Simple method to assess stability of immobilized peptide ligands against proteases

Autores
Giudicessi, Silvana Laura; Salum, Maria Laura; Saavedra, Soledad Lorena; Martínez Ceron, María Camila; Cascone, Osvaldo; Erra Balsells, Rosa; Camperi, Silvia Andrea
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Although peptides are used as affinity chromatography ligands, they could be digested by proteases. Usually, peptide enzymatic stability is evaluated in solution, which differs from the resin-bounded peptide behavior. Furthermore, the study of the degradation products requires purification steps before analysis. Here we describe an easy method to evaluate immobilized peptide stability. Sample peptides were synthesized on hydroxymethylbenzamide-ChemMatrix resin. Peptidyl-resin beads were then incubated with solutions containing proteases. Peptides were detached from the solid support with ammonia vapor and analyzed by MALDI and ESI mass spectrometry, allowing the detection of the whole peptide as well as their C-terminal degradation products. The method allowed a fast evaluation of peptide ligands stability in solid phase towards proteases that may be present in the crude sample before their use in affinity chromatography.
Fil: Giudicessi, Silvana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina
Fil: Salum, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Orgánica; Argentina
Fil: Saavedra, Soledad Lorena. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina
Fil: Martínez Ceron, María Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina
Fil: Cascone, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina
Fil: Erra Balsells, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Orgánica; Argentina
Fil: Camperi, Silvia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina
Materia
Mass Spectrometry
Peptides
Amino Acids
Amino Acids Derivatives
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/47466
Acceder
id CONICETDig_209a3455199aca54c805554dc0917ae3
oai_identifier_str oai:ri.conicet.gov.ar:11336/47466
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Simple method to assess stability of immobilized peptide ligands against proteasesGiudicessi, Silvana LauraSalum, Maria LauraSaavedra, Soledad LorenaMartínez Ceron, María CamilaCascone, OsvaldoErra Balsells, RosaCamperi, Silvia AndreaMass SpectrometryPeptidesAmino AcidsAmino Acids Derivativeshttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Although peptides are used as affinity chromatography ligands, they could be digested by proteases. Usually, peptide enzymatic stability is evaluated in solution, which differs from the resin-bounded peptide behavior. Furthermore, the study of the degradation products requires purification steps before analysis. Here we describe an easy method to evaluate immobilized peptide stability. Sample peptides were synthesized on hydroxymethylbenzamide-ChemMatrix resin. Peptidyl-resin beads were then incubated with solutions containing proteases. Peptides were detached from the solid support with ammonia vapor and analyzed by MALDI and ESI mass spectrometry, allowing the detection of the whole peptide as well as their C-terminal degradation products. The method allowed a fast evaluation of peptide ligands stability in solid phase towards proteases that may be present in the crude sample before their use in affinity chromatography.Fil: Giudicessi, Silvana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: Salum, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Orgánica; ArgentinaFil: Saavedra, Soledad Lorena. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Martínez Ceron, María Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: Cascone, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaFil: Erra Balsells, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Orgánica; ArgentinaFil: Camperi, Silvia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; ArgentinaJohn Wiley & Sons Ltd2017-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/47466Giudicessi, Silvana Laura; Salum, Maria Laura; Saavedra, Soledad Lorena; Martínez Ceron, María Camila; Cascone, Osvaldo; et al.; Simple method to assess stability of immobilized peptide ligands against proteases; John Wiley & Sons Ltd; Journal Of Peptide Science; 23; 9; 9-2017; 685-6921075-2617CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1002/psc.3012info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1002/psc.3012info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:48:04Zoai:ri.conicet.gov.ar:11336/47466instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:48:04.866CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Simple method to assess stability of immobilized peptide ligands against proteases
title Simple method to assess stability of immobilized peptide ligands against proteases
spellingShingle Simple method to assess stability of immobilized peptide ligands against proteases
Giudicessi, Silvana Laura
Mass Spectrometry
Peptides
Amino Acids
Amino Acids Derivatives
