福田 宏之 日本パーセプティブ(株), 技術開発課, (研究職)課長
IMAMOTO Naoko Faculty of Medicine., Osaka Univ., Professor, Instructor, 医学部, 助手 (20202145)
SHIMONISHI Yasutsugu Inst.For Protein Res., Osaka Univ., Professor, 蛋白質研究所, 教授 (00029951)
FUKUDA Hiroyuki Nihon PerSeptive Ltd., R&D,Manager
|Budget Amount *help
¥6,200,000 (Direct Cost : ¥6,200,000)
Fiscal Year 1997 : ¥1,200,000 (Direct Cost : ¥1,200,000)
Fiscal Year 1996 : ¥5,000,000 (Direct Cost : ¥5,000,000)
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.