Sample preparation for sequencing hits from one-bead-one-peptide combinatorial libraries by MALDI MS

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

ANALYTICAL BIOCHEMISTRY

Elsevier

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

0003-2697

Optimization of bead analysis by matrix-assisted laser desorption/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 the matrices analyzed, a-cyano-4-hydroxycinnamic acid resulted in the best peptide ion yield. Cluster formation was minimized by the addition of additives to the matrix.