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Grippe primary generation susliks kidney cell multivalent vaccine and its preparation method

A technology of influenza and hamster kidney cells, which is applied in pharmaceutical formulas, medical preparations containing active ingredients, inactivation/attenuation, etc., to achieve the effects of fast growth, slow resolution and adaptation, and sufficient kidney sources

Inactive Publication Date: 2013-04-17
深圳市孚沃德生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

No one at home and abroad has reported the precedent of using primary hamster kidney cells to prepare influenza vaccine

Method used

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  • Grippe primary generation susliks kidney cell multivalent vaccine and its preparation method
  • Grippe primary generation susliks kidney cell multivalent vaccine and its preparation method
  • Grippe primary generation susliks kidney cell multivalent vaccine and its preparation method

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preparation example Construction

[0055] 1. Preparation of Hamster Kidney Cells

[0056] Choose healthy golden hamsters aged 10-14 days, kill them with drinking water, wash them 1-2 times, and disinfect them with 1‰ bromogeramine for 1-3 times, each time for 3-8 minutes. In a sterile environment, use sterile scissors to dissect the hamster and take out the kidney, cut it into pieces, add a digestive solution composed of 0.1% to 0.5% trypsin and 0.01% to 0.05% EDTA, and place it in a cold place at 2 to 8°C. Digest for 15-20 hours, discard the digestion solution, add growth solution to disperse the cells, and prepare 1.0×10 7 ~1.0×10 8 cells / ml of cell suspension. Take the primary cell suspension and inoculate it in a 3L or 10L spinner bottle or in a cell bioreactor according to the ratio of cell suspension: growth liquid of 1:20 to 1:100, and then add growth liquid to make the cell The initial concentration is 1.0×10 5 ~5.0×10 6 pieces / ml. Spinner bottle at 37°C with CO 2 The cell culture is carried out ...

Embodiment 1

[0061] Example 1: Preparation of virus seeds adapted to primary hamster kidney cells of influenza virus

[0062] Primal Poison Seed Armor 1 (H 1 N 1 ) type is IVR-116, which is the 8th generation chicken embryo allantoic fluid freeze-dried and preserved virus species; 3 (H 3 Type N2) is NYMC X-15F, which is the 8th generation chicken embryo allantoic fluid freeze-dried preservation virus species; B type is B / Jiangsu / 10 / 2003, which is the 6th generation chicken embryo allantoic fluid freeze-dried preservation virus species.

[0063] The above-mentioned three virus species were unsealed in a special aseptic room, and adaptive passages were carried out on SPF chicken embryos for 2 times. The finally harvested viral allantoic fluid was subjected to sterility test and hemagglutination titer (HA titer) measurement respectively. The sterility test is qualified, the hemagglutination titer of A1 and A3 are both 1:640, and the hemagglutination titer of type B is 1:320, thus establi...

Embodiment 2

[0068] Example 2: Preparation of virus seeds adapted to primary hamster kidney cells of influenza virus

[0069] Primal Poison Seed Armor 1 (H 1 N 1 ) type is IVR-116, which is the 8th generation chicken embryo allantoic fluid freeze-dried preservation virus species: A 3 (H 3 N 2) type is NYMC X-15F, which is lyophilized and preserved in the 8th generation of chicken embryo allantoic fluid; type B is B / Jiangsu / 10 / 2003 (recommended by WHO in 2004), which is lyophilized and preserved in the 6th generation of chicken embryo allantoic fluid Poison species.

[0070] The main generation seeds are the three types of influenza main generation seeds prepared in Example 1.

[0071] The anatomy of the hamster kidney, the preparation of the cell suspension, and the inoculation of the glass bottle for culture are the same as in Example 1.

[0072] When the cells in the culture flask occupy 60% to 80% of the culture surface, replace with maintenance solution I after rinsing, and use ...

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Abstract

A purified multi-valent vaccine of influenza made of the kidney cells of primary vole is disclosed. Its preparing process includes such steps as preparing the viral culture base from the kidney cells of primary vole, culturing the current viruses of human or fowl's influenza, adaptive reproduction and passage to obtain influenza virus strains, culturing the said kidney cells infected by current virus to obtain the primary liquid of influenza virus, centrifugal separation, ultrafiltration, deactivating, ultra-speed centrifugal treating, and purifying by column chromatography.

Description

technical field [0001] The invention relates to a primary influenza hamster kidney cell vaccine and a preparation method thereof, belonging to the field of biopharmaceuticals. Background technique [0002] Influenza is called flu for short, is a kind of serious respiratory infectious disease. In the influenza pandemic of 1918-1919, more people died due to infection with the influenza virus than in World War II, which lasted 4 years. Influenza is often characterized by repeated epidemics, sudden onset, and rapid spread, ranging from small local epidemics to large ones all over the world. Infants and the elderly have a high case fatality rate of tens of thousands every year. At present, influenza is still an important issue affecting public health in the world, and the recent highly pathogenic avian influenza has a great danger of invading humans, so vaccination is still one of the important means to prevent influenza and avian influenza infection. [0003] As the causative...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/145C12N7/08A61P11/00A61P31/16
Inventor 李云英王玉清吴歧
Owner 深圳市孚沃德生物技术有限公司
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