Development of an affinity cryogel for one step purification of lysozyme from chicken egg white
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.jchromb.2016.12.032 http://hdl.handle.net/11449/174031 |
Resumo: | In this study, a supermacroporous polyacrylamide cryogel was produced by cryo-polymerization and activated with Tris(hydroxymethyl)aminomethane (Tris-cryogel) to be applied as an affinity ligand for a one step purification of lysozyme (LYZ), directly from chicken egg white (EW). The Tris-cryogel presented interconnected pores with size varying in the range of 20–80 μm and swelling capacity of 19.6 ± 0.9 g/g. The axial dispersion of the Tris-cryogel was analyzed at different flow velocities and mobile phase viscosities. It was verified that higher viscosity resulted in a higher degree of dispersion, causing the HETP values to increase from 0.04 cm to 0.8 cm. Adsorption isotherms were measured at 15 °C and 35 °C at pH 7.5. A Langmuir model was fitted to the equilibrium data, with a maximum adsorptive capacity of 285 mg/g at 15 °C and 363 mg/g at 35 °C. Thermodynamic analysis based on the Van't Hoff relationship showed that the process was spontaneous and enthalpically driven. Lysozyme was purified directly from egg white in a one step purification process at different pH values (7.5, 8.5 and 9.5). Independent of the pH, the specificity of Tris-cryogel for lysozyme adsorption was confirmed. At pH 7.5, yield and purification fold were higher (30% and 45). In addition, the effect of the dilution rate on egg white and flow velocity were also analyzed and it was shown that flow velocity did not affected purification and column efficiency, and that diluting the egg white increased yield to 70% with a purification fold of 23. Results show Tris-cryogel is a promising matrix for use in high throughput purification of lysozyme from egg white. |
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Repositório Institucional da UNESP |
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Development of an affinity cryogel for one step purification of lysozyme from chicken egg whiteAffinity chromatographyCryogelLysozymePurificationIn this study, a supermacroporous polyacrylamide cryogel was produced by cryo-polymerization and activated with Tris(hydroxymethyl)aminomethane (Tris-cryogel) to be applied as an affinity ligand for a one step purification of lysozyme (LYZ), directly from chicken egg white (EW). The Tris-cryogel presented interconnected pores with size varying in the range of 20–80 μm and swelling capacity of 19.6 ± 0.9 g/g. The axial dispersion of the Tris-cryogel was analyzed at different flow velocities and mobile phase viscosities. It was verified that higher viscosity resulted in a higher degree of dispersion, causing the HETP values to increase from 0.04 cm to 0.8 cm. Adsorption isotherms were measured at 15 °C and 35 °C at pH 7.5. A Langmuir model was fitted to the equilibrium data, with a maximum adsorptive capacity of 285 mg/g at 15 °C and 363 mg/g at 35 °C. Thermodynamic analysis based on the Van't Hoff relationship showed that the process was spontaneous and enthalpically driven. Lysozyme was purified directly from egg white in a one step purification process at different pH values (7.5, 8.5 and 9.5). Independent of the pH, the specificity of Tris-cryogel for lysozyme adsorption was confirmed. At pH 7.5, yield and purification fold were higher (30% and 45). In addition, the effect of the dilution rate on egg white and flow velocity were also analyzed and it was shown that flow velocity did not affected purification and column efficiency, and that diluting the egg white increased yield to 70% with a purification fold of 23. Results show Tris-cryogel is a promising matrix for use in high throughput purification of lysozyme from egg white.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)Department of Food Engineering and Technology UNESP- São Paulo State University, São José do Rio PretoDepartment of Food Science Federal University of Lavras, LavrasDepartment of Food Technology Federal University of Viçosa, ViçosaDepartment of Food Engineering and Technology UNESP- São Paulo State University, São José do Rio PretoUniversidade Estadual Paulista (Unesp)Federal University of LavrasFederal University of ViçosaMól, Paula Chequer Gouveia [UNESP]Veríssimo, Lizzy Ayra AlcântaraEller, Monique RenonMinim, Valéria Paula RodriguesMinim, Luis Antonio2018-12-11T17:08:49Z2018-12-11T17:08:49Z2017-02-15info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article17-23application/pdfhttp://dx.doi.org/10.1016/j.jchromb.2016.12.032Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, v. 1044-1045, p. 17-23.1873-376X1570-0232http://hdl.handle.net/11449/17403110.1016/j.jchromb.2016.12.0322-s2.0-850083509232-s2.0-85008350923.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences0,805info:eu-repo/semantics/openAccess2023-10-09T06:01:44Zoai:repositorio.unesp.br:11449/174031Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T14:23:48.242337Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Development of an affinity cryogel for one step purification of lysozyme from chicken egg white |
