Nguyen-Khuong Terry, Fitzgerald Anna, Zhao Zhejun, Willcox Mark, Walsh Bradley J
Minomic Pty Ltd, Frenchs Forest, NSW, Australia.
Proteomics. 2008 Sep;8(17):3424-32. doi: 10.1002/pmic.200701161.
Many low-molecular weight proteins and peptides in human tears are potentially bioactive proteins but the range and number of these is yet to be fully characterized. A number of different sample preparation techniques were used to maximize the visualization of peptides from reflex tears. Samples were pretreated using precipitation and filtration techniques prior to analyses using MALDI-TOF MS. Peptides were searched for between 700 to 4000 m/z. Sample dilution in several different buffer systems followed by filtration with a 30-kDa cutoff filter and C18 reverse phase microcolumn purification produced significantly (p = 0.049) more peaks in tears than other methods used to prepare tears prior to MALDI-TOF MS. This study has established a technique for optimizing the visualization of naturally occurring peptides in tears.
人泪液中的许多低分子量蛋白质和肽可能是生物活性蛋白，但它们的种类和数量尚未完全明确。使用了多种不同的样品制备技术，以最大限度地观察反射性泪液中的肽。在使用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）分析之前，样品先采用沉淀和过滤技术进行预处理。在700至4000 m/z之间搜索肽。与用于在MALDI-TOF MS分析之前制备泪液的其他方法相比，在几种不同缓冲系统中进行样品稀释，然后用截留分子量为30 kDa的滤膜过滤并通过C18反相微柱纯化，能显著（p = 0.049）产生更多的泪液峰。本研究建立了一种优化泪液中天然存在肽可视化的技术。
