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Freeze-dried human rabies vaccine and its preparation method

A technology for rabies vaccine and rabies virus, which is applied in the field of freeze-dried human rabies vaccine and its preparation, can solve the problems of low antigen expression content, unsatisfactory vaccine yield and quality, and high incidence of side reactions among vaccine recipients, and achieves operation Simple, save raw materials, improve the effect of space utilization

Active Publication Date: 2015-12-09
江生(深圳)生物技术研发中心有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the existing problems of only using bioreactors to produce rabies vaccines, the expression content of effective viral antigens is low, the incidence of side effects of vaccinators is high, and the yield and quality of vaccines cannot meet the standard requirements. The present invention provides a freeze-dried Human rabies vaccine and preparation method thereof

Method used

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  • Freeze-dried human rabies vaccine and its preparation method
  • Freeze-dried human rabies vaccine and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 prepares freeze-dried human rabies vaccine

[0035] (1) The virus seed adopts aG strain rabies virus, and the culture substrate adopts Vero cells;

[0036] (2) Digest and disperse the resuscitated Vero cells with 0.25% trypsin, add 199 culture medium and mix evenly, then add them to a cell factory (CellFactories, Nunc or CORNING, USA) and culture at 37°C until a uniform monolayer of Vero cells is obtained. cell;

[0037] (3) Use 199 culture fluid containing 10% newborn bovine serum to wash and replace the PBS solution in the bioreactor (BIOFLOCELLIGEN310 type, produced by NBS Company of the United States), the bioreactor parameter setting: temperature 37 ° C, rotating speed 50 rpm, pH7.0 , dissolved oxygen 50%, after each parameter is stable, inoculate the monolayer Vero cells that have been digested and dispersed uniformly, and the number of inoculated cells (working volume) is controlled at 2×10 5 A / ml is appropriate;

[0038] (4) According to the grow...

Embodiment 2

[0046] Embodiment 2 prepares freeze-dried human rabies vaccine

[0047] (1) The virus seed adopts aG strain rabies virus, and the culture substrate adopts Vero cells;

[0048] (2) Digest and disperse the resuscitated Vero cells with 0.25% trypsin, add 199 culture medium and mix evenly, then add them to a cell factory (CellFactories, Nunc or CORNING, USA) and culture at 37°C until a uniform monolayer of Vero cells is obtained. cell;

[0049] (3) Use 199 culture fluid containing 10% newborn bovine serum to wash and replace the PBS solution in the bioreactor (BIOFLOCELLIGEN310 type, produced by NBS Company of the United States), the bioreactor parameter setting: temperature 37 ° C, rotating speed 50 rpm, pH7.0 , dissolved oxygen 50%, after each parameter is stable, inoculate the monolayer Vero cells that have been digested and dispersed uniformly, and the number of inoculated cells (working volume) is controlled at 3×10 5 It is advisable to be within the range of / ml;

[0050]...

Embodiment 3

[0058] Embodiment 3 prepares freeze-dried human rabies vaccine

[0059] (1) The virus seed adopts aG strain rabies virus, and the culture substrate adopts Vero cells;

[0060] (2) Digest and disperse the resuscitated Vero cells with 0.25% trypsin, add 199 culture medium and mix evenly, then add them to a cell factory (CellFactories, Nunc or CORNING, USA) and culture at 37°C until a uniform monolayer of Vero cells is obtained. cell;

[0061] (3) Use 199 culture fluid containing 10% newborn bovine serum to wash and replace the PBS solution in the bioreactor (BIOFLOCELLIGEN310 type, produced by NBS Company of the United States), the bioreactor parameter setting: temperature 37 ° C, rotating speed 50 rpm, pH7.0 , dissolved oxygen 50%, after each parameter is stable, inoculate the single layer of Vero cells that have been digested and dispersed uniformly, and the number of inoculated cells (working volume) is controlled at 4×10 5 It is advisable to be within the range of / ml;

...

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Abstract

The invention relates to a freeze-dried rabies vaccine for humans and a preparation method of the vaccine, relates to the field of vaccine production preparation technologies and aims at solving the problems that effective virus antigen expression content is low, the side effect rate of a vaccinator is high and vaccine yield and quality can not meet standard requirements as only a biological reactor is adopted for producing a rabies vaccine. The freeze-dried rabies vaccine for humans is obtained by inoculating aG strain rabies virus on Vero cells and sequentially carrying out ultrafiltration and concentration, separation and purification as well as freeze drying, wherein the packing volume of the freezed-dried rabies vaccine for human use is 0.5ml / dose, and during freeze drying, the adopted vaccine freeze-drying protecting agent comprises the following ingredients: 60-90g / l of trehalos, 6-14g / l of sodium glutamate, 3-6g / l of urea, 2-3g / l of L-arginine and 10g / l of 199 culture medium, and the vaccine freeze-drying protecting agent does not contain gelatin, human serum albumin or dextran. The freeze-dried rabies vaccine for humans has the advantages that cost is low, operation is easy, pollution is hardly produced, vaccine quality and yield are greatly improved, the content of impurities in a vaccine is reduced, allergy reactions are hardly caused, and vaccine safety is greatly improved.

Description

technical field [0001] The invention belongs to the technical field of vaccine production and preparation, and in particular relates to a freeze-dried human rabies vaccine and a preparation method thereof. Background technique [0002] Rabies (Rabies) is an acute infectious disease caused by rabies virus, which is a natural foci or animal origin zoonotic. The typical clinical manifestation of rabies is hydrophobia, so rabies is also called hydrophobia. Sensitivity to sound, light, wind and other stimuli in the early stage and tightness in the throat. In the excited stage, it may manifest as extreme terror, fear of water, fear of wind, paroxysmal pharyngeal muscle spasm, dyspnea, etc., and finally the spasm stops and various symptoms appear. A paralysis that can rapidly lead to death from respiratory and circulatory failure. Rabies is mainly caused by biting, scratching or mucosal infection of sick animals. Under certain conditions, it can also be transmitted through respir...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/205A61P31/14C12N7/04
Inventor 李锦宇张欢张晶鞠长军王群孙婵池凤臣赵志平于磊张愉莫飞赵爽孙婷婷赵洪洋张秋菊
Owner 江生(深圳)生物技术研发中心有限公司
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