title |
Development of an affinity cryogel for one step purification of lysozyme from chicken egg white |
spellingShingle |
Development of an affinity cryogel for one step purification of lysozyme from chicken egg white Mól, Paula Chequer Gouveia [UNESP] Affinity chromatography Cryogel Lysozyme Purification |
title_short |
Development of an affinity cryogel for one step purification of lysozyme from chicken egg white |
title_full |
Development of an affinity cryogel for one step purification of lysozyme from chicken egg white |
title_fullStr |
Development of an affinity cryogel for one step purification of lysozyme from chicken egg white |
title_full_unstemmed |
Development of an affinity cryogel for one step purification of lysozyme from chicken egg white |
title_sort |
Development of an affinity cryogel for one step purification of lysozyme from chicken egg white |
author |
Mól, Paula Chequer Gouveia [UNESP] |
author_facet |
Mól, Paula Chequer Gouveia [UNESP] Veríssimo, Lizzy Ayra Alcântara Eller, Monique Renon Minim, Valéria Paula Rodrigues Minim, Luis Antonio |
author_role |
author |
author2 |
Veríssimo, Lizzy Ayra Alcântara Eller, Monique Renon Minim, Valéria Paula Rodrigues Minim, Luis Antonio |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Federal University of Lavras Federal University of Viçosa |
dc.contributor.author.fl_str_mv |
Mól, Paula Chequer Gouveia [UNESP] Veríssimo, Lizzy Ayra Alcântara Eller, Monique Renon Minim, Valéria Paula Rodrigues Minim, Luis Antonio |
dc.subject.por.fl_str_mv |
Affinity chromatography Cryogel Lysozyme Purification |
topic |
Affinity chromatography Cryogel Lysozyme Purification |
description |
In this study, a supermacroporous polyacrylamide cryogel was produced by cryo-polymerization and activated with Tris(hydroxymethyl)aminomethane (Tris-cryogel) to be applied as an affinity ligand for a one step purification of lysozyme (LYZ), directly from chicken egg white (EW). The Tris-cryogel presented interconnected pores with size varying in the range of 20–80 μm and swelling capacity of 19.6 ± 0.9 g/g. The axial dispersion of the Tris-cryogel was analyzed at different flow velocities and mobile phase viscosities. It was verified that higher viscosity resulted in a higher degree of dispersion, causing the HETP values to increase from 0.04 cm to 0.8 cm. Adsorption isotherms were measured at 15 °C and 35 °C at pH 7.5. A Langmuir model was fitted to the equilibrium data, with a maximum adsorptive capacity of 285 mg/g at 15 °C and 363 mg/g at 35 °C. Thermodynamic analysis based on the Van't Hoff relationship showed that the process was spontaneous and enthalpically driven. Lysozyme was purified directly from egg white in a one step purification process at different pH values (7.5, 8.5 and 9.5). Independent of the pH, the specificity of Tris-cryogel for lysozyme adsorption was confirmed. At pH 7.5, yield and purification fold were higher (30% and 45). In addition, the effect of the dilution rate on egg white and flow velocity were also analyzed and it was shown that flow velocity did not affected purification and column efficiency, and that diluting the egg white increased yield to 70% with a purification fold of 23. Results show Tris-cryogel is a promising matrix for use in high throughput purification of lysozyme from egg white. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017-02-15 2018-12-11T17:08:49Z 2018-12-11T17:08:49Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.jchromb.2016.12.032 Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, v. 1044-1045, p. 17-23. 1873-376X 1570-0232 http://hdl.handle.net/11449/174031 10.1016/j.jchromb.2016.12.032 2-s2.0-85008350923 2-s2.0-85008350923.pdf |
url |
http://dx.doi.org/10.1016/j.jchromb.2016.12.032 http://hdl.handle.net/11449/174031 |
identifier_str_mv |
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, v. 1044-1045, p. 17-23. 1873-376X 1570-0232 10.1016/j.jchromb.2016.12.032 2-s2.0-85008350923 2-s2.0-85008350923.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences 0,805 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
17-23 application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808128219179646976 |