NV蛋白酶冻干工艺的相关技术参数

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

Abstract

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

CLC Number:

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.

References

[1] 程熠,史方亮,王之光,等.溶栓素的纯化研究[J].白求恩医科大学学报,2001,27(1):9-11.
[2] 程熠,洪敏,李奇,等.阴、阳离子交换示差组合色谱法的设计及在纤溶酶纯化中的应用[J]. 吉林大学学报:医学版,2005,31(5):699-701.
[3] 程熠,洪敏,张云龙. 溶栓素产品纯度的检测[J]. 吉林大学学报:医学版,2004,30(5):816-818.
[4] 程熠,洪敏,张云龙. 溶栓素定量方法的筛选系统[J]. 吉林大学学报:医学版,2004,30(6):981-983.
[5] 李奇,洪敏,付海英. 溶栓素抗凝血和抗血栓形成作用的实验研究[J]. 吉林大学学报:医学版,2004,30(3):401-404.
[6] 李奇,迟秀梅,石立红,等. N-V蛋白酶体内外纤维蛋白溶解活性的实验研究[J]. 中国药理学通报,2006,22(8):976-979.
[7] Peters BH, Staels L, Rantanen J, et al. Effects of cooling rate in microscale and pilot scale freeze-drying-variations in excipient polymorphs and protein secondary structure[J]. Eur J Pharm Sci,2016, 95:72-81.
[8] Ulrichs T, Drotleff AM, Ternes W. Determination of heat-induced changes in the protein secondary structure of reconstituted livetins (water-soluble proteins from hen's egg yolk) by FTIR[J]. Food Chem, 2015, 172:909-920.
[9] Park J, Nagapudi K, Vergara C, et al. Effect of pH and excipients on structure, dynamics, and long-term stability of a model IgG1 monoclonal antibody upon freeze-drying[J]. Pharm Res,2013, 30(4):968-984.
[10] Depaz RA, Pansare S, Patel SM. Freeze-drying above the glass transition temperature in amorphous protein formulations while maintaining product quality and improving process efficiency[J]. J Pharm Sci, 2016, 105(1):40-49.
[11] Chang BS, Kendrick BS, Carpenter JF. Surface-induced denaturation of proteins during freezing and its inhibition by surfactants[J]. J Pharm Sci, 1996, 85(12):1325-1330.
[12] Sarciaux JM, Hageman MJ. Effects of bovine somatotropin (rbSt) concentration at different moisture levels on the physical stability of sucrose in freeze-dried rbSt/sucrose mixtures[J]. J Pharm Sci, 1997, 86(3):365-371.
[13] Strambini GB, Gabellieri E. Proteins in frozen solutions:evidence of ice-induced partial unfolding[J]. Biophys J, 1996, 70(2):971-976.
[14] Jiang S, Nail SL. Effect of process conditions on recovery of protein activity after freezing and freeze-drying[J]. Eur J Pharm Biopharm, 1998, 45(3):249-257.
[15] Cleland JL, Powell MF, Shire SJ. The development of stable protein formulations:a close look at protein aggregation, deamidation, and oxidation[J]. Crit Rev Ther Drug Carrier Syst, 1993, 10(4):307-377.
[16] Knepp VM, Whatley JL, Muchnik A, et al. Identification of antioxidants for prevention of peroxide-mediated oxidation of recombinant human ciliary neurotrophic factor and recombinant human nerve growth factor[J]. PDA J Pharm Sci Technol, 1996, 50(3):163-171.

Relative parameters of lyophilization of NV protease

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 6

Metrics

Comments

Recommended 0

[1]	CHEN Chunyu, LIU He, HE Pan, LU Jianzhong, YING Di, MA Ning. Inhibitory effect of wild chrysanthemum extract on activity of trypsin-like protease produced by Porphyromonas gingivalis [J]. Journal of Jilin University Medicine Edition, 2015, 41(05): 941-945.
[2]	LI Yunqian, ZHAO Liyan, SHI Yan, LIU Hui, MIAO Chunsheng, XU Wanzhen, YANG Zhiqing. Influence of overexpression of miR-181b in expressions of MT1-MMP and TIMP3 protein and invasive ability in glioma U87 cells [J]. Journal of Jilin University Medicine Edition, 2015, 41(05): 925-931.
[3]	ZHU Yang, LI Daowei, FANG Tengjiaozi, SHI Ce, WANG Dandan, SUN Hongchen. Promotive effect of PLGA nanofiber scaffold adsorped Epo gene mediated by adenovirus on bone defect repair [J]. Journal of Jilin University Medicine Edition, 2015, 41(04): 711-715.
[4]	QI Ling, WANG Weiyao, ZHANG Yanchun, CAI Shangxia, YANG Ningjiang, LIU Xiaoli. Effects of TNF-α signal pathway in pathogenesis of asthma in children [J]. Journal of Jilin University Medicine Edition, 2015, 41(02): 379-382.
[5]	. Enzyme properties and classification of Neanthes japonica (Iznka) fibrinolytic enzyme [J]. J4, 2011, 37(2): 267-270.
[6]	ZHANG Xin-mu, HE Jin-chao, YANG Li-li, ZHANG Ying-yu,DONG Wen， HOU Li-zhong,YAN Wei-qun. Hepatoprotective effects of rhKD/APP on acute hepatic injury  [J]. J4, 2007, 33(1): 51-53.