Simple method to assess stability of immobilized peptide ligands against proteases
Although peptides are used as affinity chromatography ligands, they could be digested by proteases. Usually, peptide stability is evaluated in solution, which differs from the resin-bounded peptide behavior. Furthermore, the study of the degradation products requires purification steps before analys...
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2017
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10752617_v23_n9_p685_Giudicessi http://hdl.handle.net/20.500.12110/paper_10752617_v23_n9_p685_Giudicessi |
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paper:paper_10752617_v23_n9_p685_Giudicessi2023-06-08T16:05:12Z Simple method to assess stability of immobilized peptide ligands against proteases Giudicessi, Silvana Laura Erra Balsells, Rosa affinity chromatography ESI MS MALDI MS purification ammonia benzamide derivative immobilized protein ligand proteinase resin peptide peptide hydrolase affinity chromatography analytic method Article carboxy terminal sequence matrix-assisted laser desorption-ionization mass spectrometry peptide synthesis priority journal protein degradation solid chemistry metabolism Chromatography, Affinity Peptide Hydrolases Peptides Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Although peptides are used as affinity chromatography ligands, they could be digested by proteases. Usually, peptide 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 assess 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 matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry, allowing the detection of the whole peptides as well as their C-terminal degradation products. The method allowed a fast evaluation of peptide ligand stability in solid phase towards proteases that may be present in the crude sample before their use as ligands in affinity chromatography. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Fil:Giudicessi, S.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Erra-Balsells, R. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2017 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10752617_v23_n9_p685_Giudicessi http://hdl.handle.net/20.500.12110/paper_10752617_v23_n9_p685_Giudicessi |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
affinity chromatography ESI MS MALDI MS purification ammonia benzamide derivative immobilized protein ligand proteinase resin peptide peptide hydrolase affinity chromatography analytic method Article carboxy terminal sequence matrix-assisted laser desorption-ionization mass spectrometry peptide synthesis priority journal protein degradation solid chemistry metabolism Chromatography, Affinity Peptide Hydrolases Peptides Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization |
| spellingShingle |
affinity chromatography ESI MS MALDI MS purification ammonia benzamide derivative immobilized protein ligand proteinase resin peptide peptide hydrolase affinity chromatography analytic method Article carboxy terminal sequence matrix-assisted laser desorption-ionization mass spectrometry peptide synthesis priority journal protein degradation solid chemistry metabolism Chromatography, Affinity Peptide Hydrolases Peptides Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Giudicessi, Silvana Laura Erra Balsells, Rosa Simple method to assess stability of immobilized peptide ligands against proteases |
| topic_facet |
affinity chromatography ESI MS MALDI MS purification ammonia benzamide derivative immobilized protein ligand proteinase resin peptide peptide hydrolase affinity chromatography analytic method Article carboxy terminal sequence matrix-assisted laser desorption-ionization mass spectrometry peptide synthesis priority journal protein degradation solid chemistry metabolism Chromatography, Affinity Peptide Hydrolases Peptides Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization |
| description |
Although peptides are used as affinity chromatography ligands, they could be digested by proteases. Usually, peptide 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 assess 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 matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry, allowing the detection of the whole peptides as well as their C-terminal degradation products. The method allowed a fast evaluation of peptide ligand stability in solid phase towards proteases that may be present in the crude sample before their use as ligands in affinity chromatography. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. |
| author |
Giudicessi, Silvana Laura Erra Balsells, Rosa |
| author_facet |
Giudicessi, Silvana Laura Erra Balsells, Rosa |
| author_sort |
Giudicessi, Silvana Laura |
| title |
Simple method to assess stability of immobilized peptide ligands against proteases |
| 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 |
| publishDate |
2017 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10752617_v23_n9_p685_Giudicessi http://hdl.handle.net/20.500.12110/paper_10752617_v23_n9_p685_Giudicessi |
| work_keys_str_mv |
AT giudicessisilvanalaura simplemethodtoassessstabilityofimmobilizedpeptideligandsagainstproteases AT errabalsellsrosa simplemethodtoassessstabilityofimmobilizedpeptideligandsagainstproteases |
| _version_ |
1768545660740567040 |