Department of Chemistry, National Sun Yat-Sen University, 70, Lien-Hai Road, Kaohsiung 80424, Taiwan.
J Mass Spectrom. 2010 Dec;45(12):1452-60. doi: 10.1002/jms.1861.
For the first time, we utilized multifunctional nanoparticles composite (NPs composite) for matrix-assisted laser desorption/ionization mass spectrometric (MALDI-MS) analysis of peptides and proteins. Multiwalled carbon nanotubes doped with Cd(2+) ions and modified with cadmium sulfide NPs were synthesized by a chemical reduction method at room temperature. The multifunctional NPs composite applied for the analysis of peptides and microwave-digested proteins in the atmospheric pressure matrix-assisted laser desorption/ionization ion-trap and MALDI time-of-flight (TOF) mass spectrometry (MS) was successfully demonstrated. The maximum detection sensitivity for peptides in MALDI-MS was achieved by the adsorption of negatively charged peptides onto the surfaces of NP composite through electrostatic interactions. The optimal conditions of peptide mixtures were obtained at 20 min of incubation time using 1 mg of NPs composite when the pH of the sample solution was kept higher than the pI values of peptides. The potentiality of the NP composite in the preconcentration of peptides was compared with that of the individual NP by calculating the preconcentration factors (PF) and found that the NPs composite showed a 4-6 times of PF than the other NPs. In addition, the NPs composite was also applied as heat-absorbing materials for efficient microwave tryptic digestion of cytochrome c and lysozyme from milk protein in MALDI-TOF-MS analysis. We believe that the use of NPs composite technique would be an efficient and powerful preconcentrating tool for MALDI-MS for the study of proteome research.
我们首次利用多功能纳米粒子复合材料(NPs 复合材料)进行肽和蛋白质的基质辅助激光解吸/电离质谱分析(MALDI-MS)。通过室温下的化学还原法合成了掺杂 Cd(2+)离子的多壁碳纳米管,并修饰有硫化镉 NPs。多功能 NPs 复合材料成功应用于大气压基质辅助激光解吸/电离离子阱和 MALDI 飞行时间(TOF)质谱(MS)中肽和微波消化蛋白质的分析。通过静电相互作用将带负电荷的肽吸附到 NPs 复合材料的表面,实现了 MALDI-MS 中对肽的最大检测灵敏度。在样品溶液的 pH 值高于肽的等电点时,在 20 分钟孵育时间内使用 1 mg NPs 复合材料,可以获得肽混合物的最佳条件。通过计算浓缩因子(PF)比较了 NPs 复合材料在肽浓缩方面的潜力与单个 NPs 的潜力,发现 NPs 复合材料的 PF 比其他 NPs 高 4-6 倍。此外,NPs 复合材料还被用作热吸收材料,用于 MALDI-TOF-MS 分析中从牛奶蛋白中高效微波消化细胞色素 c 和溶菌酶。我们相信,NPs 复合材料技术将成为 MALDI-MS 中研究蛋白质组学的有效、强大的浓缩工具。