Sample preparation for sequencing hits from one-bead-one-peptide combinatorial libraries by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.
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 (H(2)O), improving matrix crystallization. Peptide-bead cleavage with NH(4)OH was cheaper and safer than, yet as efficient as, NH(3)/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, alpha-cyano-4-hydroxycinnamic acid resulted in the best peptide ion yield. Cluster formation was minimized by the addition of additives to the matrix.
Martínez-Ceron
María C
MC
Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, 1113 Buenos Aires, Argentina.
Giudicessi
Silvana L
SL
Marani
Mariela M
MM
Albericio
Fernando
F
Cascone
Osvaldo
O
Erra-Balsells
Rosa
R
Camperi
Silvia A
SA
D019342:Acetic Acid / Q000737:chemistry; D000097:Acetonitriles / Q000737:chemistry; D064753:Ammonium Hydroxide; D006878:Hydroxides / Q000737:chemistry; D019151:Peptide Library; D010455:Peptides / Q000032:analysis* / Q000737:chemistry; D020539:Sequence Analysis, Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014867:Water / Q000737:chemistry
D000097:Acetonitriles; D006878:Hydroxides; D019151:Peptide Library; D010455:Peptides; D014867:Water; D064753:Ammonium Hydroxide; D019342:Acetic Acid; C032159:acetonitrile