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首页> 外文会议>ASMS Conference on Mass Spectrometry and Allied Topics >Reactive Electrospray-Assisted Laser Desorption Ionization (Reactive ELDI) for Characterization of Peptides and Proteins
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Reactive Electrospray-Assisted Laser Desorption Ionization (Reactive ELDI) for Characterization of Peptides and Proteins

机译：反应电喷雾辅助激光解吸电离（反应性ELDI），用于表征肽和蛋白质

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ELDI is a soft ionization method which combines features of both ESI and MALDI. ELDI works under ambient conditions for protein samples deposited on a sample plate. The ELDI ionization process consists of two separate steps: generation of gaseous sample molecules by laser desorption, and subsequent post-ionization of the sample particles by ESI. ELDI generates multiply charged ions, which is amenable for top-down protein identification with various MS/MS techniques (Peng et al. "Electrospray-Assisted Laser Desorption Ionization (ELDI) and Tandem Mass Spectrometry of Peptides and Proteins." Rapid Commun Mass Spectrom 2007; 21: 2541-2546). ELDI is shown to be capable of characterizing large proteins up to 29 kDa. Multiply charged proteins generated by the ELDI mechanism can be shifted to higher charge state either by decreasing the pH, or by adding m-nitrobenzyl alcohol (m-NBA) in the ESI solution. Reactive ELDI allows chemical reactions to occur during the ELDI process. Reactive ELDI experiments are shown to be effective for gas phase formation of ribonuclease A (RNase A)-CTP complex, and for on-line disulfide reduction with dithiothreitol (DTT) for oxidized glutathione and insulin. These data provide a possibility for direct protein-ligand binding study, as well as on-line top-down protein identification for proteomic study, without the necessity of laborious sample preparation.
机译：ELDI是结合了ESI和MALDI的特征的软电离方法。 ELDI适用于沉积在样品板的蛋白质样品在环境条件下。所述ELDI电离过程包括两个独立的步骤：生成通过激光解吸，并通过ESI样品颗粒的后续后电离气体样品分子。 ELDI生成多电荷离子，这是适合于自上而下蛋白质鉴定与各种MS / MS技术（Peng等人，“电辅助激光解吸电离（ELDI）和肽的串联质谱法和蛋白质”。快速COMMUN质谱Spectrom 2007; 21：2541年至2546年）。 ELDI被示出为能够表征大的蛋白质达29千道尔顿。由ELDI机构产生多电荷蛋白可以通过降低pH值，或通过在溶液ESI加入m-硝基苄醇（M-NBA）转移到更高的电荷状态。无ELDI允许在ELDI过程中发生的化学反应。反应ELDI实验证明是有效的气相形成核糖核酸酶A（RNA酶A）-CTP复杂，并且为上线用二硫苏糖醇（DTT）二硫化物还原为氧化型谷胱甘肽和胰岛素。这些数据提供了直接的蛋白 - 配体结合研究的可能性，以及上线自上而下蛋白质鉴定为蛋白质组学研究中，不费力的样品制备的必要性。

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