中文摘要：

发展了一种基于毛细管电泳（CE）-激光诱导荧光（LIF）检测的多个细胞内源激酶的抑制剂平行筛选及选择性评价方法。CE高效的分离能力和LIF检测 器的高选择性，使得同时测试多个胞内激酶的活性成为可能。共4种细胞系、3种特异性蛋白激酶底物肽、2种选择性蛋白激酶抑制剂和1种非选择性蛋白激酶抑制利用于方法的建移。特异性底物肽与细胞裂解液混合后孵育，被其相应的激酶选择件地磷酸化，利用CE-LIF分离检测磷酸化产物和底物肽，同时测定一个抑制剂对几种蛋白激酶的抑制活性，用于评价抑制剂的选择性。与传统的坼靶机筛选帙式相比，这种基于细胞裂解液的多靶标筛选方法能提供更多的信息，更加高效，且细胞裂解液作为一种廉价的激酶来源大大降低了9带选成本。

英文摘要：

A method that can be used for screening protein kinase inhibitors （PKIs） and simultaneously assess- ing their selectivity is described. The method is based on simultaneously assaying multiple cellular protein kina- ses by performing capillary electrophoresis （CE） separation and measuring the peak areas of tile phosphoryla- ted substrate peptides. The powerful separation capability of CE combined with the highly sensitive and selec- tive laser-induced fluorescence （LIF） detector enables the direct screening of PKIs against cell lysaies, which are used as an inexpensive source of enzymes. Four cell lines, three specific substrate peptides labeled with 5- carboxyfluorescein （5-FAM） , two relative specific PKIs （ TBB and H-89 ） and one non-specific PKI （ staurospo- rine ） were utilized to prove the methodology. With this method, the inhibitory activity of the tested compounds against multiple protein kinases was identified in parallel by comparing the peak areas of the phosphorylated substrates with those obtained in the absence of any inhibitors. The reduced peak area of the phosphorylated substrate definitively represents a positive screening result. Simultaneously, assaying the inhibition of one inhibitor against mutiple cellular protein kinases enables the assessment of its selectivity. Compared to the con- ventional, single-target screening format, the cell lysate-based multi-target method is more informative, more straightforward and more cost effective.