Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents

High-performance frontal affinity chromatography was employed to study the adsorption–desorption kinetics characterizing the retention of Beta-lactoglobulin (Beta-LG) onto polyclonal anti-Beta-lactoglobulin (anti-Beta-LG) chromatographic supports. The adsorption and desorption processes were studie...

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Detalles Bibliográficos
Autores: Puerta, Angel de la, Vidal-Madjar, Claire, Jaulmes, Alain, Díez-Masa, José Carlos, Frutos, Mercedes de
Tipo de recurso: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2006
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/40403
Acceso en línea:http://hdl.handle.net/10261/40403
Access Level:acceso abierto
Palabra clave:Computer simulation
Adsorption–desorption kinetics
Affinity chromatography
Lactoglobulin
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spelling Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbentsPuerta, Angel de laVidal-Madjar, ClaireJaulmes, AlainDíez-Masa, José CarlosFrutos, Mercedes deComputer simulationAdsorption–desorption kineticsAffinity chromatographyLactoglobulinHigh-performance frontal affinity chromatography was employed to study the adsorption–desorption kinetics characterizing the retention of Beta-lactoglobulin (Beta-LG) onto polyclonal anti-Beta-lactoglobulin (anti-Beta-LG) chromatographic supports. The adsorption and desorption processes were studied by analyzing two different elution fronts separated by a relatively long rinsing step. The method consists in performing two successive frontal injections of the protein. In between, the column was rinsed with a given volume of mobile phase (buffer alone). During this rinsing stage, a partial desorption may occur and a novel amount of protein could be adsorbed in the second frontal injection step. The whole process (first adsorption, possible desorption, and second adsorption) was simulated by a numerical procedure, in which the column was divided into a large number of slices. A model based on bi-Langmuir type kinetics was used to describe the adsorption of the protein on the support. The model assumes a non-uniform adsorbent with two types of binding sites. At equilibrium the adsorption isotherm is of the bi-Langmuir type. A global adsorption effect was considered which includes the effective binding process and the mass transfer resistances due to the transport to the binding site. Therefore, the column capacity and the kinetic parameters of the model (apparent adsorption and desorption rate constants) were determined from the best fit of the first and second adsorption fronts to the experimental ones. The other parameters of the model are the saturation capacities for the adsorption on each type of sites. The equilibrium affinity constants were estimated in a single experiment from the ratio of the apparent adsorption and desorption rate constants. The high values found (around 108 M−1) reveal a strong interaction of -LG with the immunoadsorbent. Kinetic measurements were carried out at different flow rates. Both the apparent adsorption and desorption kinetics were faster at larger flow rates, indicating an important contribution of the mass transfer resistance in the stagnant fluid at the particle boundary. However, as expected, close values were found for the resulting equilibrium constants calculated from the ratio of the apparent adsorption and desorption rate constant determined at various flow rates.A.P. acknowledges Spanish Ministry of Science and Tech- nology for a predoctoral grant. This work has been supported by Spanish CICYT (Project TIC2003-01906) and Foundation Ramon Areces. Collaboration between laboratories has been possible thanks to French-Spanish Cooperation between CNRS and CSIC.Peer reviewedElsevierConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]201120112006info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Postprintinfo:eu-repo/semantics/acceptedVersionhttp://hdl.handle.net/10261/40403reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Ingléshttp://dx.doi.org/10.1016/j.chroma.2005.12.010Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/404032026-05-22T06:33:51Z
dc.title.none.fl_str_mv Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents
title Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents
spellingShingle Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents
Puerta, Angel de la
Computer simulation
Adsorption–desorption kinetics
Affinity chromatography
Lactoglobulin
title_short Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents
title_full Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents
title_fullStr Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents
title_full_unstemmed Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents
title_sort Frontal analysis for characterizing the adsorption-desorption behavior of beta-lactoglobulin on immunoadsorbents
dc.creator.none.fl_str_mv Puerta, Angel de la
Vidal-Madjar, Claire
Jaulmes, Alain
Díez-Masa, José Carlos
Frutos, Mercedes de
author Puerta, Angel de la
author_facet Puerta, Angel de la
Vidal-Madjar, Claire
Jaulmes, Alain
Díez-Masa, José Carlos
Frutos, Mercedes de
author_role author
author2 Vidal-Madjar, Claire
Jaulmes, Alain
Díez-Masa, José Carlos
Frutos, Mercedes de
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Computer simulation
Adsorption–desorption kinetics
Affinity chromatography
Lactoglobulin
topic Computer simulation
Adsorption–desorption kinetics
Affinity chromatography
Lactoglobulin
description High-performance frontal affinity chromatography was employed to study the adsorption–desorption kinetics characterizing the retention of Beta-lactoglobulin (Beta-LG) onto polyclonal anti-Beta-lactoglobulin (anti-Beta-LG) chromatographic supports. The adsorption and desorption processes were studied by analyzing two different elution fronts separated by a relatively long rinsing step. The method consists in performing two successive frontal injections of the protein. In between, the column was rinsed with a given volume of mobile phase (buffer alone). During this rinsing stage, a partial desorption may occur and a novel amount of protein could be adsorbed in the second frontal injection step. The whole process (first adsorption, possible desorption, and second adsorption) was simulated by a numerical procedure, in which the column was divided into a large number of slices. A model based on bi-Langmuir type kinetics was used to describe the adsorption of the protein on the support. The model assumes a non-uniform adsorbent with two types of binding sites. At equilibrium the adsorption isotherm is of the bi-Langmuir type. A global adsorption effect was considered which includes the effective binding process and the mass transfer resistances due to the transport to the binding site. Therefore, the column capacity and the kinetic parameters of the model (apparent adsorption and desorption rate constants) were determined from the best fit of the first and second adsorption fronts to the experimental ones. The other parameters of the model are the saturation capacities for the adsorption on each type of sites. The equilibrium affinity constants were estimated in a single experiment from the ratio of the apparent adsorption and desorption rate constants. The high values found (around 108 M−1) reveal a strong interaction of -LG with the immunoadsorbent. Kinetic measurements were carried out at different flow rates. Both the apparent adsorption and desorption kinetics were faster at larger flow rates, indicating an important contribution of the mass transfer resistance in the stagnant fluid at the particle boundary. However, as expected, close values were found for the resulting equilibrium constants calculated from the ratio of the apparent adsorption and desorption rate constant determined at various flow rates.
publishDate 2006
dc.date.none.fl_str_mv 2006
2011
2011
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Postprint
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/40403
url http://hdl.handle.net/10261/40403
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv http://dx.doi.org/10.1016/j.chroma.2005.12.010
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
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
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