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

研制了一种可与基质辅助激光解吸质谱仪(MALDI-TOFMS)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)它可以用于任何气相化学分离，避免了样品损失和污染。反应可以进行优化，以高效地产生产品。2)一个镀金并单面涂有特氟龙的样品板(46x48 Mm)最多可以容纳60个样品，并被放置在设备的反应室和质谱仪的样品板支架中。该设备包括：1)一个防腐的溶剂或试剂输送系统，其中包括6个试剂瓶、输送管道、电动阀门和气辅流动机械；2)密闭玻璃器皿反应室(体积：180 cm^3)与控制高达110ﾟC的加热块；3)两级真空系统，可有效地去除室内试剂；4)个人计算机，可根据反应时间程序自动控制电动阀门和室内温度。用这台设备已经研究了几个化学反应。因此，可以在多肽或蛋白质的皮摩尔水平上进行Edman降解，其产物可以用MALDI-TOFMS有效地测定。

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.