Institute of Atomic and Molecular Sciences, Academia Sinica, P.O. Box 23-166, Taipei 106, Taiwan.
Anal Chim Acta. 2010 Feb 5;659(1-2):201-7. doi: 10.1016/j.aca.2009.11.037. Epub 2009 Nov 22.
A solid-phase extraction method based on prestructured membrane filters and functionalized diamond nanocrystals is developed to facilitate analysis of biopolymers with MALDI mass spectrometry. The method integrates isolation, purification, and concentration in one step and is adaptable to high-throughput peptide fingerprint mapping of intact proteins and enzymatic digests. The feasibility of the method was demonstrated using diamond nanocrystals surface-derivatized with carboxylate groups and subsequently coated with poly-lysines. These nanocrystals, which are capable of extracting proteins from highly dilute solutions contaminated with salts and detergents, can be easily isolated by filtration through paraffin-coated PVDF membrane filters pre-patterned with channels for direct MALDI mass spectrometric analysis. This method is facile, easy to operate, and the analysis of each sample solution (with a volume up to 500 microL) can be completed in 10 min. With this method, it is possible to achieve high-throughput analysis of proteins by preparing an array of filtration spots on paraffin-coated PVDF membrane that matches the MALDI sample target of the commercial mass spectrometer.
一种基于预处理膜过滤器和功能化金刚石纳米晶体的固相萃取方法被开发出来，以促进 MALDI 质谱法分析生物聚合物。该方法集分离、纯化和浓缩于一步，适用于高通量的完整蛋白质和酶解物的肽指纹图谱分析。该方法使用表面衍生有羧酸盐的金刚石纳米晶体，并随后用聚赖氨酸涂覆，证明了其可行性。这些纳米晶体能够从含有盐和洗涤剂的高度稀释溶液中提取蛋白质，可以通过过滤预先用通道图案化的涂石蜡的聚偏二氟乙烯 (PVDF) 膜过滤器轻松分离，以便直接进行 MALDI 质谱分析。该方法简便易行，易于操作，每个样品溶液（体积可达 500 μL）的分析可在 10 分钟内完成。通过在涂石蜡的 PVDF 膜上制备与商用质谱仪的 MALDI 样品靶匹配的过滤斑点阵列，可以实现蛋白质的高通量分析。
