64results about How to "High viral titer" patented technology

The present invention provides a method for producing a virus vector, which comprises a step wherein cells that are capable of producing a virus vector are cultured in a culture medium that contains, as active components, a retinoic acid and a histone deacetylase inhibiting substance; and a culture medium for the production of a virus vector, which is characterized by containing, as active components, a retinoic acid and a histone deacetylase inhibiting substance.

The invention discloses a technology for the production of porcine epidemic diarrhea virus by means of the microcarrier culture of VREO cells using a bioreactor, and comprises the technology for the production of different porcine epidemic diarrhea virus strains. The technology comprises the following technical steps: (1) selection of VERO cells as cell line for vaccine; (2) passage and culture of cells for vaccine; (3) propagation of seed culture of the porcine epidemic diarrhea virus; (4) microcarrier suspension culture of the VERO cells in the bioreactor; (5) propagation of porcine epidemic diarrhea virus antigen; and (6) treatment of acquired virus antigen liquid. The production method can remarkably lower production cost and enhance output-input ratio by 5 to 10 times, and has the advantages of short production period, small occupied space, great easiness for enlarging production scale rapidly, little environmental pollution, easy processing, high automation degree, a small number of staff, easy implementation of even and stable quality, obviously lowered production cost and enhanced yield and quality of vaccine.

The invention relates to a preparation method of a swine pseudorabies vaccine, which comprises the following steps of: culturing a virus by using a swine testicle cell, and when one layer of cells grows, inoculating a swine pseudorabies virus; then adding into a cell maintenance medium, statically or rotatably culturing, when the cell suffers from more than 80 percent of pathological changes, harvesting a cell culture, repeatedly freeze-thawing to obtain a cell venom containing supernate, and mixing the cell venom qualified in toxic valence detection with formaldehyde for inactivating; and mixing with an emulsifying agent for emulsifying to obtain the swine pseudorabies vaccine. Compared with the prior art, a strain used for preparing the vaccine has the advantages of stronger toxicity and high virus valence; the swine pseudorabies vaccine has good immunogenicity and long immunization period; and the preparation method has the advantages of reasonable process and lower cost, thereby greatly lowering the cost load of the fish breeding and poultry raising industry.

The invention discloses a BHK-21-SC cell strain adapted to serum-free suspension culture and a method for preparing a vaccine antigen with the cell strain. The BHK-21-SC cell strain is a juvenile golden hamster kidney cell strain, and has the preservation number of CGMCC No.16817. The cell strain is cultured according to the initial cell density of 0.5-0.6*10<6> cell / ml. 72 hours later, the cellsgrow to the cell density of 8-10*10<6> cell / ml. The screened and domesticated BHK-21 cell strain adapted to serum-free full suspension culture is used for production of a high-titer Newcastle diseasevirus solution, and a domestication process for preparation of Newcastle disease virus by the BHK-21 cell strain adapted to serum-free suspension culture is established.

The invention discloses a method for preparing virus of porcine reproductive and respiratory syndrome on a large scale. In the method, the virus of the porcine reproductive and respiratory syndrome is prepared in a cell microcarrier suspension culture system by a bioreactor. The method comprises the following steps of: inoculating host cells for preparing the virus to a carrier tank containing culture solution and a microcarrier, and mixing the cells and the microcarrier uniformly to ensure that the cells are attached to the microcarrier; providing sufficient nutrients and appropriate gas environment for the cells under the appropriate culture environment to ensure that the cells are grown until the cells are in an amount which are 10 to 20 times of the inoculation concentration on the microcarrier; preparing virus suspension from the virus of the porcine reproductive and respiratory syndrome by using cell maintenance culture solution to ensure that the suspension is adsorbed to the cells; culturing the virus under the appropriate culture environment; culturing continuously for 2 to 3 days to obtain virus solution; and after the virus solution passes inspection, performing freeze thawing on the virus solution twice at the temperature of -20 DEG C, and inactivating and purifying to prepare an inactivated vaccine of the porcine reproductive and respiratory syndrome or adding a freeze-drying protective agent for freeze drying to prepare a live vaccine of the porcine reproductive and respiratory syndrome. The method has large production scale, high yield of single batch and low production cost.

The invention discloses a production process of a porcine reproductive and respiratory syndrome virus (PRRSV) by using a bioreactor to culture a Marc-145 cell, which comprises the following technical steps of: (1) selecting a Marc-145 cell as a vaccine preparation cell; (2) culturing the vaccine preparation cell through passage and amplification; (3) reproducing seed virus for production; (4) culturing the microcarrier of the Marc-145 cell in the bioreactor; (5) producing an antigen of the PRRSV; and (6) treating the harvested virus antigen liquid. The invention can greatly reduce production cost and increase the input-output ratio by 5-10 times. Moreover, the invention has the advantages of short production period, small occupied land, easy and rapid expansion of production scale, less environment pollution, easy treatment, high automation degree, fewer people and easy realization of balanced and stable quality. The production cost can be obviously increased, and the yield and the quality of the vaccine can be increased.

