Journal of East China Normal University(Natural Science) ›› 2021, Vol. 2021 ›› Issue (3): 114-127.doi: 10.3969/j.issn.1000-5641.2021.03.012
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Hongwei SHEN, Minghao LI, Nan XU, Jiaqi SHAO, Jing WANG, Lei YU*()
Received:
2019-12-25
Online:
2021-05-25
Published:
2021-05-26
Contact:
Lei YU
E-mail:ylyh188@163.com
CLC Number:
Hongwei SHEN, Minghao LI, Nan XU, Jiaqi SHAO, Jing WANG, Lei YU. Preparation and stability study of lyophilized lentiviral vector[J]. Journal of East China Normal University(Natural Science), 2021, 2021(3): 114-127.
Table 1
Effects of different lyophilized protectants"
分组 | 冻干保护剂处方 |
Ⅰ | HBSS |
Ⅱ | 15% 海藻糖、0.010 mol/L L-组氨酸、0.010 mol/L L-丙氨酸、0.10 g/L CaCl2、0.076 g/L MgSO4 |
Ⅲ | 3.0% 海藻糖、3.0% 甘露醇、2.0% 右旋糖酐、1.5% 肌醇 |
Ⅳ | 0.30% 人血白蛋白、2.0% 海藻糖、1.0% 甘露醇、2.0% 右旋糖酐、2.0% 蔗糖 |
Ⅴ | 1.0% 甘油、20 mg/mL 蔗糖、25 mg/mL 甘露醇、1.0 mg/mL L-精氨酸、 2.0 mg/mL 甘氨酸、2.0% PEG6000、1.5% 明胶 |
Table 3
Orthogonal experimental design table"
实验号 | 海藻糖/g | L-丙氨酸/mg | L-组氨酸/mg | CaCl2/mg | MgSO4/mg |
1 | 3.750 | 8.900 | 15.50 | 25.00 | 0.7600 |
2 | 15.00 | 222.5 | 15.50 | 25.00 | 0.7600 |
3 | 15.00 | 8.900 | 387.5 | 25.00 | 0.7600 |
4 | 9.375 | 115.7 | 201.5 | 13.00 | 9.880 |
5 | 15.00 | 222.5 | 15.50 | 1.000 | 19.00 |
6 | 3.750 | 222.5 | 15.50 | 25.00 | 19.00 |
7 | 15.00 | 8.900 | 387.5 | 1.000 | 19.00 |
8 | 9.375 | 115.7 | 201.5 | 13.00 | 9.880 |
9 | 3.750 | 8.900 | 387.5 | 25.00 | 19.00 |
10 | 3.750 | 8.900 | 15.50 | 1.000 | 19.00 |
11 | 3.750 | 222.5 | 387.5 | 1.000 | 19.00 |
12 | 15.00 | 222.5 | 387.5 | 1.000 | 0.7600 |
13 | 15.00 | 8.900 | 15.50 | 25.00 | 19.00 |
14 | 15.00 | 8.900 | 15.50 | 1.000 | 0.7600 |
15 | 15.00 | 222.5 | 387.5 | 25.00 | 19.00 |
16 | 9.375 | 115.7 | 201.5 | 13.00 | 9.880 |
17 | 3.750 | 222.5 | 15.50 | 1.000 | 0.7600 |
18 | 3.750 | 8.900 | 387.5 | 1.000 | 0.7600 |
19 | 3.750 | 222.5 | 387.5 | 25.00 | 0.7600 |
Table 6
Results of orthogonal experiment"
实验号 | 海藻糖/g | L-丙氨酸/mg | L-组氨酸/mg | CaCl2/mg | MgSO4/mg | 生物滴度/(IU·mL–1) |
1 | 3.750 | 8.900 | 15.50 | 25.00 | 0.7600 | 2.12 × 107 |
2 | 15.00 | 222.5 | 15.50 | 25.00 | 0.7600 | 2.32 × 107 |
3 | 15.00 | 8.900 | 387.5 | 25.00 | 0.7600 | 2.01 × 107 |
4 | 9.375 | 115.7 | 201.5 | 13.00 | 9.880 | 3.17 × 107 |
5 | 15.00 | 222.5 | 15.50 | 1.000 | 19.00 | 2.16 × 107 |
6 | 3.750 | 222.5 | 15.50 | 25.00 | 19.00 | 2.29 × 107 |
7 | 15.00 | 8.900 | 387.5 | 1.000 | 19.00 | 3.47 × 107 |
8 | 9.375 | 115.7 | 201.5 | 13.00 | 9.880 | 3.20 × 107 |
9 | 3.750 | 8.900 | 387.5 | 25.00 | 19.00 | 4.54 × 107 |
10 | 3.750 | 8.900 | 15.50 | 1.000 | 19.00 | 4.91 × 107 |
11 | 3.750 | 222.5 | 387.5 | 1.000 | 19.00 | 4.22 × 107 |
12 | 15.00 | 222.5 | 387.5 | 1.000 | 0.7600 | 3.89 × 107 |
13 | 15.00 | 8.900 | 15.50 | 25.00 | 19.00 | 2.60 × 107 |
14 | 15.00 | 8.900 | 15.50 | 1.000 | 0.7600 | 5.11 × 107 |
