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

MARTINEZ CERON, MARIA CAMILA; GIUDICESSI, SILVANA L.; MARANI, MARIELA MIRTA; ALBERICIO, FERNANDO; CASCONE, OSVALDO; ERRA-BALSELLS, ROSA; CAMPERI, SILVIA ANDREA

ANALYTICAL BIOCHEMISTRY

ACADEMIC PRESS INC ELSEVIER SCIENCE

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

0003-2697

Different strategies for the identification of peptides selected after screening one-bead-one-peptide libraries were studied. This has been involved fine tuning of several steps of the whole process, which finish by MALDI MS. Guanidine, utilised for bead washing, was replaced by MeCN/AcOH/H2O, improving matrix crystallisation. Peptide-bead cleavage with NH4OH was cheaper, safer and as efficient NH3/THF. Peptides elution in microtubes instead of placing beads in the sample plate yielded more sample aliquots. “Successive-dry-layers deposit” sample preparation was better than “dried-droplet method”. Among the matrices analysed, a-cyano-4-hydroxycinnamic acid induced higher peptide ionisation. Clusters formation was minimised by additives addition to the matrix.