INVESTIGADORES
CAMPERI Silvia Andrea
capítulos de libros
Título:
Design of affinity chromatography peptide ligands through combinatorial peptide libraries screening
Autor/es:
G. R. BARREDO; S. L. SAAVEDRA; M. C. MARTÍNEZ CERON; S. L. GIUDICESSI; M. M. MARANI; F. ALBERICIO; O. CASCONE; S. A. CAMPERI
Libro:
Springer book Methods in Molecular Biology: 2nd Edition of Protein Downstream Processing
Editorial:
Springer
Referencias:
Año: 2021; p. 217 - 243
Resumen:
In this chapter a protocol to design affinity chromatography matrices with short peptide ligands immobilized for protein purification is described. The first step consists on the synthesis of a combinatorial peptide library on the hydromethylbenzoyl (HMBA)-ChemMatrix resin by the divide-couple-recombine (DCR) method using the Fmoc chemistry. Next, the library is screened with the protein of interest labeled with a fluorescent dye or biotin. Subsequently, peptides contained on positive beads are identificated by tandem matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS/MS) and those sequences showing greater consensus are synthesized in larger quantities and immobilized on chromatographic supports. Finally, target protein adsorption on peptide affinity matrices are evaluated through equilibrium adsorption isotherms and breakthrough curves.
