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

蛋白质一级结构微量分析全自动气相化学反应装置的研制

基本信息

批准号：
08558071
负责人：
TAKAO Toshifumi
金额：
$ 3.97万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08558071/
关键词：
Microanalysis Anticorrosive system Gas-phase chemical reaction Protein primary structure Specific chemical cleavage Gas-assisted flow machinery Mass spectrometry

项目摘要

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.

设备。它可以用于与基质辅助激光解吸质谱法（MALDI-TOFMS）结合使用肽或蛋白质原代结构的微分析。该设备的出色特征是：1）可用于任何气相化学反应，避免样品丢失和污染。可以优化反应以有效地产生产品，2）样品板（46x48mm），该样品板（46x48mm）用金色镀金，并用Teflon将60个样品固定在最大的样品中，并且可以将60个样品固定在两个设备和设备的构造中，并构成了一个设备和样品室的设备。 That, all the samples on the plate are subjected to chemical reactions in gaseous phase at once.The equipment consists of : 1) a solution 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两阶段真空系统，可以有效地去除腔室中的试剂，4）个人计算机，该计算机可以根据反应的时间计算机自动控制腔室内的电动阀和温度。该设备已经检查了几种化学反应。结果，已修改以实现的Edman降解，并且可以在Picomole水平的Picomole水平或蛋白质上进行蛋氨酸特异性的裂解，其产物有效地由Maldi-Tofms测量。