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Preparation method of recombinant adenovirus preparation
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A recombinant adenovirus and preparation technology, which is applied in the field of preparation of recombinant adenovirus preparations, to achieve the effects of good safety and stability, long validity period and good stability
Inactive Publication Date: 2013-02-06
天津泽世德生物医药有限公司
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[0003] The technical problem to be solved by the present invention is to solve the stability of existing recombinant ade
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[0019] Mix the components according to the formula, add pure water to 100ml, and then add the recombinant adenovirus according to the amount of adenovirus contained in it 1.0×10 12 The number of particles / ml is added to the above protective agent at a concentration of 1:0.8, mixed evenly, sterile filtered, filled, and vacuum freeze-dried, and the preparation is placed in a freeze-drying box, and the relative pressure in the freeze-dryer is reduced to 5Pa, preset heating program, maintain 30 minutes at 15°C each time, continue depressurization and increase temperature to 0-35°C and maintain for 3 hours to prepare freeze-dried recombinant adenovirus preparations.
[0023] Mix the components according to the formula, add pure water to 100ml, and then add the recombinant adenovirus according to the amount of adenovirus contained in it 1.0×10 12 The number of particles / ml is added to the above protective agent at a concentration of 1:0.8, mixed evenly, sterile filtered, filled, and vacuum freeze-dried, and the preparation is placed in a freeze-drying box, and the relative pressure in the freeze-dryer is reduced to 5Pa, preset heating program, maintain 30 minutes at 15°C each time, continue depressurization and increase temperature to 0-35°C and maintain for 3 hours to prepare freeze-dried recombinant adenovirus preparations.
Embodiment 3
[0025] Protective agent formula: hydroxyethyl starch 19.8g, arginine 1.7g, glycine 0.2g, human serum albumin 10ml, trehalose 5.9g
[0026] preparation:
[0027] Mix the components according to the formula, add pure water to 100ml, and then add the recombinant adenovirus according to the amount of adenovirus contained in it 1.0×10 12 The number of particles / ml is added to the above protective agent at a concentration of 1:1.1, mixed evenly for filling, and the vacuum freeze-drying method is used to put the preparation into a freeze-drying box. After the relative pressure in the freeze-dryer drops to 5Pa, the preset The temperature raising program is to raise the temperature by 15°C each time for 30 minutes, and continue to reduce the pressure to raise the temperature to 0-35°C and maintain it for 3 hours to prepare a freeze-dried recombinant adenovirus preparation.
[0028] The freeze-dried recombinant adenovirus preparation made by the method of the present invention, the inf...
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Abstract
The invention discloses a preparation method of a recombinant adenovirus preparation. The invention relates to a field of lyophilized preparation and a preparation method thereof. The invention aims at providing a recombinant adenovirus preparation. A technical scheme provided by the invention is that: the recombinant adenovirus preparation comprises a recombinant adenovirus stock solution and a lyophilization additive. The lyophilization additive is substantially composed of human serum albumin and hydroxyethyl starch. The method for preparing the recombinant adenovirus preparation comprises the steps that: (1) the recombinant adenovirus stock solution and the lyophilization additive are well mixed; the mixture is subjected to sterile filtration; and the mixture is sub-packaged into vials; (2) the vials are placed into a lyophilizing device, and are respectively equilibrated for 1h under temperatures of 5 DEG C, 0 DEG C, and -6 DEG C; (3) rapid freezing is carried out, wherein a product temperature is reduced to -30 DEG C to -40 DEG C, and the temperature is maintained for 3h; (4) vacuum drying is carried out; (5) heating drying is carried out, wherein a temperature is maintained for 3-5h when the product temperature reaches 0 DEG C to 35 DEG C; and (6) the products are plugged and sealed.
Description
technical field [0001] The invention relates to a preparation method of a recombinant adenovirus preparation, in particular to a preparation method of a recombinant adenovirus preparation. Background technique [0002] Adenovirus has become a research hotspot of viral vectors in recent years because of its low pathogenicity to humans, no induction of carcinogenesis, wide range of host cells, high reproductive titer, and large loading capacity. However, even liquid recombinant adenovirus preparations containing stabilizers are relatively sensitive to temperature, which brings great difficulties to transportation and storage, which largely limits their clinical application. So far, there is no freeze-dried recombinant adenovirus preparation that can be stored at 4-8°C for a long time on the market. Contents of the invention [0003] The technical problem to be solved by the present invention is to solve the stability of the existing recombinant adenovirus preparation, and p...
Claims
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