Development of a Full-Automatic Equipment for Gas-Phase Chemical Reaction Intended for Microanalysis of Protein Primary Structures

研制了一种与基质辅助激光解吸质谱联用的多肽或蛋白质一级结构微量分析装置。该设备的突出特点是：1）可用于任何气相化学反应，避免样品损失和污染，可优化反应以有效地产生产物; 2）样品板（46 × 48 mm），镀金，单面涂有聚四氟乙烯;最多可容纳60个样品，并被设置在设备的反应室和质谱仪的样品板保持器中。该装置由以下部分组成：1）溶剂或试剂输送系统，其各处均为防腐的，包括六个试剂瓶、输送管线、电动阀和气体辅助流动机械; 2）密封的玻璃器皿反应室（体积：180 cm^3）与控制高达110 ° C的加热块结合，3）两级真空系统，通过该系统可以有效地去除腔室中的试剂，4）个人电脑，其可根据反应的时间程序自动控制电动阀和室内温度。已经用这种设备研究了几种化学反应。结果，Edman降解，这已被修改为实现相当简单，和甲硫氨酸特异性切割与溴化氰可以进行皮摩尔水平的肽或蛋白质，其产品有效地测量MALDI-TOFMS。

An equipment.which can be applied for microanalysis of peptide or protein primary structures in conjunction with matrix-assisted laser desorption mass spectrometry (MALDI-TOFMS), has been developed. The outstanding features of the equipment are : 1) It can be used for any gas-phase chemical rections, which avoid sample loss and contamination.The reaction may be optimized to efficiently yield the products, 2) A sample plate (46x48mm), which is gilded with gold and one-sidedly coated with Teflon, can hold 60 samples at maximum and is set into both the reaction chamber of the equipment and a sample-plate holder for a mass spectrometer. Thus, all the samples on the plate are subjected to chemical reactions in gaseous phase at once.The equipment consists of : 1) a solvent or reagent delivery system, everywhere of which is anticorrosive and involves six reagent bottles, delivery lines, electric valves, and gas-assisted flow machinery, 2) an airtight glassware reaction chamber (volume : 180 cm^3) in conjunction with a heating block controlled up to 110ﾟC,3) a two-stage vacuum system, by which the reagent in the chamber can be efficiently removed, 4) a personal computer, which can automatically control the electric valves and temperature inside the chamber according to a time-program of a reaction. Several chemical reactions have been examined with this equipment. As a result, Edman degradation, which has been modified to be achieved quite simply, and methionine-specific cleavage with cyanogen bromide could be carried out on picomole level of peptides or proteins, the products of which were effectively measured by MALDI-TOFMS.