Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form

© 2021 by the authors.

Bibliographic Details
Authors: Artusio, Fiora, Castellví, Albert, Pisano, Roberto, Gavira Gallardo, J. A.
Format: article
Status:Published version
Publication Date:2021
Country:España
Institution:Consejo Superior de Investigaciones Científicas (CSIC)
Repository:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/254808
Online Access:http://hdl.handle.net/10261/254808
Access Level:Open access
Keyword:Protein
Crystallization
Polymorphism
Agarose gel
Nucleation
id ES_fbcfbd956de16b2cca7c2ae87cc6405f
oai_identifier_str oai:digital.csic.es:10261/254808
network_acronym_str ES
network_name_str España
repository_id_str
spelling Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal FormArtusio, FioraCastellví, AlbertPisano, RobertoGavira Gallardo, J. A.ProteinCrystallizationPolymorphismAgarose gelNucleation© 2021 by the authors.Agarose gels provide the ideal environment for studying the nucleation step of complex biomacromolecules under diffusion-controlled conditions. In the present paper, we characterized the influence of agarose on the nucleation of three model proteins, i.e., lysozyme, insulin, and proteinase K, as a function of the agarose concentration using a batch method set-up inside flat capillaries. By using this set-up, we were able to directly count the number of crystals in a given volume and correlate it with the amount of agarose and with the average crystal size. We also studied the crystallization behavior of proteinase K with free-interface diffusion so that batch conditions were achieved through slow diffusion of the precipitant. Thanks to the control over the protein mass transport imposed by the network, a previously unknown crystal form, P212121, was obtained, and the three-dimensional structure was determined at a 1.6 Å resolution. Overall, the versatility of agarose gels makes them ideal candidates for the preparation of microcrystalline suspensions of biopharmaceuticals with precise and reproducible crystal attributes or for the exploration of the existence of different polymorphs.This research was funded by the project BIO2016-74875-P (MINECO), Spain, co-funded by the Fondo Europeo de Desarrollo Regional (FEDER funds), European Union.Peer reviewedMultidisciplinary Digital Publishing InstituteMinisterio de Economía y Competitividad (España)European CommissionConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202120212021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/254808reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-74875-Phttps://doi.org/10.3390/cryst11050466Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2548082026-05-22T06:33:51Z
dc.title.none.fl_str_mv Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form
title Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form
spellingShingle Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form
Artusio, Fiora
Protein
Crystallization
Polymorphism
Agarose gel
Nucleation
title_short Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form
title_full Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form
title_fullStr Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form
title_full_unstemmed Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form
title_sort Tuning Transport Phenomena in Agarose Gels for the Control of Protein Nucleation Density and Crystal Form
dc.creator.none.fl_str_mv Artusio, Fiora
Castellví, Albert
Pisano, Roberto
Gavira Gallardo, J. A.
author Artusio, Fiora
author_facet Artusio, Fiora
Castellví, Albert
Pisano, Roberto
Gavira Gallardo, J. A.
author_role author
author2 Castellví, Albert
Pisano, Roberto
Gavira Gallardo, J. A.
author2_role author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
European Commission
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Protein
Crystallization
Polymorphism
Agarose gel
Nucleation
topic Protein
Crystallization
Polymorphism
Agarose gel
Nucleation
description © 2021 by the authors.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/254808
url http://hdl.handle.net/10261/254808
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2016-74875-P
https://doi.org/10.3390/cryst11050466
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869425364020756480
score 15,81155

Similar Items