15 | 15.00 | 222.5 | 387.5 | 25.00 | 19.00 | 3.96 × 107 |
16 | 9.375 | 115.7 | 201.5 | 13.00 | 9.880 | 4.09 × 107 |
17 | 3.750 | 222.5 | 15.50 | 1.000 | 0.7600 | 2.15 × 107 |
18 | 3.750 | 8.900 | 387.5 | 1.000 | 0.7600 | 2.81 × 107 |
19 | 3.750 | 222.5 | 387.5 | 25.00 | 0.7600 | 1.46 × 107 |
Table 7
Analysis of variance of orthogonal experiment"
来源 | 自由度 | Adj SS | Adj MS | F 值 | P 值 |
模型 | 12 | 2.035 45 × 1015 | 1.696 21 × 1014 | 189.23 | 0 |
线性 | 5 | 8.070 11 × 1014 | 1.614 02 × 1014 | 180.06 | 0 |
海藻糖 | 1 | 6.563 38 × 1012 | 6.563 38 × 1012 | 7.320 0 | 0.042 |
L-丙氨酸 | 1 | 1.645 09 × 1014 | 1.645 09 × 1014 | 183.53 | 0 |
L-组氨酸 | 1 | 4.521 50 × 1013 | 4.521 50 × 1013 | 50.440 | 0.001 |
CaCl2 | 1 | 3.439 20 × 1014 | 3.439 20 × 1014 | 383.67 | 0 |
MgSO4 | 1 | 2.468 04 × 1014 | 2.468 04 × 1014 | 275.33 | 0 |
2 因子交互作用 | 7 | 1.228 44 × 1015 | 1.754 92 × 1014 | 195.78 | 0 |
海藻糖 × L-丙氨酸 | 1 | 7.207 12 × 1013 | 7.207 12 × 1013 | 80.400 | 0 |
海藻糖 × MgSO4 | 1 | 4.591 37 × 1014 | 4.591 37 × 1014 | 512.21 | 0 |
L-丙氨酸 × L-组氨酸 | 1 | 2.660 03 × 1014 | 2.660 03 × 1014 | 296.75 | 0 |
L-丙氨酸 × CaCl2 | 1 | 4.330 05 × 1013 | 4.330 05 × 1013 | 48.310 | 0.001 |
L-组氨酸 × CaCl2 | 1 | 4.194 36 × 1013 | 4.194 36 × 1013 | 46.790 | 0.001 |
L-组氨酸 × MgSO4 | 1 | 2.089 88 × 1014 | 2.089 88 × 1014 | 233.15 | 0 |
CaCl2 × MgSO4 | 1 | 1.369 98 × 1014 | 1.369 98 × 1014 | 152.83 | 0 |
误差 | 5 | 4.481 91 × 1012 | 8.963 83 × 1011 | ||
失拟 | 4 | 4.438 46 × 1012 | 1.109 61 × 1012 | 25.530 | 0.147 |
纯误差 | 1 | 4.345 75 × 1010 | 4.345 75 × 1010 | ||
合计 | 17 | 2.039 93 × 1015 |
1 |
NALDINI L, TRONA D, VERMA I M. Lentiviral vectors, two decades later. Science, 2016, 353 (6304): 1101- 1102.
doi: 10.1126/science.aah6192 |
2 |
MILONE M C, O DOHERTY U. Clinical use of lentiviral vectors. Leukemia, 2018, 32 (7): 1529- 1541.
doi: 10.1038/s41375-018-0106-0 |
3 |
DUNBAR C E, HIGH K A, JOUNG J K, et al. Gene therapy comes of age. Science, 2018, 359 (6372): n4672.
doi: 10.1126/science.aan4672 |
4 |
CARMO M, ALVES A, RODRIGUES A F, et al. Stabilization of gammaretroviral and lentiviral vectors: from production to gene transfer. The Journal of Gene Medicine, 2009, 11 (8): 670- 678.
doi: 10.1002/jgm.1353 |
5 | YANNARELL D A, GOLDBERG K M, HJORTH R N. Stabilizing cold-adapted influenza virus vaccine under various storage conditions. Journal of Virological Methods, 2002, 102 (1): 15- 25. |
6 |
KUMRU O S, WANG Y, GOMBOTZ C W R, et al. Physical Characterization and Stabilization of a Lentiviral Vector Against Adsorption and Freeze-Thaw. Journal of Pharmaceutical Sciences, 2018, 107 (11): 2764- 2774.
doi: 10.1016/j.xphs.2018.07.010 |
7 |
李仲艺. 真空冷冻干燥技术在生物制药方面的应用分析. 中国新技术新产品, 2018, (1): 76- 77.
doi: 10.3969/j.issn.1673-9957.2018.01.042 |
8 | 耿锟锟, 熊非, 朱家壁, 等. 用于药物制剂的冷冻干燥技术及相关影响因素. 药学进展, 2011, 35 (3): 104- 109. |
9 |
田烨, 吴明媛. 生物制品冻干保护方法研究进展. 中国医药生物技术, 2018, 13 (1): 73- 76.
