Department of Chemistry, Materials and Chemical Engineering Giulio Natta, Via Mancinelli 7, Politecnico di Milano, Milano, Italy.
Expert Rev Proteomics. 2010 Jun;7(3):373-85. doi: 10.1586/epr.10.25.
The latest advances in combinatorial peptide ligand libraries, with their unique performance in discovering low-abundance species in proteomes, are reviewed here. Explanations of mechanism, potential applications, capture of proteomes at different pH values to enhance the total catch and quantitative elutions, such as boiling in the presence of 5% sodium dodecyl sulfate and 3% dithiothreitol are included. The reproducibility of protein capture among different experiments with the same batch of beads or with different batches is also reported to be very high, with coefficient of variations in the order of 10-20%. Miniaturized operations, consisting of capture with as little as 20 or even 5 microl of peptide beads are reported, thus demonstrating that the described technology could be exploited for routine biomarker discovery in a biomedical environment. Finally, it is shown that the signal of captured proteins is linear over approximately three orders of magnitude, ranging from nM to microM, thus ensuring that differential quantitative proteomics for biomarker discovery can be fully implemented, providing species do not saturate their ligands.
本文综述了组合肽配体文库的最新进展，它们在发现蛋白质组中低丰度物种方面具有独特的性能。文中解释了其作用机制、潜在应用，以及在不同 pH 值下捕获蛋白质组以增强总捕获量和定量洗脱的方法，如在存在 5%十二烷基硫酸钠和 3%二硫苏糖醇的情况下煮沸。还报告了使用同一批或不同批珠子进行的不同实验中蛋白质捕获的重复性非常高，变异系数在 10-20%左右。报道称，微型操作包括用 20 甚至 5 微升肽珠进行捕获，这表明该技术可用于生物医学环境中的常规生物标志物发现。最后，结果表明，捕获蛋白的信号在线性范围内跨越约三个数量级，从 nM 到 microM，从而确保可以完全实施用于生物标志物发现的差异定量蛋白质组学，只要物种不会使它们的配体饱和。
