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

使用氧化钛板的肽阵列已被研究用于检测蛋白激酶的活性。由于它具有自清洁能力，可以去除自身表面的非特异性吸附，并且在MALDI质谱中扮演无机基质的角色，因此有望实现对蛋白激酶活性的更准确、高灵敏度的分析。首先，采用溶胶-凝胶法制备氧化钛板。在紫外线照射下表现出光诱导的亲水性，仅用水清洗即可去除表面的大部分非特异性吸附。利用MALDI质谱仪（MS），采用3-氨基丙基三乙氧基硅烷和光裂解连接剂将底物肽固定在表面。最后，利用质谱法对多肽阵列进行分析，为利用氧化钛板构建多肽阵列进行MALDI-MS高灵敏度检测奠定了基础技术。

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.