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Simple method to assess stability of immobilized peptide ligands against proteases
Author(s) -
Giudicessi Silvana L.,
Salum María L.,
Saavedra Soledad L.,
MartínezCeron María C.,
Cascone Osvaldo,
ErraBalsells Rosa,
Camperi Silvia A.
Publication year - 2017
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.3012
Subject(s) - proteases , peptide , chemistry , chromatography , electrospray ionization , mass spectrometry , matrix assisted laser desorption/ionization , desorption , affinity chromatography , combinatorial chemistry , enzyme , biochemistry , organic chemistry , adsorption
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.