title: Peptide affinity chromatography based on combinatorial strategies for protein purification.
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.
authors.affiliation: Institute of Nanobiotechnology UBA-CONICET, Cathedra of Industrial Microbiology and Biotechnology, School of Pharmacy and Biochemistry, University of Buenos Aires, Junín 956, C1113AAD, Buenos Aires, Argentina, scamperi@ffyb.uba.ar.
authors.forename: Silvia Andrea
authors.initials: SA
authors.lastname: Camperi
authors.forename: María Camila
authors.initials: MC
authors.lastname: Martínez-Ceron
authors.forename: Silvana Laura
authors.initials: SL
authors.lastname: Giudicessi
authors.forename: Mariela Mirta
authors.initials: MM
authors.lastname: Marani
authors.forename: Fernando
authors.initials: F
authors.lastname: Albericio
authors.forename: Osvaldo
authors.initials: O
authors.lastname: Cascone
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D020650:Combinatorial Chemistry Techniques; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000302:isolation & purification*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
chemical_list: D010455:Peptides; D011506:Proteins