1. Simple method to assess stability of immobilized peptide ligands against proteases
- Author
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María L. Salum, María C. Martínez-Ceron, Osvaldo Cascone, Soledad L. Saavedra, Rosa Erra-Balsells, Silvia A. Camperi, and Silvana L. Giudicessi
- Subjects
Pharmacology ,chemistry.chemical_classification ,Proteases ,Chromatography ,Protein mass spectrometry ,010405 organic chemistry ,Chemistry ,Electrospray ionization ,010401 analytical chemistry ,Organic Chemistry ,Peptide ,General Medicine ,Mass spectrometry ,01 natural sciences ,Biochemistry ,Combinatorial chemistry ,0104 chemical sciences ,Amino acid ,Affinity chromatography ,Structural Biology ,Desorption ,Drug Discovery ,Molecular Medicine ,Molecular Biology - Abstract
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.
- Published
- 2017
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