Peptide affinity chromatography based on combinatorial strategies for protein purification.
Methods Mol Biol
; 1129: 277-302, 2014.
Article
in En
| MEDLINE
| ID: mdl-24648083
ABSTRACT
We describe a method to develop affinity chromatography matrices with short peptide ligands for protein purification. The method entitles the following (a) synthesis of a combinatorial library on the hydromethylbenzoyl (HMBA)-ChemMatrix resin by the divide-couple-recombine (DCR) method using the Fmoc chemistry, (b) library screening with the protein of interest labeled with a fluorescent dye or biotin, (c) identification of peptides contained on positive beads by tandem matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS/MS), (d) solid-phase peptide ligand synthesis and immobilization in chromatographic supports, and (e) evaluation of protein adsorption on peptide affinity matrices from the equilibrium isotherms and breakthrough curves.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Peptides
/
Proteins
/
Chromatography, Affinity
Language:
En
Journal:
Methods Mol Biol
Journal subject:
BIOLOGIA MOLECULAR
Year:
2014
Document type:
Article