Freeze-thaw method for single virus harvest

A virus and a single technology, applied in the field of freezing and thawing of a single virus harvest solution, can solve problems such as the inability to wait for the completion of the test, and achieve the effect of reducing the loss of virus titer, reducing the risk of discarding, and prolonging the storage time

Active Publication Date: 2022-07-19
CHENGDU INST OF BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Utilizing the freezing storage conditions in this document (storage at 2-8°C), after the titer test results of a single virus harvest liquid come out, it is necessary to put the single virus harvest liquid into production as soon as possible. Assay completed, leading to risk release of intermediate product

Method used

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  • Freeze-thaw method for single virus harvest
  • Freeze-thaw method for single virus harvest
  • Freeze-thaw method for single virus harvest

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: Freeze-thaw method of single virus harvest liquid of the present invention

[0031] (1) Cryopreservation

[0032] Freeze 14L of Japanese encephalitis live attenuated vaccine single virus harvest liquid in a freezing bag to -70 ℃ for 120 days;

[0033] (2) Melting

[0034] After 120 days of cryopreservation, the freezing bag containing the single virus harvest solution of live attenuated Japanese encephalitis vaccine was thawed in a constant temperature water bath at 27±2°C, and thawed completely after 30 minutes.

[0035] The beneficial effects of the present invention are demonstrated below through experimental examples.

experiment example 1

[0036] Experimental Example 1: Screening Experiment of Cryopreservation Medium

[0037] 1. Experimental method

[0038] According to the freezing and thawing method of Example 1, the only difference is that: in the freezing step, glass bottles, plastic steel bottles, and freezing bags are respectively used for freezing, and the influence of freezing medium is studied.

[0039] 2. Experimental results

[0040] The experiment found that if a single virus harvest solution is frozen in a glass bottle, the glass bottle is easily damaged during the freezing and thawing process, and it is not suitable for the freezing and thawing process;

[0041] The advantages of freezing a single virus harvest solution in plastic cylinders are that it can be reused and the cost is low.

[0042] The advantages of freezing a single virus harvest solution in a freezing bag are that it is easy to freeze and thaw, and it is easy to store. The disadvantage is that it is one-time use and the cost is h...

experiment example 2

[0043] Experimental Example 2: Screening Experiment of Cryopreservation Conditions

[0044] 1. Experimental method

[0045] According to the freezing and thawing method of Example 1, the difference is only that: in the freezing storage step, the freezing temperature and freezing time shown in Table 2 are respectively used for freezing storage, and the effects of different freezing storage conditions on the titer of single virus harvest are compared. .

[0046] 2. Experimental results

[0047] Table 2 The effect of different freezing conditions on the titer of a single virus harvest

[0048]

[0049]

[0050] It can be seen that the titer of a single virus harvest solution drops significantly after thawing after 120 days of freezing at -20 °C, and this method is not applicable; if it is frozen at -40 °C for 120 days, the drop of a single virus harvest solution after thawing is not applicable. The degree of decrease is obvious, this method is not applicable.

[0051] H...

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Abstract

The invention discloses a method for freezing and thawing a single virus harvest liquid, which belongs to the field of vaccine preparation technology. The freezing and thawing method includes the following steps: (1) cryopreservation: freezing a single virus harvest solution; (2) thawing: thawing the single virus harvest solution after freezing in step (1). Freezing and thawing the single virus harvest solution of the live attenuated Japanese encephalitis vaccine under the freeze-thaw method of the present invention can enable the single virus harvest solution to be stored for a long time under the condition of ≤-60°C, and then put into the next step after all the verification items are completed. Use, for the preparation of Japanese encephalitis live attenuated vaccine. By combining the single virus harvest solution of the Japanese encephalitis live attenuated vaccine of the present invention with the freezing-thawing method of the present invention, the storage time can be significantly prolonged, and the loss of virus titer can also be significantly reduced, and there is no need to carry out the risk release of intermediate products. The arrangement is more flexible and controllable, and the risk of a single virus harvest liquid being discarded due to changes in production plans is greatly reduced.

Description

technical field [0001] The invention belongs to the field of vaccine preparation technology, and particularly relates to a method for freezing and thawing a single virus harvest liquid. Background technique [0002] Live attenuated Japanese encephalitis vaccine (live Japanese encephalitis vaccine) can stimulate the body to produce immunity against Japanese encephalitis virus and is used to prevent Japanese encephalitis. 2. The attenuated JE strain was inoculated into primary SPF hamster kidney cells, and the virus was cultured, harvested, added with a suitable protective agent, packaged, and freeze-dried. [0003] In the production of live JE vaccine, viral antigens (ie, single virus harvest solution) are periodically produced in a continuous, batch-by-batch manner, and are used in batches for the preparation of semi-finished vaccine products. According to the provisions of Part III of the Chinese Pharmacopoeia 2015, the single virus harvest liquid needs to be sampled and c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00C12R1/93
CPCC12N7/00C12N2770/24151
Inventor 刘少祥郭红金鹏李军王玖强周超李俊明
Owner CHENGDU INST OF BIOLOGICAL PROD
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