Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis

[EN]The production of artificial anti-CB1 antibodies in nanoparticle format is described using the solid-phase imprinting approach. Instead of whole protein imprinting, a linear C-terminus sequence of the receptor comprising 15 amino acids (458-KVTMSVSTDTSAEAL-472) has been used as template, in acco...

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Autores: Gómez Caballero, Alberto, Elejaga Jimeno, Ainhoa, García del Caño, Gontzal, Unceta Zaballa, Nora, Guerreiro, Antonio, Saumell Esnaola, Miquel, Sallés Alvira, Joan, Goicolea Altuna, María Aranzazu, Barrio Díez-Caballero, Ramón José
Tipo de recurso: artículo
Fecha de publicación:2021
País:España
Institución:Universidad del País Vasco
Repositorio:Addi. Archivo Digital para la Docencia y la Investigación
OAI Identifier:oai:addi.ehu.eus:10810/54158
Acceso en línea:http://hdl.handle.net/10810/54158
Access Level:acceso abierto
Palabra clave:artificial antibody
epitope imprinting
GPCR
CB1 receptor
molecularly imprinted nanoparticles
bioanalysis
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spelling Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysisGómez Caballero, AlbertoElejaga Jimeno, AinhoaGarcía del Caño, GontzalUnceta Zaballa, NoraGuerreiro, AntonioSaumell Esnaola, MiquelSallés Alvira, JoanGoicolea Altuna, María AranzazuBarrio Díez-Caballero, Ramón Joséartificial antibodyepitope imprintingGPCRCB1 receptormolecularly imprinted nanoparticlesbioanalysis[EN]The production of artificial anti-CB1 antibodies in nanoparticle format is described using the solid-phase imprinting approach. Instead of whole protein imprinting, a linear C-terminus sequence of the receptor comprising 15 amino acids (458-KVTMSVSTDTSAEAL-472) has been used as template, in accordance with the epitope imprinting approach. This sequence is located intracellularly, and it is involved in coupling to G(i/o) proteins, being responsible for CB1 receptor desensitisation and internalisation. Developed molecularly imprinted materials were found to be in the nanometre scale, with a particle size of 126.4 +/- 10.5 nm at pH 3 (25 oC) and spherical shape. It was also observed that the size was sensible to temperature changes being reduced to 106.3 +/- 15.2 nm at 35 degrees C. Lower critical solution temperature of this polymer was found to be approximate to 33.4 degrees C. The affinity and selectivity of the artificial antibody were assessed through dot blot and Western blot experiments. For the latter, recombinant fusion proteins GST-CB1(414-472) and GST-CB1(414-442) were produced to work respectively as target and negative control proteins. The control protein did not carry the target epitope for being devoid of last 30 amino acids at the C-terminus. The results demonstrated that the anti-CB1 material recognised selectively the target protein, thanks to the presence of the 15-amino acid sequence selected as epitope, which revealed that binding occurred at the C-terminus of the receptor itself. The methodology presented may pave the way for the development of novel imprinted nanomaterials for other proteins included in the superfamily of the G-protein-coupled receptors (GPCR).Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. Funding for this research was provided by the Spanish Ministry of Science, Innovation and Universities (project CTQ2017-85686-R) and by the Basque Government (Research Groups of the Basque University System, Project No IT 1186-19.Springer202120212021info:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10810/54158reponame:Addi. Archivo Digital para la Docencia y la Investigacióninstname:Universidad del País VascoInglésinfo:eu-repo/grantAgreement/MINECO/CTQ2017-85686-R/https://link.springer.com/article/10.1007%2Fs00604-021-05029-zinfo:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/3.0/es/© The Author(s) 2021. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.Atribución 3.0 Españaoai:addi.ehu.eus:10810/541582026-06-18T09:23:17Z
dc.title.none.fl_str_mv Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis
title Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis
spellingShingle Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis
Gómez Caballero, Alberto
artificial antibody
epitope imprinting
GPCR
CB1 receptor
molecularly imprinted nanoparticles
bioanalysis
title_short Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis
title_full Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis
title_fullStr Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis
title_full_unstemmed Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis
title_sort Solid-phase synthesis of imprinted nanoparticles as artificial antibodies against the C-terminus of the cannabinoid CB1 receptor: exploring a viable alternative for bioanalysis
dc.creator.none.fl_str_mv Gómez Caballero, Alberto
Elejaga Jimeno, Ainhoa
García del Caño, Gontzal
Unceta Zaballa, Nora
Guerreiro, Antonio
Saumell Esnaola, Miquel
Sallés Alvira, Joan
Goicolea Altuna, María Aranzazu
Barrio Díez-Caballero, Ramón José
author Gómez Caballero, Alberto
author_facet Gómez Caballero, Alberto
Elejaga Jimeno, Ainhoa
García del Caño, Gontzal
Unceta Zaballa, Nora
Guerreiro, Antonio
Saumell Esnaola, Miquel
Sallés Alvira, Joan
Goicolea Altuna, María Aranzazu
Barrio Díez-Caballero, Ramón José
author_role author
author2 Elejaga Jimeno, Ainhoa
García del Caño, Gontzal
Unceta Zaballa, Nora
Guerreiro, Antonio
Saumell Esnaola, Miquel
Sallés Alvira, Joan
Goicolea Altuna, María Aranzazu
Barrio Díez-Caballero, Ramón José
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv artificial antibody
epitope imprinting
GPCR
CB1 receptor
molecularly imprinted nanoparticles
bioanalysis
topic artificial antibody
epitope imprinting
GPCR
CB1 receptor
molecularly imprinted nanoparticles
bioanalysis
description [EN]The production of artificial anti-CB1 antibodies in nanoparticle format is described using the solid-phase imprinting approach. Instead of whole protein imprinting, a linear C-terminus sequence of the receptor comprising 15 amino acids (458-KVTMSVSTDTSAEAL-472) has been used as template, in accordance with the epitope imprinting approach. This sequence is located intracellularly, and it is involved in coupling to G(i/o) proteins, being responsible for CB1 receptor desensitisation and internalisation. Developed molecularly imprinted materials were found to be in the nanometre scale, with a particle size of 126.4 +/- 10.5 nm at pH 3 (25 oC) and spherical shape. It was also observed that the size was sensible to temperature changes being reduced to 106.3 +/- 15.2 nm at 35 degrees C. Lower critical solution temperature of this polymer was found to be approximate to 33.4 degrees C. The affinity and selectivity of the artificial antibody were assessed through dot blot and Western blot experiments. For the latter, recombinant fusion proteins GST-CB1(414-472) and GST-CB1(414-442) were produced to work respectively as target and negative control proteins. The control protein did not carry the target epitope for being devoid of last 30 amino acids at the C-terminus. The results demonstrated that the anti-CB1 material recognised selectively the target protein, thanks to the presence of the 15-amino acid sequence selected as epitope, which revealed that binding occurred at the C-terminus of the receptor itself. The methodology presented may pave the way for the development of novel imprinted nanomaterials for other proteins included in the superfamily of the G-protein-coupled receptors (GPCR).
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10810/54158
url http://hdl.handle.net/10810/54158
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/MINECO/CTQ2017-85686-R/
https://link.springer.com/article/10.1007%2Fs00604-021-05029-z
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/3.0/es/
Atribución 3.0 España
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/3.0/es/
Atribución 3.0 España
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv reponame:Addi. Archivo Digital para la Docencia y la Investigación
instname:Universidad del País Vasco
instname_str Universidad del País Vasco
reponame_str Addi. Archivo Digital para la Docencia y la Investigación
collection Addi. Archivo Digital para la Docencia y la Investigación
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300719

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