官能化奈米粒子應用於辨認生化分子之親和探針及結合溶劑微萃取進行胜肽與蛋白質的質譜分析

Applications of Functional Nanoparticles as the Affinity Probes for Analysis of Peptides and Proteins by Coupling Solvent Microextraction with the MALDI Mass Spectrometry

柯耀棠(Yao-Tang Ke) ； 吳慧芬(Hui-Fen Wu)

《化學》 66卷3期 (2008/09) Pp. 231-240

https://doi.org/10.6623/chem.2008023

摘要

基質輔助雷射脫附游離質譜法(MALDI)為目前常被用於分析生化樣品，然而在此質譜法中使用傳統有機基質，會造成在低質量範圍的背景干擾，因而限制了它於小分子的分析適用性。此外，在產生共結晶和非共結晶時會有不均勻化的情形發生，產生集中點(sweet spot)的問題。目前已有文獻報導使用奈米材料可改善此問題，該方法稱為SALDI (Surfaced Assisted Laser Desorption Ionization)。本實驗室近年來，首度開發出奈米粒子在液相微萃取之應用；亦開發一滴溶劑微萃取法作為基質輔助雷射脫附質譜法之濃縮探針以提高生化分子偵測的靈敏度。本篇報導主要介紹我們實驗室結合液相微萃取和奈米粒子的分析技術，此外也開發奈米粒子的表面特性作為預濃縮探針，進行有效地快速分析胺硫類(aminothiol)、蛋白質(protein)、胜肽混合(peptide mixtures)的分析。本篇報導分別以金奈米粒子，銀奈米粒子結合溶劑萃取法進行蛋白質(protein)和胜肽(peptide)的分析。此外也利用二氧化矽奈米粒子(SiO_2 nanoparticles)的表面電荷關係，有效地把樣品濃縮出來，而來進行分析。

關鍵字

基質輔助雷射脫附游離法 ； 金奈米粒子 ； 二氧化矽奈米粒子 ； 銀奈米粒子 ； 萃取

並列摘要

Matrix-Aassisted Laser Desorption/Ionization mass spectrometry (MALDI-MS) is a powerful tool for the analysis of biomolecules such as peptides and proteins and soft ionization technique using the organic matrix. Because of the high concentration of the organic matrix produces high background signals in the low mass range, nanoparticles have been intensively applied in the Surface-Assisted Laser Desorption/Ionization-Mass Spectrometry (SALDI-MS) to reduce the background interferences in the MALDI-MS. Recently, our group has developed a novel approach of using modified gold or silver nanoparticles (AgNPs) prepared in toluene as electrostatic or hydrophobic affinity probes for the separation and preconcentration of peptides and proteins in biological samples prior to atmospheric pressure- matrix assisted laser desorption/ionization (AP-MALDI) ion trap mass spectrometry and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry. To our best knowledge, this is the first time that modified Au or AgNPs were prepared in organic solvent and also combined the liquid-phase microextraction (LPME) techniques to analyze peptides and proteins in the MALDI-MS. In the present report, we introduced the proposed method for efficient extraction and rapid enrichment of small thio-amino acids, peptides and large proteins like, myoglobin, ubiquitin and bovine serum albumin in biological sample such as urine and plasma by matrix assisted laser desorption/ionization mass spectrometry (MALDI-MS). The advantages of this technique are simple, fast, sensitive and efficient. It is an ideal and preconcentrating method prior to MALDI-MS detection for analysis of peptides and proteins from biological samples.

並列關鍵字

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) ； Gold nanoparticles ； Silica nanoparticles ； Silver nanoparticles ； Extraction