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Preparation of recombinant adenovirus

A recombinant adenovirus and preparation technology, which is applied in the field of preparations, can solve problems such as the inability to use adenovirus preparations and the inability to maintain the activity of recombinant adenoviruses, and achieve good stability.

Active Publication Date: 2011-04-27
北京康弘生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these improved preparations cannot be used to preserve adenovirus preparations with a concentration higher than 2E12vp / ml, and cannot maintain the activity of recombinant adenovirus in the preparation for a long period of time (more than 2 years) at 2-8°C

Method used

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  • Preparation of recombinant adenovirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Effect of pH value on the stability of recombinant adenovirus in preparation

[0046] In order to study the effect of pH value on the stability of recombinant adenovirus in the preparation, at 25°C, we studied the range of pH 6.8-8.6, and the pH values ​​were 6.8, 7.2, 7.4, 7.6, 7.8, 8.2, and 8.6. , the effect of pH on the stability of recombinant adenoviruses in preparations.

[0047] By detecting the virus titer (TCID50 / mL) and specific activity of the recombinant adenovirus in the preparation, the effect of pH value on the stability of the recombinant adenovirus in the preparation was studied, and the results are shown in Table 1.

[0048] The influence of table 1pH value on the stability of recombinant adenovirus in the preparation

[0049]

[0050] Note: The composition of the preparation is 10mM Tris+10w / v% propylene glycol, and the pH is adjusted with HCl.

[0051] It can be seen from Table 1 that when the pH is lower than 7.4, the recombinant ad...

Embodiment 2

[0052] Example 2 Effect of protective agent on the stability of recombinant adenovirus in preparation

[0053] The effects of protective agents (propylene glycol, glycerol, polyethylene glycol 400, dimethyl sulfoxide, sorbitol, mannitol, sucrose, trehalose) on the stability of recombinant adenovirus in the preparation were studied at 25°C . Wherein, the control preparation is ARCA preparation.

[0054] By detecting the virus titer (TCID50 / mL) and specific activity of the recombinant adenovirus in the preparation, the influence of the protective agent on the stability of the recombinant adenovirus preparation in the preparation was studied. The composition and research results of the preparation are shown in Table 2.

[0055] The impact of table 2 protective agent on the stability of recombinant adenovirus in the preparation

[0056]

[0057]

[0058] Note: The Tris concentration in the Tris-HCl buffer solution is 10mM, and the pH is adjusted to 7.8 with HCl during pr...

Embodiment 3

[0060] Example 3 Effect of Amino Acids on the Stability of Recombinant Adenovirus in Preparation

[0061] Under the condition of 25°C, the present invention studies the influence of amino acids (glycine, histidine, lysine, arginine, methionine, proline) on the stability of the recombinant adenovirus in the preparation.

[0062] By detecting the virus titer (TCID50 / mL) and specific activity of the recombinant adenovirus in the preparation, the influence of amino acids on the stability of the recombinant adenovirus preparation in the preparation was studied. The composition and research results of the preparation are shown in Table 3.

[0063] The impact of table 3 amino acids on the stability of recombinant adenovirus in the preparation

[0064]

[0065] Note: Tris concentration in Tris-HCl buffer is 10mM, adjust pH to 7.8 with HCl during preparation; propylene glycol concentration is 10w / v%.

[0066] It can be seen from Table 3 that compared with the preparation without ...

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Abstract

The invention relates to a preparation of recombinant adenovirus, containing recombinant adenovirus, Tris buffer solution, propylene glycol and arginine; in addition, the preparation also can contain an antioxidant, such as one or combination of sodium sulfite, sodium thiosulfate, glutathione, thiosalicylic acid, cystine hydrochloride, dithioglycollil acid, thiourea, sulfosorbierite or aurothioglucose; the preparation also can contain antioxidant synergist which is one or combination of sodium citrate, sodium succinate or sodium hypophosphite. By utilizing the preparation, the recombinant adenovirus can be stored for long time at the temperature of 2-8 DEG C.

Description

technical field [0001] The invention relates to a preparation, in particular to a preparation of recombinant adenovirus. Background technique [0002] Recombinant adenovirus is a commonly used replication carrier in the field of gene therapy. It is required for clinical use to maintain a complete icosahedral virus capsid structure during storage, transportation and use, and to maintain a high number of live viruses and infection and replication capabilities. However, adenoviruses are extremely unstable and easily degraded under conditions such as high temperature, low pH, and oxidation. For example, US7202078 discloses the following technical content, when the concentration of recombinant adenovirus is higher than 1E12vp / ml, virus particles will usually aggregate into filaments, triggering capsid protein cleavage, resulting in reduced infection activity. [0003] In actual clinical application, the traditional method mainly chooses to store and transport recombinant adenovi...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K47/18A61K47/10
Inventor 杨艳艳郭勇
Owner 北京康弘生物医药有限公司
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