人淋巴细胞犬尿氨酸酶活性的测定  

Determination of Activity of Human Lymphocyte Kynureninase

作　　者：沈杰[1] 高平进[1] 顾天华[1] 朱鼎良[1] 

机构地区：[1]上海交通大学医学院瑞金医院上海市高血压研究所,上海200025

出　　处：《上海交通大学学报（医学版）》2006年第11期1239-1241,共3页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(30270545)资助项目

摘　　要：目的 建立人淋巴细胞犬尿氨酸酶活性的高效液相色谱检测法。方法 以人淋巴细胞总蛋白作为酶的来源，以3-羟基犬尿氨酸为底物，5’磷酸吡哆醛为辅酶，进行酶促反应，产物经ODS色谱柱分离后用荧光检测器定量检测，从单位时间内产物的增加量来计算犬尿氨酸酶的催化活性。结果 该方法检测产物的线性范围是2～400nmol／L。方法的重复性好，日内变异系数（CV）≤3．0％，日间CV≤3．2％。结论 该方法适用于测定各种生物组织样本中犬尿氨酸酶的活性。Objective To develop an efficient method for the determination of activity of human lymphocyte kynureninase by high performance liquid chromatography. Methods Protein containing kynureninase was extracted from lymphocytes. The reaction was made with 3-hydroxyanthranilic acid as suhstrate and pyridoxal-5＇-phosphate as coenzyme. The product was determined by high performance liquid chromatography and fluorescence detection. Results Standard curve of 3-hydroxyanthranilic acid was highly linear over the range from 2 to 400 nmol/L. The intra-day coefficient of variation（CV） was less than 3.0% and the inter-day CV was less than 3.2%. Conclusion The developed assay is efficient and can he used to determine the activity of kynureninase from kinds of tissues.

关 键 词：犬尿氨酸酶 淋巴细胞 活性 高效液相色谱法 

分 类 号：R446.1[医药卫生—诊断学]