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基质辅助激光解吸电离(MALDI)质谱:分子生物学和生物技术中的一种新型分析工具。

Matrix-assisted laser desorption ionization (MALDI) mass spectrometry: a novel analytical tool in molecular biology and biotechnology.

作者信息

Kaufmann R

机构信息

Institute of Laser Medicine and Biologisch-Medizinisches Forschungszentrum (BMFZ), Heinrich-Heine-University, Duesseldorf, Germany.

出版信息

J Biotechnol. 1995 Jul 31;41(2-3):155-75. doi: 10.1016/0168-1656(95)00009-f.

Abstract

Matrix-assisted laser desorption ionization (MALDI) is a rather recent 'soft ionization' technique which produces quasimolecular ions of large organic molecules of up to several 100 kDa molecular mass. In combination with time-of-flight (TOF) mass spectrometry it has rapidly evolved as a valuable tool for the detection and characterization of biopolymers such as peptides, proteins, oligosaccharides and -nucleotides especially in mixtures and crude samples. MALDI-MS is remarkably tolerant towards common contaminants such as salts and buffers, requires sample loads in the subpicomolar range and features a mass accuracy (and precision) of 0.1-0.01%. Structural information on analyte molecules can be obtained by exploiting metastable post source decay (PSD) processes which allow extraction of sequence or substituent information on individual peptides contained, e.g., in an enzymatic digest. Thus, MALDI provides for an easy and straight forward elucidation of posttranslational modifications. The forseeable merging of MALDI with blotting and in situ degradation techniques give MALDI-MS a key role in future laboratory practice of biotechnology and molecular biology. The actual state-of-the-art is described, and selected examples are given with special emphasis on the aspects of analytical routine work.

摘要

基质辅助激光解吸电离(MALDI)是一种相当新的“软电离”技术,它能产生分子量高达几百千道尔顿的大型有机分子的准分子离子。与飞行时间(TOF)质谱联用后,它迅速发展成为一种用于检测和表征生物聚合物(如肽、蛋白质、寡糖和核苷酸)的重要工具,尤其适用于混合物和粗样品。MALDI-MS对常见污染物(如盐和缓冲液)具有显著的耐受性,所需样品量在亚皮摩尔范围内,质量准确度(和精密度)为0.1 - 0.01%。通过利用亚稳的源后衰变(PSD)过程可以获得分析物分子的结构信息,该过程能够提取例如酶解消化产物中所含单个肽段的序列或取代基信息。因此,MALDI为翻译后修饰的轻松直接阐明提供了条件。MALDI与印迹和原位降解技术的可预见融合赋予了MALDI-MS在未来生物技术和分子生物学实验室实践中的关键作用。本文描述了当前的技术水平,并给出了选定的示例,特别强调了分析常规工作的各个方面。

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