不同理化因素对重组人成纤维细胞生长因子21蛋白纯度的影响

doi:10.13481/j.1671-587x.20150314

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (03): 506-509.doi: 10.13481/j.1671-587x.20150314

不同理化因素对重组人成纤维细胞生长因子21蛋白纯度的影响

王长文¹, 刘敏¹, 刘磊², 姜勇², 张磊², 李艳², 王会岩²

1. 吉林医药学院公共卫生学院营养与食品卫生学教研室, 吉林吉林 132013;
2. 吉林医药学院检验学院生物技术教研室, 吉林吉林 132013

收稿日期:2014-09-02 发布日期:2015-08-01
通讯作者: 王会岩,副教授(Tel:0432-64560324,E-mail:zswhy518@163.com) E-mail:zswhy518@163.com
作者简介:王长文(1966-),男,吉林省松原市人,副教授,农学博士,主要从事食品营养与食品卫生方面的研究。
基金资助:
国家自然科学基金资助课题(81273421);吉林省科技厅科技发展计划项目资助课题(20116044);吉林省教育厅科研基金资助课题(361号)

Effects of different physical and chemical factors on purity of recombinant human fibroblast growth factor 21

WANG Changwen¹, LIU Min¹, LIU Lei², JIANG Yong², ZHANG Lei², LI Yan², WANG Huiyan²

1. Department of Nutrition and Food Hygiene, School of Public Health, Jilin Medical College, Jilin132013, China;
2. Department of Biotechnology, School of Medical Laboratory, Jilin Medical College, Jilin 132013, China

Received:2014-09-02 Published:2015-08-01

摘要/Abstract

摘要：

目的:研究重组人成纤维细胞生长因子21(rhFGF21)在不同保存条件下的稳定性,阐明温度、pH值、缓冲溶液和反复冻融因素对rhFGF21稳定性的影响,进一步完善纯化rhFGF21的工艺。方法:rhFGF21在不同温度(-20℃、4℃和25℃)、不同pH值缓冲液(醋酸-醋酸钠缓冲液pH 4.0和pH 5.0、柠檬酸-柠檬酸钠缓冲液pH 5.0和pH 6.0、磷酸氢二钠-磷酸二氢钠缓冲液pH 6.0、pH 6.5和pH 7.0,Tris-HCl缓冲液pH 7.5和pH 8.0)中及反复冻融条件下分别保存,采用高效液相色谱法(HPLC)检测rhFGF21纯度,同时观察外观性状。结果:于-20℃和4℃分别保存时,rhFGF21的各项性质在观察期内均无明显变化;25℃时,rhFGF21存放14 d纯度由0 d时(100.00±0.00)% 下降至(15.20±0.14)% (P<0.01);在pH4.0和pH 5.0缓冲液中25℃条件下保存7 d,溶液出现浑浊;在pH 5.0和pH 6.0缓冲液中25℃条件下保存7 d,溶液出现浑浊;在pH 6.5和pH 8.0 磷酸盐和Tris-HCl缓冲液中,溶液澄清透明;rhFGF21原液反复冻融后其纯度变化差异有统计学意义(P<0.05)。结论:rhFGF21在低温和pH 6.5~8.0磷酸盐和Tris-HCl缓冲液中溶液较稳定,反复冻融处理对rhFGF21稳定性也有明显影响。

关键词: 重组人成纤维细胞生长因子21, 纯度, 稳定性, 理化因素

Abstract:

Objective To study the stabilities of recombinant human fibroblast growth factor 21 (rhFGF21) deposited at different conditions, and to elucidate the influence of temperature, buffer solutions and repeated freeze thawing in the stability of rhFGF21. Methods rhFGF21 was deposited at different conditions such as temperatures (-20℃, 4℃, 25℃), buffer solutions (acetate sodium acetate buffer pH 4.0 and pH 5.0, citric acid-sodium citrate buffer pH 5.0 and pH 6.0, disodium hydrogen phosphate and sodium dihydrogen phosphate buffer pH 6.0, pH 6.5and pH 7.0, Tris - HCl buffer pH 7.5 and pH8.0) and repeated freeze thawing, respectively.The purity of rhFGF21 was determined by HPLC, and the appearance was also observed. Results All properties of rhFGF21 were not altered during the observation period at -20℃ and 4℃;the purity of rhFGF21 deposied for 14 d was reduced to (15.20±0.14)% compared with 0 d(100%±0.00%) at 25℃(P<0.05).The acetate sodium acetate buffer pH 4.0 and pH 5.0 and citric acid sodium citrate buffer pH 5.0 and pH 6.0 were turbidity under the condition of 25℃ for 7 d;the disodium hydrogen phosphate and sodium dihydrogen phosphate buffer pH 6.5 and pH 7.0, and Tris-HCl buffer pH 7.5 and pH 8.0 were clear transparent under the condition of 25℃ for 7 d. The purity of rhFGF21 had significant change after repeated with freeze thawing (P<0.05). Conclusion rhFGF21 is stable when deposited at low temperature and pH 6.5-8.0 neutral conditions.High temperature, acidic and basic conditions are harmful for the stability of rhFGF21.It is profit for rhFGF21 stability in sodium dihydrogen phosphate buffer.There is impact on rhFGF21 stability after freeze thrawing repeatedly.

Key words: recombinant human fibroblast growth factor 21, purity, stability, physical and chemical factors

中图分类号:

王长文, 刘敏, 刘磊, 姜勇, 张磊, 李艳, 王会岩. 不同理化因素对重组人成纤维细胞生长因子21蛋白纯度的影响[J]. 吉林大学学报(医学版), 2015, 41(03): 506-509.

WANG Changwen, LIU Min, LIU Lei, JIANG Yong, ZHANG Lei, LI Yan, WANG Huiyan. Effects of different physical and chemical factors on purity of recombinant human fibroblast growth factor 21[J]. Journal of Jilin University Medicine Edition, 2015, 41(03): 506-509.

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不同理化因素对重组人成纤维细胞生长因子21蛋白纯度的影响

Effects of different physical and chemical factors on purity of recombinant human fibroblast growth factor 21

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