Analytical properties of the nanoelectrospray ion source
Analytical properties of the nanoelectrospray ion source
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DOI：
10.1021/ac9509519
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发表时间：
1996-01-01
影响因子：
7.4
通讯作者：
Mann, M
中科院分区：
文献类型：
--
作者：
Wilm, M;Mann, M
纳米电喷雾离子源（nanoES）最近被开发出来并进行了理论描述。它与传统的电喷雾源和其它小型化电喷雾源的不同之处在于：（i）通过将喷雾毛细管拉至细尖端而实现其1-2 μ m的喷雾孔，（ii）其类似于20 nL/min的非常低的流速和其产生的小尺寸液滴，以及（iii）没有溶剂泵和入口阀。详细描述了nanoES针的制造和操作。含盐量高达0.1 M的溶液可以在没有护套或气动辅助的情况下喷涂。nanoES中改进的去溶剂化导致糖蛋白信号的仪器限制分辨率，并且信号平均的能力广泛地允许40 kDa蛋白质的C-末端测序。通过验证93%的碳酸酐酶序列，证明了对未分离的肽混合物的组分进行了广泛的质谱和串联质谱研究。与nanoES程序相结合的快速和稳健的洗脱/浓缩步骤允许直接分析不纯样品，例如凝胶内消化后提取的肽混合物。
The nanoelectrospray ion source (nanoES) has recently been developed and described theoretically. It is different from conventional electrospray sources and from other miniaturized electrospray sources by (i) its 1-2 mu m spraying orifice achieved by pulling the spraying capillary to a fine tip, (ii) its very low now rate of similar to 20 nL/min and the small size of droplets it generates, and (iii) the absence of solvent pumps and inlet valves. The fabrication and operation of nanoES needles is described in detail. Solutions with up to 0.1 M salt contents could be sprayed without sheath now or pneumatic assist. Improved desolvation in nanoES led to instrument-limited resolution of the signals of a glycoprotein and the ability to signal average extensively allowed the C-terminal sequencing of a 40 kDa protein. Extensive mass spectrometric and tandem mass spectrometric investigation of the components of an unseparated peptide mixture was demonstrated by verification of 93% of the sequence of carbonic anhydrase. A rapid and robust desalting/concentration step coupled to the nanoES procedure allows the direct analysis of impure samples such as peptide mixtures extracted after in-gel digestion.