Freeze thawing method of single virus harvest liquid

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: 2022-01-18
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 thawing method of single virus harvest liquid
  • Freeze thawing method of single virus harvest liquid
  • Freeze thawing method of single virus harvest liquid

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) Frozen storage

[0032] Freeze the 14L live attenuated Japanese encephalitis vaccine single virus harvest solution in a freezer bag to -70°C for 120 days;

[0033] (2) melting

[0034] After 120 days of frozen storage, the frozen storage bag containing the single virus harvest liquid of the live attenuated Japanese encephalitis vaccine was placed in a constant temperature water bath at 27±2°C to thaw, and it was completely thawed after 30 minutes.

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

experiment example 1

[0036] Experimental example 1: Screening experiment of freezing medium

[0037] 1. Experimental method

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

[0039] 2. Experimental results

[0040] Experiments have found that using glass bottles to freeze a single virus harvest solution, the glass bottles are easily damaged during the freeze-thaw process, and are not suitable for the freeze-thaw process;

[0041] Freezing a single virus harvest solution in a plastic steel bottle has the advantage of being reusable and low in cost; the disadvantage is that it takes up a lot of storage space, and when thawing in a water bath, the contact with the water bath is insufficient and the thawing time is long;

[0042] The advantage of using a ...

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 only difference is that in the freezing storage step, the freezing storage temperature and freezing storage time shown in Table 2 are respectively used for freezing storage, and the influence of different freezing storage conditions on the titer of a single virus harvested liquid is compared .

[0046] 2. Experimental results

[0047] Table 2 The impact 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 harvested liquid drops significantly after thawing at -20°C for 120 days, and this method is not applicable; when frozen at -40°C for 120 days, the titer of a single virus harvested liquid after thawing drops significantly. The speed drops obviously, this method is not applicable.

[0051] ...

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Abstract

The invention discloses a freeze thawing method of a single virus harvest liquid, and belongs to the field of vaccine preparation processes. The freeze thawing method comprises the following steps: (1) cryopreservation: cryopreserving a single virus harvesting liquid; and (2) melting: melting the single virus harvesting liquid after cryopreservation in the step (1). The single virus harvest liquid of the Japanese encephalitis attenuated live vaccine is frozen and thawed under the freezing and thawing method disclosed by the invention, so that the single virus harvest liquid can be stored for a long time under the condition that the temperature is less than or equal to-60 DEG C, and after all verification items are finished, the single virus harvest liquid is put into the next step to be used for preparing the Japanese encephalitis attenuated live vaccine. When the single virus harvest liquid of the Japanese encephalitis live attenuated vaccine is matched with the freezing and thawing method, the storage time is remarkably prolonged, meanwhile, the virus titer loss can be remarkably reduced, risk release of intermediate products is not needed, production arrangement is more flexible and controllable, the risk that a single virus harvest liquid is abandoned due to the change of a production plan is also greatly reduced.

Description

technical field [0001] The invention belongs to the field of vaccine preparation technology, and in particular relates to a method for freezing and thawing a single virus harvest liquid. Background technique [0002] Japanese encephalitis live attenuated vaccine (JE live vaccine) can stimulate the body to produce immunity against Japanese encephalitis virus, and is used to prevent epidemic Japanese encephalitis. It is the Japanese encephalitis virus SA14-14- 2 The attenuated JE strain is inoculated with primary SPF hamster kidney cells, cultured, harvested, added with a suitable protective agent, subpackaged, and freeze-dried. [0003] In the production of live JE vaccines, virus antigens (i.e. single virus harvest liquid) are periodically produced continuously and in batches, and are used in batches for the preparation of semi-finished vaccines. According to the provisions of the third part of the Chinese Pharmacopoeia 2015 edition, the single virus harvest liquid needs to...

Claims

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Application Information

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Patent Type & Authority Applications(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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