NV蛋白酶冻干工艺的相关技术参数

doi:10.13481/j.1671-587x.20170136

吉林大学学报(医学版) ›› 2017, Vol. 43 ›› Issue (01): 181-185.doi: 10.13481/j.1671-587x.20170136

NV蛋白酶冻干工艺的相关技术参数

张连芝¹, 洪敏², 程熠³

1. 吉林大学基础医学院生物化学与分子生物学实验教学中心, 吉林长春 130021;
2. 吉林大学基础医学院生物化学与分子生物学系, 吉林长春 130021;
3. 吉林大学第一医院转化医学研究院, 吉林长春 130061

收稿日期:2016-08-10 出版日期:2017-01-28 发布日期:2017-02-08
通讯作者: 程熠,副教授,硕士研究生导师(Tel:0431-88782222,E-mail:chengyi@jlu.edu.cn) E-mail:chengyi@jlu.edu.cn
作者简介:张连芝(1964-),女,吉林省长春市人,理学硕士,主要从事重组蛋白制备及应用方面的研究。
基金资助:
吉林省卫计委科研项目资助课题（2008Z043）；吉林省科技厅国际合作项目资助课题（20140413016GH）；吉林省科技厅自然科学基金资助课题（20150101130JC）

Relative parameters of lyophilization of NV protease

ZHANG Lianzhi¹, HONG Min², CHENG Yi³

1. Experimental Center of Biochemistry and Molecular Biology, Jilin University, Changchun 130021, China;
2. Department of Biochemistry and Molecular Biology, Jilin University, Changchun 130021, China;
3. Institute of Translational Medicine, First Hospital, Jilin University, Changchun 130061, China

Received:2016-08-10 Online:2017-01-28 Published:2017-02-08

摘要/Abstract

摘要：

目的：研究NV蛋白酶冻干流程，为冻干工艺的确定提供相关技术参数。方法：制备NV蛋白酶溶液，测定不同温度、不同pH值条件下NV蛋白酶的圆二色谱；以偶氮酪蛋白方法检测NV蛋白酶的活性；采用双阶段冻干法冻干NV蛋白酶；测定Tween 80作为NV蛋白酶冻干变性防护剂的权宜浓度和权宜冷冻速率。结果：NV蛋白酶复溶活性和溶解度随平均冷冻速率升高而降低。Tween 80作为NV蛋白酶冻干变性防护剂平衡NV蛋白酶复溶活性和溶解度的权宜浓度为0.01%。降低冻干药品生产时间成本的权宜冷冻速率参数为2.0℃·min^-1。结论：NV蛋白酶具有一定的抵抗冻干过程中温度和pH值降低的能力，平均冷冻速率影响NV蛋白酶复溶活性和溶解度；0.01% Tween 80和2.0℃·min^-1平均冷冻速率是NV蛋白酶生产性冻干的有效技术选择。

关键词: 蛋白酶, 冷冻速率, 冻干, 冻干保护剂

Abstract:

Objective: To investigate the lyophilization process of NV protease,and to provide the relative parameters of lyophilization of NV protease.Methods: The NV protease solution was prepared. The circular dichroism of NV protease was analyzed under the condition of different temperatures and pH values. The activity of NV protease was measured using azocasein. The NV protease was lyophilized using two-step lyophilization. The concentration of expediency and the freezing rate of expediency of Tween 80 as a cryoprotectant of NV protease were detected.Results: The activity and solubility of lyophilized NV protease were decreased with the increase of average freezing rate. The expedient of Tween 80 was 0.01% in balancing the activity and solubility of lyophilized NV protease, and the optimal average freezing rate was 2.0℃·min^-1 to reduce the time cost of lyophilization.Conclusion: NV protease is resistant to the reduction of the temperature and pH value during lyophilization. The average freezing rate affects the activity and solubility of lyophilized NV protease. The combination of 0.01% Tween 80 and 2.0℃·min^-1 average freezing rate is an effective choice for NV protease lyophilization.

Key words: lyophilization, protease, cryoprotectant, freezing rate

中图分类号:

张连芝, 洪敏, 程熠. NV蛋白酶冻干工艺的相关技术参数[J]. 吉林大学学报(医学版), 2017, 43(01): 181-185.

ZHANG Lianzhi, HONG Min, CHENG Yi. Relative parameters of lyophilization of NV protease[J]. Journal of Jilin University Medicine Edition, 2017, 43(01): 181-185.

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NV蛋白酶冻干工艺的相关技术参数

Relative parameters of lyophilization of NV protease

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