Single-neuron analysis using CE combined with MALDI MS and radionuclide detection
Single-neuron analysis using CE combined with MALDI MS and radionuclide detection
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DOI：
10.1021/ac0156621
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发表时间：
2002-02-01
影响因子：
7.4
通讯作者：
Sweedler, JV
中科院分区：
文献类型：
--
作者：
Page, JS;Rubakhin, SS;Sweedler, JV
毛细管电泳 (CE) 与基质辅助激光解吸/电离质谱 (MALDI MS) 和放射性核素检测相结合，可分析质量有限的生物样品。 Nanovial 采样技术能够将 50 至 150 nL 体积的样品注射到 CE 毛细管中；分离后，纳升级分收集将 CE 流出物与 MALDI 基质结合在一起，并最大限度地减少样品扩散，从而允许对 CE 级分进行 MALDI MS 和放射性核素检测。 MAIM MS 通过提供准确的分子质量数据补充了 CE 的洗脱时间信息，放射性核素检测提供了 zeptomole 检测限和定量信息。虽然 MALDI MS 可检测足够浓度的所有完全加工的肽，但在含有 S-35-Met 的培养基中培养神经元可对新合成的含蛋氨酸肽进行选择性放射性核素检测。通过 CE/MALDI MS/放射性核素检测对加州海兔的单个 40-mum 袋细胞神经元中预期的神经肽和激素进行分析和检测，证明了这种联用方法处理化学复杂的质量限制样品的能力。
Capillary electrophoresis (CE) has been combined with matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) and radionuclide detection to assay mass-limited biological samples. Nanovial sampling techniques enable injections into the CE capillary from 50 to 150-nL volume samples; after the separation, nanoliter fraction collection combines the CE effluent with a MALDI matrix and minimizes sample spreading, thus allowing both MALDI MS and radionuclide detection on the CE fractions. MAIM MS complements the elution time information of CE by providing accurate molecular mass data, and radionuclide detection provides zeptomole limits of detection with quantitative information. While MALDI MS detects all fully processed peptides at sufficient concentration, culturing the neuron in media containing S-35-Met provides selective radionuclide detection of newly synthesized methionine-containing peptides. The analysis and detection of the expected neuropeptides and hormones in a single 40-mum bag cell neuron from Aplysia californica with CE/MALDI MS/radionuclide detection demonstrates the ability of this hyphenated approach to work with chemically complex mass-limited samples.