异硫氰酸荧光素标记尿激酶的制备与性能研究  

Preparation and Property of Fluorescein Labeled Urokinase

作　　者：张雅娟[1] 俞洁[1] 张冬未 梁洪泽[1] 梁明明[2] 赵玲玲[1] 

机构地区：[1]宁波大学材料科学与化学工程学院,浙江宁波315211 [2]马鞍山市人民医院,安徽马鞍山243000

出　　处：《宁波大学学报（理工版）》2016年第3期78-82,共5页Journal of Ningbo University：Natural Science and Engineering Edition

基　　金：国家自然科学基金(51403108);浙江省自然科学基金(LQ13B040001);北京分子科学国家实验室开放基金;宁波市自然科学基金(2013A610091);宁波大学学科基金项目(XYL14013;XKZSC05);宁波大学王宽诚幸福基金

摘　　要：用异硫氰酸荧光素(FITC)对溶栓蛋白药物尿激酶(uPA)进行了荧光标记,制备了荧光素标记的尿激酶(FITC-u PA),经核磁谱及红外谱表征证实了反应能有效进行,其荧光素/蛋白值(F/P)值为0.45.尿激酶经荧光素标记后,具有良好的荧光性质,其荧光检测下限低达10-6 g?m L^(-1).在1~100mg?m L^(-1)范围内,荧光强度与溶液浓度具有很好的线性关系,可用于微/痕量蛋白质的定量/定性检测和跟踪标记.荧光素标记后的尿激酶,其流体力学直径与未标记的尿激酶基本一致,其体外溶栓能力也与未标记的尿激酶相当,说明FITC标记基本不影响尿激酶的生物活性,是一种简单、有效的溶栓药物标记方法.Fluorescein labeled urokinase （FITC-uPA） is synthesized by the reaction of fluorescein isothiocyanate （FITC） with urokinase （uPA）, a kind of protein drug for thrombolysis. The reaction is confirmed by1HNMR and FT-IR. The ratio （F/P） of fluorescein over protein is 0.45. The urokinase is found to possess good fluorescent property after fluorescein is labeled, and the detection limit of fluorescence is as low as 10-6 g·mL-1. There is obvious linear relationship between the fluorescein intensity and concentration of FITC-uPA within the concentration range of 1-100μg-mL-1, which can be used for qualitative/quantitative detection and tracking of micro/trace proteins. FITC-uPA has similar hydrodynamic diameter and thrombolysis efficiency with the unlabeled urokinase. Based on the study, it can be concluded that the fluorescein-label is a simple and effective method for thrombolysis drug labeling without affecting the biological active of urokinase.

关 键 词：异硫氰酸荧光素 荧光标记 尿激酶 溶栓 

分 类 号：TQ937[化学工程]