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Lyophilized preparation of recombinant adenovirus and preparation method thereof

A technology of recombinant adenovirus and freeze-dried preparation is applied in the field of preparation of freeze-dried dosage form products, can solve the problems such as the lack of freeze-dried adenovirus preparation, and can achieve good application prospect, low cost, good safety and stability. Effect

Inactive Publication Date: 2006-12-27
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the application of adenovirus freeze-dried preparations to humans in the field of gene therapy.
So far, there is no freeze-dried adenovirus preparation that can be stored at 4°C-8°C for a long time on the market

Method used

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  • Lyophilized preparation of recombinant adenovirus and preparation method thereof
  • Lyophilized preparation of recombinant adenovirus and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The preparation method of the freeze-dried preparation type recombinant adenovirus is: in 100 milliliters of phosphate physiological sodium chloride solution (PBS) with a pH value of 7.0 to 8.0, add 0.5 grams of human serum albumin, 3 grams of trehalose, and 2 grams of mannitol , 2 grams of dextran, 2 grams of sucrose. After fully dissolving, filter and sterilize with a filter membrane with a pore size of 0.22 μm, and then filter the recombinant adenovirus 10 11 TCID 50 The concentrated solution is added to the above-mentioned solution containing the freeze-drying agent for filling, and after each vial is divided into 0.5 milliliters, vacuum freeze-drying starts. The virus liquid was pre-frozen at -35 to -40°C for 3 to 4 hours, and then dried at a shelf temperature of -30 to -32°C and a vacuum of 10 Pa for 8 to 9 hours. After the product is fully formed, the temperature of the shelf is gradually increased from -10°C, 0°C to 32°C, and then dried under a vacuum of 10Pa ...

Embodiment 2

[0026] The preparation method of the freeze-dried preparation type recombinant adenovirus is: in 100 milliliters of phosphate physiological sodium chloride solution (PBS) with a pH value of 7.0 to 8.0, add 0.2 grams of human serum albumin, 2 grams of trehalose, and 1 gram of mannitol. , 1 gram of dextran, 1 gram of sucrose. After fully dissolving, filter and sterilize with a filter membrane with a pore size of 0.22 μm, and then filter the recombinant adenovirus 2×10 12 TCID 50 The concentrated solution is added to the above-mentioned solution containing the freeze-drying agent for filling, and after each vial is divided into 0.5 milliliters, vacuum freeze-drying starts. The virus liquid was pre-frozen at -35 to -40°C for 3 to 4 hours, and then dried at a shelf temperature of -30 to -32°C and a vacuum of 10 Pa for 8 to 9 hours. After the product is fully formed, the temperature of the shelf is gradually increased from -10°C, 0°C to 32°C, and then dried under a vacuum of 10Pa ...

Embodiment 3

[0029] The preparation method of the freeze-dried preparation type recombinant adenovirus is: in 100 milliliters of phosphate physiological sodium chloride solution (PBS) with a pH value of 7.0 to 8.0, add 0.2 grams of human serum albumin, 4 grams of trehalose, and 3 grams of mannitol. , 2 grams of dextran, 3 grams of sucrose. After fully dissolving, filter and sterilize with a filter membrane with a pore size of 0.22 μm, and then filter the recombinant adenovirus 2×10 13 TCID 50 The concentrated solution is added to the above-mentioned solution containing the freeze-drying agent for filling, and after each vial is divided into 0.5 milliliters, vacuum freeze-drying starts. The virus liquid was pre-frozen at -35 to -40°C for 3 to 4 hours, and then dried at a shelf temperature of -30 to -32°C and a vacuum of 10 Pa for 8 to 9 hours. After the product is fully formed, the temperature of the shelf is gradually increased from -10°C, 0°C to 32°C, and then dried under a vacuum of 10...

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Abstract

Disclosed are a cryodesiccation preparation of recombinant adenoviruses and preparation method thereof, pertaining to biological products technique field, comprising recombinant adenoviruses 2*1012TCID50,human blood albumin 0.2-1g, trehalose 2-4g, mannitol 1-3g, dextran 1-2g, saccharose 1-3g, microcosmic salt physiological natrii chloridum solution, with a pH value of 7.0-8.0, and with a constant volume of 100ml. The recombinant adenoviruses are protected better at a cryodesiccation condition. The preparation is provided with security, economy, ease for production and use, and no shortcoming of strict cold-chain and short period of validity during low temperature freezing storage and transportation as liquid vaccines have.

Description

technical field [0001] The invention belongs to the technical field of biological products, and particularly relates to the preparation of freeze-dried dosage form products using recombinant adenoviruses as expression vectors. Background technique [0002] In the fields of gene therapy and vaccine research, adenoviruses are widely used as vectors for gene transfer. Recombinant adenovirus refers to the insertion of foreign genes into live adenovirus vectors, and the expression of foreign genes in host cells through viruses, thereby stimulating the body to produce a broader and specific immune response. [0003] Recombinant adenoviral vectors are easy to produce, low in cost, less pathogenic to humans, do not induce carcinogenesis, have high reproductive titers, and have large loading capacity, which has become a research hotspot of viral vectors in recent years. However, adenovirus is sensitive to repeated freezing and thawing and is easily inactivated. Due to poor thermal ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/00A61K47/42A61K9/19A61K48/00
Inventor 孔维张一折姜春来滕宏刚吕帅然
Owner JILIN UNIV
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