title_short Simple method to assess stability of immobilized peptide ligands against proteases
title_full Simple method to assess stability of immobilized peptide ligands against proteases
title_fullStr Simple method to assess stability of immobilized peptide ligands against proteases
title_full_unstemmed Simple method to assess stability of immobilized peptide ligands against proteases
title_sort Simple method to assess stability of immobilized peptide ligands against proteases
dc.creator.none.fl_str_mv Giudicessi, Silvana Laura
Salum, Maria Laura
Saavedra, Soledad Lorena
Martínez Ceron, María Camila
Cascone, Osvaldo
Erra Balsells, Rosa
Camperi, Silvia Andrea
author Giudicessi, Silvana Laura
author_facet Giudicessi, Silvana Laura
Salum, Maria Laura
Saavedra, Soledad Lorena
Martínez Ceron, María Camila
Cascone, Osvaldo
Erra Balsells, Rosa
Camperi, Silvia Andrea
author_role author
author2 Salum, Maria Laura
Saavedra, Soledad Lorena
Martínez Ceron, María Camila
Cascone, Osvaldo
Erra Balsells, Rosa
Camperi, Silvia Andrea
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Mass Spectrometry
Peptides
Amino Acids
Amino Acids Derivatives
topic Mass Spectrometry
Peptides
Amino Acids
Amino Acids Derivatives
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Although peptides are used as affinity chromatography ligands, they could be digested by proteases. Usually, peptide enzymatic stability is evaluated in solution, which differs from the resin-bounded peptide behavior. Furthermore, the study of the degradation products requires purification steps before analysis. Here we describe an easy method to evaluate immobilized peptide stability. Sample peptides were synthesized on hydroxymethylbenzamide-ChemMatrix resin. Peptidyl-resin beads were then incubated with solutions containing proteases. Peptides were detached from the solid support with ammonia vapor and analyzed by MALDI and ESI mass spectrometry, allowing the detection of the whole peptide as well as their C-terminal degradation products. The method allowed a fast evaluation of peptide ligands stability in solid phase towards proteases that may be present in the crude sample before their use in affinity chromatography.
Fil: Giudicessi, Silvana Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina
Fil: Salum, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Orgánica; Argentina
Fil: Saavedra, Soledad Lorena. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina
Fil: Martínez Ceron, María Camila. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina
Fil: Cascone, Osvaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina
Fil: Erra Balsells, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Orgánica; Argentina
Fil: Camperi, Silvia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Nanobiotecnología. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Nanobiotecnología; Argentina
description Although peptides are used as affinity chromatography ligands, they could be digested by proteases. Usually, peptide enzymatic stability is evaluated in solution, which differs from the resin-bounded peptide behavior. Furthermore, the study of the degradation products requires purification steps before analysis. Here we describe an easy method to evaluate immobilized peptide stability. Sample peptides were synthesized on hydroxymethylbenzamide-ChemMatrix resin. Peptidyl-resin beads were then incubated with solutions containing proteases. Peptides were detached from the solid support with ammonia vapor and analyzed by MALDI and ESI mass spectrometry, allowing the detection of the whole peptide as well as their C-terminal degradation products. The method allowed a fast evaluation of peptide ligands stability in solid phase towards proteases that may be present in the crude sample before their use in affinity chromatography.
publishDate 2017
dc.date.none.fl_str_mv 2017-09
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/47466
Giudicessi, Silvana Laura; Salum, Maria Laura; Saavedra, Soledad Lorena; Martínez Ceron, María Camila; Cascone, Osvaldo; et al.; Simple method to assess stability of immobilized peptide ligands against proteases; John Wiley & Sons Ltd; Journal Of Peptide Science; 23; 9; 9-2017; 685-692
1075-2617
CONICET Digital
CONICET
url http://hdl.handle.net/11336/47466
identifier_str_mv Giudicessi, Silvana Laura; Salum, Maria Laura; Saavedra, Soledad Lorena; Martínez Ceron, María Camila; Cascone, Osvaldo; et al.; Simple method to assess stability of immobilized peptide ligands against proteases; John Wiley & Sons Ltd; Journal Of Peptide Science; 23; 9; 9-2017; 685-692
1075-2617
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1002/psc.3012
info:eu-repo/semantics/altIdentifier/url/https://onlinelibrary.wiley.com/doi/abs/10.1002/psc.3012
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv John Wiley & Sons Ltd
publisher.none.fl_str_mv John Wiley & Sons Ltd
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1844613495244455936
score 13.070432

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