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

一个设备。它可以与基质辅助激光解吸质谱（MALDI-TOFMS）结合应用于肽或蛋白质初级结构的微分析。该设备的突出特点是：1)可用于任何气相化学反应，避免了样品损失和污染。2)样品板（46x48mm），表面镀金，一面涂有聚四氟乙烯，最大可容纳60个样品，设置在设备的反应室和质谱仪的样品板支架上。因此，板上的所有样品都在气相中同时发生化学反应。该设备包括：1)溶剂或试剂输送系统，其各处都是防腐的，包括六个试剂瓶，输送管线，电动阀和气体辅助流动机械；2)密封玻璃器皿反应室(体积：180 cm^3)配合加热块控制高达110ºC，3)两级真空系统，通过该系统可以有效地去除腔内的试剂，4)个人电脑，可以根据反应的时间程序自动控制腔内的电动阀和温度。用这台设备检测了几种化学反应。利用MALDI-TOFMS，可以在微分子水平上对多肽或蛋白质进行Edman降解和与溴化氰的蛋氨酸特异性裂解。

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.