Sample preparation for sequencing hits from one-bead-one-peptide combinatorial libraries by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI application)

MARÍA C. MARTÍNEZ CERÓN; SILVANA L. GIUDICESSI; MARIELA M. MARANI; FERNANDO ALBERICIO; OSVALDO CASCONE; ROSA ERRA-BALSELLS; SILVIA A. CAMPERI

ANALYTICAL BIOCHEMISTRY

ACADEMIC PRESS INC ELSEVIER SCIENCE

Lugar: Amsterdam; Año: 2010 vol. 400 p. 295 - 297

0003-2697

Optimization of bead analysis by matrix-assisted laser desoprtion/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) after the screening of one-bead-one-peptide combinatorial libraries was achieved, involving the fine-tuning of the whole process. Guanidine was replaced by acetonitrile (MeCN)/acetic acid (AcOH)/water (H2O), improving matrix crystallization. Peptide-bead cleavage  with NH4OH was cheaper and safer than, yet as efficient as, NH3/tetrahydrofuran (THF). Peptide elution in microtubes instead of placing the beads in the sample plate yielded more sample aliquots. Successive dry layers deposit sample preparation was better than the dried droplet method. Among yhe matrices analyzed, A-ciano-4-hydroxycinnamic acid resulted in the best peptide ion yield. Cluster formation was minimezed by the addition of additives to the matrix