Design of affinity chromatography peptide ligands through combinatorial peptide libraries screening

BARREDO G.R.; SAAVEDRA, SOLEDAD L.; MARTÍNEZ CERON MARÍA C.; GIUDICESSI SILVANA L.; MARANI MARIELA M.; ALBERICIO FERNANDO; CASCONE OSVALDO; CAMPERI SILVIA A.

Protein Downstream Processing: Design, Development and Application of High and Low-Resolution Methods

Springer

Lugar: Athens; Año: 2020; p. 217 - 244

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 hydroxymethylbenzoyl (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 ligands synthesis and immobilization on chromatographic supports, e) evaluation of protein adsorption on peptide affinity matrices through the equilibrium isotherms and breakthrough curves.