Development of peptide array for high sensitive detection of phosphorylation signal using MALDI-MS

研究了一种使用氧化钛板的肽阵列用于检测蛋白激酶活性。 由于其具有去除自身表面非特异性吸附的自清洁能力，并在MALDI质谱中起到无机基质的作用，因此有望实现更准确、高灵敏度的蛋白激酶活性分析。 首先，采用溶胶-凝胶法制备了氧化钛平板。 在紫外光照射下，它表现出光诱导的亲水性，并且其表面上的非特异性吸附仅通过用水洗涤就被去除。 接下来，通过配备有MALDI质谱仪（MS）的激光器，使用3-氨基丙基三乙氧基硅烷和光裂解的接头将底物肽固定在表面上。 最后，对所制备的肽阵列进行了质谱分析，为进一步开发基于氧化钛板的高灵敏度MALDI-MS检测肽阵列奠定了基础。

A peptide array using a titanium oxide plate has been investigated for detection of protein kinases activities. Because the one has self-cleaning ability to remove non-specific adsorption on the surface of itself, and plays a role of inorganic matrix at MALDI mass spectrometry, it will be expected to be able to realize more accurate analysis of protein kinases activities with high sensitivity. First, the titanium oxide plate was prepared by sol-gel method. It showed photo-induced hydrophilicity at UV irradiation and much of non-specific adsorption on its surface was removed only by washing with water. Next, substrate peptides were immobilized on the surface using 3-aminopropyl triethoxysilane and photo-cleaved linker by laser equipped with a MALDI mass spectrometer (MS). Finally, the peptide array were analyzed by MS. From above results, we would establish the basis technique to develop the peptide array using the titanium oxide plate for high sensitive detection at MALDI-MS.