doi: 10.3969/j.issn.1673-713X.2018.01.015 |
10 |
HANSEN L J J, DAOUSSI R, VERVAET C, et al. Freeze-drying of live virus vaccines: A review. Vaccine, 2015, 33 (42): 5507- 5519.
doi: 10.1016/j.vaccine.2015.08.085 |
11 |
MORGAN C A, HERMAN N, WHITE P A, et al. Preservation of micro-organisms by drying: A review. Journal of Microbiological Methods, 2006, 66 (2): 183- 193.
doi: 10.1016/j.mimet.2006.02.017 |
12 |
DE LAS MERCEDES SEGURA M, KAMEN A, GARNIER A. Downstream processing of oncoretroviral and lentiviral gene therapy vectors. Biotechnology Advances, 2006, 24 (3): 321- 337.
doi: 10.1016/j.biotechadv.2005.12.001 |
13 |
KUTNER R H, ZHANG X, REISER J. Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors. Nature Protocols, 2009, 4 (4): 495- 505.
doi: 10.1038/nprot.2009.22 |
14 |
TISCORNIA G, SINGER O, VERMA I M. Production and purification of lentiviral vectors. Nature Protocols, 2006, 1 (1): 241- 245.
doi: 10.1038/nprot.2006.37 |
15 | 上海比昂生物医药科技有限公司. 一种慢病毒冷冻干燥保护剂及慢病毒冻干粉: CN201810995003.1 [P]. 2018-10-09. |
16 |
张一折, 姜春来, 王忠诚, 等. 重组MVA病毒载体疫苗的冻干保护剂研究. 中国生物制品学杂志, 2006, (2): 174- 176.
doi: 10.3969/j.issn.1004-5503.2006.02.021 |
17 |
张一折, 滕洪刚, 吕帅然, 等. 重组HIV-1腺病毒载体活疫苗的冻干保护剂研究. 中国生物制品学杂志, 2007, (2): 104- 106.
doi: 10.3969/j.issn.1004-5503.2007.02.010 |
18 |
ADEBAYO A A, SIM-BRANDENBURG J W, EMMEL H, et al. Stability of 17D yellow fever virus vaccine using different stabilizers. Biologicals, 1998, 26 (4): 309- 316.
doi: 10.1006/biol.1998.0157 |
19 |
PRABHU M, BHANUPRAKASH V, VENKATESAN G, et al. Evaluation of stability of live attenuated camelpox vaccine stabilized with different stabilizers and reconstituted with various diluents. Biologicals, 2014, 42 (3): 169- 175.
doi: 10.1016/j.biologicals.2014.02.001 |
20 |
SARKAR J, SREENIVASA B P, SINGH R P, et al. Comparative efficacy of various chemical stabilizers on the thermostability of a live-attenuated peste des petits ruminants (PPR) vaccine. Vaccine, 2003, 21 (32): 4728- 4735.
doi: 10.1016/S0264-410X(03)00512-7 |
21 |
SIVA SANKAR M S, BHANUPRAKASH V, VENKATESAN G, et al. Comparative efficacy of chemical stabilizers on the thermostabilization of a novel live attenuated buffalopox vaccine. Biologicals, 2017, 49, 39- 45.
doi: 10.1016/j.biologicals.2017.07.002 |
22 | 马海燕, 方彧聃, 张敬之. 应用荧光实时定量PCR方法检测重组慢病毒滴度及其感染效率. 生命科学研究, 2009, 13 (5): 394- 398. |
23 |
玛尔江·木坎, 卢琳. 卡尔费休氏试剂与水分测定. 畜禽业, 2016, (5): 42- 43.
doi: 10.3969/j.issn.1008-0414.2016.05.032 |
24 |
傅晖, 顾菁. 腺病毒冻干条件的改进研究. 上海医药, 2017, 38 (13): 67- 69.
doi: 10.3969/j.issn.1006-1533.2017.13.019 |
25 |
HAN Y, JIN B S, Lee S B, et al. Effects of sugar additives on protein stability of recombinant human serum albumin during lyophilization and storage. Arch Pharm Res, 2007, 30 (9): 1124- 1131.
doi: 10.1007/BF02980247 |
26 | 张光磊, 张新创, 翟雷. 活菌制剂冻干保护剂的研究进展. 微生物学免疫学进展, 2015, 43 (4): 80- 85. |
27 | 薛菲, 王凤山. 蛋白质的冻干保护剂及其保护机制研究进展. 中国药学杂志, 2018, 53 (10): 765- 770. |
28 |
OLSSON C, JANSSON H, SWENSON J. The Role of Trehalose for the Stabilization of Proteins. The Journal of Physical Chemistry B, 2016, 120 (20): 4723- 4731.
doi: 10.1021/acs.jpcb.6b02517 |
29 |
ZHANG M, OLDENHOF H, SYDYKOV B, et al. Freeze-drying of mammalian cells using trehalose: preservation of DNA integrity. Scientific Reports, 2017, 7 (1): 6198.
doi: 10.1038/s41598-017-06542-z |
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