-directed conjugation strategies to produce homogeneous, reproducible and scalable nanomedicines. For that, the genetic addition of cysteine residues into solvent-exposed positions allows the thiol-mediated cysteine coupling of therapeutic drugs into protein-based nanocarriers. However, the high reactivity of unpaired cysteine residues usually reduces protein stability, consequently imposing the use of more methodologically demanding purification procedures. This is especially relevant for disulfide-containing nanocarriers, as previously observed in THIOMABs. Moreover, although many protein scaffolds and targeting ligands are also rich in disulfide bridges, the use of these methodologies over emerging non-antibody carrier proteins has been completely neglected. Here, we report the development of a simple and straightforward procedure for a one-step production and site-directed cysteine conjugation of disulfide-containing non-antibody thiolated carrier proteins (THIOCAPs). This method is validated in a fluorescent C-X-C chemokine receptor 4 (CXCR4)-targeted multivalent nano-carrier containing two intramolecular disulfide bridges and one reactive cysteine residue strategically placed into a solvent-exposed position (THIO-T22-GFP-H6) for drug conjugation and in a humanized alternative intended for clinical applications (T22-HSNBT-H6). Thus, we produce very stable, homogeneous and fully functional antitumoral nanoconjugates (THIO-T22-GFP-H6-MMAE and T22-HSNBT-H6-MMAE) that selectively eliminate target cancer cells via CXCR4-receptor. Altogether, the developed methodology appears as a powerful tool for the rational engineering of emerging non-antibody, cell-targeted protein nanocarriers that contain disulfide bridges together with a solvent-exposed reactive cysteine (THIOCAP). This should pave the way for the development of a new generation of stable, homogeneous and efficient nanomedicines.
publishDate 2023
dc.date.none.fl_str_mv 2023
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://iibsantpau.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=17216
https://www.scopus.com/inward/record.uri?eid=2-s2.0-85172098368&doi=10.1007%2fs40843-023-2571-6&partnerID=40&md5=745cd7379ee2ea05cc0ec7cff70ee5fd
url https://iibsantpau.fundanetsuite.com/Publicaciones/ProdCientif/PublicacionFrw.aspx?id=17216
https://www.scopus.com/inward/record.uri?eid=2-s2.0-85172098368&doi=10.1007%2fs40843-023-2571-6&partnerID=40&md5=745cd7379ee2ea05cc0ec7cff70ee5fd
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv SCIENCE PRESS
publisher.none.fl_str_mv SCIENCE PRESS
dc.source.none.fl_str_mv Science China-Materials
ISSN: 20958226
ISSNe: 21994501
reponame:r-IIB SANT PAU. Repositorio Institucional de Producción Científica del Instituto de Investigación Biomédica Sant Pau
instname:Institut d’Investigació Biomèdica Sant Pau (IIB Sant Pau)
instname_str Institut d’Investigació Biomèdica Sant Pau (IIB Sant Pau)
reponame_str r-IIB SANT PAU. Repositorio Institucional de Producción Científica del Instituto de Investigación Biomédica Sant Pau
collection r-IIB SANT PAU. Repositorio Institucional de Producción Científica del Instituto de Investigación Biomédica Sant Pau
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869416157051617280
score 15.198674

Similares en LA Referencia