Peptide affinity chromatography based on combinatorial strategies for protein purification.

S.A. CAMPERI; M. C. MARTÍNEZ-CERON; S. L. GIUDICESSI; M.M. MARANI; F. ALBERICIO; O. CASCONE

Methods in Molecular Biology. Protein Downstream Processing.

Humana Press-Springer

Lugar: Nueva York; Año: 2014; p. 277 - 302

Here we describe a method to developed affinity chromatography matrices with short peptide ligands for protein purification. The method entitles the following: a) synthesis of a combinatorial library on the HMBA- ChemMatrix resin by the divide-couple-recombine (DCR) method using Fmoc chemistry, b) library screening with the protein of interested labeled with a fluorescent dye or biotin. c) identification of peptides contained on positive beads by tandem matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS/MS), d) solid phase peptide ligands synthesis and immobilization in chromatographic supports, e) evaluation of protein adsorption on peptide affinity matrices by the constructions of equilibrium isotherms and breakthroughs.