Cátedra de Microbiología Industrial y Biotecnología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, 1113 Buenos Aires, Argentina.
Anal Biochem. 2010 May 15;400(2):295-7. doi: 10.1016/j.ab.2010.01.029. Epub 2010 Feb 1.
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.
经过筛选单珠一单肽组合文库的优化，实现了基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）的珠分析优化，涉及整个过程的微调。用乙腈（MeCN）/乙酸（AcOH）/水（H(2)O）代替胍，改善了基质结晶。与 NH(3)/四氢呋喃（THF）相比，NH(4)OH 用于珠肽切割不仅便宜、安全，而且同样有效。与将珠放入样品板相比，在微量离心管中洗脱肽可得到更多的样品等分试样。连续的干层沉积样品制备优于干燥液滴法。在所分析的基质中，alpha-氰基-4-羟基肉桂酸产生了最佳的肽离子产率。通过向基质中添加添加剂，最小化了聚集体的形成。
