异硫氰酸荧光素标记尿激酶的制备与性能研究
PDF下载 (375)张雅娟1,俞 洁1,张冬未1,梁洪泽1,梁明明2,赵玲玲1*.异硫氰酸荧光素标记尿激酶的制备与性能研究[J].宁波大学学报(理工版),2016,29(3):78-82.DOI:
ZHANG Ya-juan1,YU Jie1,ZHANG Dong-wei1,LIANG Hong-ze1,LIANG Ming-ming2,ZHAO Ling-ling1*.Preparation and Property of Fluorescein Labeled Urokinase[J].Journal of Ningbo University(Natural Science & Engineering Edition),2016,29(3):78-82.DOI:
| Title: | Preparation and Property of Fluorescein Labeled Urokinase |
| 作者: | 张雅娟1, 俞 洁1, 张冬未1, 梁洪泽1, 梁明明2, 赵玲玲1* |
| Author(s): | ZHANG Ya-juan1, YU Jie1, ZHANG Dong-wei1, LIANG Hong-ze1, LIANG Ming-ming2, ZHAO Ling-ling1* |
| 关键词: | 异硫氰酸荧光素; 荧光标记; 尿激酶; 溶栓 |
| Keywords: | fluorescein isothiocyanate; fluorescein-label; urokinase; thrombolysis |
| 分类号: | TQ937 |
| 文献标识码: | A |
| 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-uPA), 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?mL-1. 在1~100 ?g?mL-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. |
| Abstract: | 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 by 1HNMR 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 1-100 μg?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. |
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| 备注/Memo: | 收稿日期: 2014?12?9. 宁波大学学报（理工版）网址: http://journallg.nbu.edu.cn/ 基金项目: 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. 第一作者: 张雅娟（1989－）, 女, 河北唐山人, 在读硕士研究生, 主要研究方向: 生物医用材料. E-mail: zyjsimple@163.com *通信作者: 赵玲玲（1986－）, 女, 湖南怀化人, 讲师, 主要研究方向: 生物医用材料. E-mail: zhaolingling@nbu.edu.cn 宁波大学学报（理工版）网址：http://journallg.nbu.edu.cn/ |