The invention discloses a recombinant rabies virus rHEP-CaIL6 carrying an immune enhancement factor interleukin (IL) 6 gene and application thereof. The recombinant virus takes rabies virus HEP-Flury strain as the skeleton, the IL6 gene of canine is inserted to a position between the G and L gene of HEP-Flury to obtain a recombinant plasmid pHEP-CaIL6, and finally saving screening is carried out to obtain the recombinant rabies virus strain rHEP-CaIL6. The recombinant virus carries the immune enhancement factor, can enhance the immune response and induce the production of higher rabies virus neutralizing antibody, thus better protecting the body from resisting the attack of lethal rabies virus, also can produce a neutralizing antibody with protective ability at a low dose, and lowers the cost of canine vaccines. Moreover, the IL6 gene is recombined into the rabies virus, thus achieving stable expression of IL6 protein, also avoiding the overexpression thereof, and overcoming the defect that excessive IL6 can cause pathological injury.

The invention discloses a CA-193 virus strain (Coxackievirus A16) and application thereof to preparation of an inactivated vaccine. For the application, MRC-5 cells are used for proliferating the CA-193 virus strain; virus proliferation liquid is subjected to centrifugation, concentration and column chromatography filtration and purification after inactivation by formaldehyde or beta-propiolactone; and a CA16 inactivated vaccine is obtained. The invention provides the vaccine virus strain capable of being used for preparing a human CA16 inactivated vaccine; the subtype of the vaccine virus strain belongs to the main epidemiological subtype B1 in Chinese Mainland; the vaccine virus strain has good growth characteristics on the MCR-5 cells; and the virus titer (7.5 to 8.251gTCID50 / ml) with high stability can be obtained. A preparation method of the vaccine is complete; the impurity content is low; the immunogenicity and the immunizing protection performance are good; meanwhile, cell culture substrates are MRC-5 cells; the safety is high; and the international and CFDA specifications and requirements on the vaccine research and development can be well met.

The invention provides a preparation method for serum-free and suspension culture of HEK-293 cells and an application of the serum-free and suspension culture of the HEK-293 cells, and relates to the preparation method for the serum-free and suspension culture of the HEK-293 cells, a cell line obtained by the preparation method and the application of the cell line. The preparation method is few in passage times and high in domestication efficiency, and the obtained cells are high in viability, good in dispersity and stable in growth state and can meet the requirements of commercialized and large-scale production.

Immunization platforms, immunization regimes and medicaments useful for inducing an immune response in a mammal and preventing or treating a pathogenic infection in a mammal, wherein said immunization platforms and medicaments comprise a recombinant vesicular stomatitis virus (VSV) of one serotype and a rVSV of another serotype and are used in a prime-boost immunization regime. In aspects of the invention one VSV serotype is Indiana and the other VSV serotype is New Jersey.

The invention discloses a method for producing PRRS (Porcine Reproductive and Respiratory Syndrome) viruses through culturing Marc-145 cells by applying a torrent-perfusion bioreactor. The method comprises the following steps of: (1) selecting Marc-145 cells as cells for culturing seedlings; (2) subculturing the Marc-145 cells; (3) reproducing virus seeds for production; (4) perfusing and culturing the Marc-145 cells on a paper carrier in the torrent-perfusion bioreactor; (5) producing the PRRS viruses in the torrent-perfusion bioreactor; and (6) treating the obtained virus antigen solution. The invention can greatly decrease the production cost, improve the input-output ratio by more than 10 times and expand the production scale easily and rapidly, has short production preparation cycle and high degree of automation, occupies less area, causes less environmental pollution which is easy to treat, consumes less labor, is easy to realize balanced and stable virus quality and can obviously improve the yield and the quality of the viruses.

The invention discloses a production process of a porcine reproductive and respiratory syndrome virus (PRRSV) by using a bioreactor to culture a Marc-145 cell, which comprises the following technical steps of: (1) selecting a Marc-145 cell as a vaccine preparation cell; (2) culturing the vaccine preparation cell through passage and amplification; (3) reproducing seed virus for production; (4) culturing the microcarrier of the Marc-145 cell in the bioreactor; (5) producing an antigen of the PRRSV; and (6) treating the harvested virus antigen liquid. The invention can greatly reduce production cost and increase the input-output ratio by 5-10 times. Moreover, the invention has the advantages of short production period, small occupied land, easy and rapid expansion of production scale, less environment pollution, easy treatment, high automation degree, fewer people and easy realization of balanced and stable quality. The production cost can be obviously increased, and the yield and the quality of the vaccine can be increased.

The invention discloses a technology for the production of porcine epidemic diarrhea virus by means of the microcarrier culture of VREO cells using a bioreactor, and comprises the technology for the production of different porcine epidemic diarrhea virus strains. The technology comprises the following technical steps: (1) selection of VERO cells as cell line for vaccine; (2) passage and culture of cells for vaccine; (3) propagation of seed culture of the porcine epidemic diarrhea virus; (4) microcarrier suspension culture of the VERO cells in the bioreactor; (5) propagation of porcine epidemic diarrhea virus antigen; and (6) treatment of acquired virus antigen liquid. The production method can remarkably lower production cost and enhance output-input ratio by 5 to 10 times, and has the advantages of short production period, small occupied space, great easiness for enlarging production scale rapidly, little environmental pollution, easy processing, high automation degree, a small number of staff, easy implementation of even and stable quality, obviously lowered production cost and enhanced yield and quality of vaccine.