Characterization of Prolidase Activity Using Capillary Electrophoresis with Tris(2,2'-bipyridyl)ruthenium(II) Electrochemiluminescence Detection and Application To Evaluate Collagen Degradation in Dia

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• 作者：Jipei Yuan ; Tao Li ; Xue-Bo Yin ; Li Guo ; Xiuzhen Jiang ; Wenrui Jin ; Xiurong Yang ; and Erkang Wang
• 刊名：Analytical Chemistry
• 出版年：2006
• 出版时间：May 1, 2006
• 年：2006
• 卷：78
• 期：9
• 页码：2934 - 2938
• 全文大小：89K
• 年卷期：v.78,no.9(May 1, 2006)
• ISSN：1520-6882

文摘

A new method for prolidase (PLD, EC 3.4.13.9) activityassay was developed based on the determination ofproline produced from enzymatic reaction through capillary electrophoresis (CE) with tris(2,2'-bipyridyl)ruthenium(II) [Ru(bpy)₃²⁺] electrochemiluminescence detection (ECL). A detection limit of 12.2 fmol (S/N = 3) forproline, corresponding to 1.22 × 10^-8 units of prolidasecatalyzing for 1min was achieved. PLD activity determinedby CE-ECL method was in agreement with that obtainedfrom the classical Chinard's one. CE-ECL showed itspowerful resolving ability and selectivity as no samplepretreatment was needed and no interference existed. Theclinical utility of this method was successfully demonstrated by its application to assay PLD activity in theserum of diabetic patients in order to evaluate collagendegradation in diabetes mellitus (DM). The results indicated that enhanced collagen degradation occurred in DM.