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

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 relates to a recombinant vaccine strain for foot-and-mouth disease type A, which is constructed by a gene recombination technology and featured with high valence, high antigen compatibility and high immune protective rate, as well as a preparation method thereof and an application thereof. An antigen nucleotide sequence of the vaccine strain is shown by SEQ ID NO. 1; a rescuing system is an efficient eukaryotic plasmid, which is manually constructed and can express an accurate foot-and-mouth disease virus genome RNA; therefore, a foot-and-mouth disease recombinant virus can be constructed and prepared; through the adoption of the plasmid, the vaccine strain with high titer and good antigen compatibility can be prepared so as to prepare inactivated vaccines; after pigs and cattle are immunized, bodies can be effectively stimulated to generate immune responses and immune protective effects on the pig and cattle bodies can be provided; an A type AISA lineage of strain is challenged with 10000 times of 50% cattle infective dose (BID50), so that the immune protective rate reaches 100% and 50% protective dose (PD50) is 10.81-13.59; and the vaccine can be used for preventing and controlling foot-and-mouth diseases type A in China and neighboring countries.

The invention relates to a VII type Newcastle disease virus L-gene mutation attenuated vaccine strain produced by using a reverse genetic manipulation technology and a preparation method of the VII type Newcastle disease virus L-gene mutation attenuated vaccine strain, and belongs to the field of biological products and biological technologies. Toxicity of the VII type Newcastle disease virus L-gene mutation attenuated vaccine strain is weakened obviously, and at the same time, the vaccine strain on a chick embryo has high titer, stable heritability and high compatibility to an antigen of a prevalent virus, is suitable for large-scale production of vaccines, and can be used for producing the vaccines. Compared with conventional vaccines (gene I and gene II types), the VII type Newcastle disease virus L-gene mutation attenuated vaccine strain has a wide application prospect in control of incidence and prevalence of the Newcastle diseases.

The invention relates to a Seneca Valley virus recombinant plasmid and a Seneca Valley virus recombinant virus, and construction methods thereof, and belongs to the technical field of virus construction. The Seneca Valley virus recombinant virus is a Seneca Valley virus with the 177th amino acid, mutated to A from S, of a GH ring of capsid VP2. The Seneca Valley virus recombinant virus has a highvirus tilter and a high virus proliferation ability.

The invention relates to a SARS-CoV-2 inactivated vaccine and a preparation method of the vaccine. The preparation method comprises the following steps: step (1) recovery and large-scale culture of Vero cells; step (2) inoculation of working seeds, batch of virus seeds, virus culture and one-time harvest virus liquid, namely, the virus harvest liquid; step (3) obtaining an inactivated virus concentrated solution after a first virus inactivation, concentration and a second virus inactivation; step (4) obtaining a vaccine stock solution after purifying the inactivated virus concentrated solution; and step (5) preparation of semi-finished product and sub-package after the stock solution is verified to be qualified.

The invention discloses a recombinant rabies virus rHEP-Flury(dG-VP2) carrying two genes G and a gene VP2. According to the recombinant virus, a rabies virus HEP-Flury strain serves as a framework, the VP2 gene shown as SEQ ID NO.1 is inserted into HEP-Flury, an additional gene G of the rabies virus is inserted, and the recombinant plasmid pHEP-Flury(dG-VP2) carrying the two genes G and the gene VP2 is obtained; finally, rescuing screening is carried out, and the recombinant rabies virus rHEP-Flury(dG-VP2) is obtained. The recombinant rabies virus rHEP-Flury(dG-VP2) carries the two genes G and expresses VP2 protein, a high rabies virus resisting antibody and canine parvovirus VP2 resisting antibody level can be generated through immune induction, the cost of vaccines for dogs can be reduced, and very good application and popularization prospects are achieved.

The invention discloses a human-derived rotavirus attenuated strain with wide-spectrum immunogenicity and a vaccine prepared from the human-derived rotavirus attenuated strain. The human-derived rotavirus attenuated strain with wide-spectrum immunogenicity is named as HRV-LZ-2013, has a gene type of G10P8, is preserved in the China general microbiological culture collection center and has a strain preservation number of CGMCC N0. 9452. An experiment proves that the rotavirus attenuated strain keeps low pathogenicity to animals. An immunization protection experiment on animals and especially on rotavirus-caused diarrhoea model rabbit and pig proves that the attenuated live vaccine or inactivated vaccine prepared from the human-derived rotavirus attenuated strain has good immunizing protection properties. An immunizing protection test on cattle and sheep proves that the vaccine provided by the invention has a wide immunizing protection range and can be used for preventing cattle, pig, horse, sheep and dog diarrhoea caused by rotaviruses.

The present invention relates to herpes virus family medical microorganism. The present invention is prepared by the following method: (1) serial passage MRC-5 cell; (2) inoculating to cell and continuous infecting; (3) adding culture solution; (4) washing cell surface; (5) eluting infected cell, centrifugal separating, adding vaccine fluid containing stabilizing agent to refrigerate; (6) ultrasonic crashing and centrifugal separating to acquire virus stock solution; (7) combining and diluting, and at once processing to pack and freeze-dry, preparing attenuated pox vaccine. The stabilizing agent is prepared by dissolving NaCl, KCl, KH2PO4, Na2HPO4.12H2O, saccharose, sodium glutamate, kalamycin, erythromycin and deionized water into the water with weight ratio 3:0.1:0.2:3.3:50:0.4:0.1:0.03:1000. The present invention has good advantages of safety and immunogenicity.

The invention provides an infectious bursal disease virus Vero cell-adapted strain and belongs to the field of bioengineering. The related infectious bursal disease virus Vero cell-adapted strain is named Ck /Jiangsu/ NJ-23/2008 and has a collection No. of CGMCCNO.8852. The infectious bursal disease virus Vero cell-adapted strain which can efficiently multiply on serum-free cultured Vero cell is finally obtained through wild strain separation, chick embryo passage, Vero cell passage adaption; the infectious bursal disease virus Vero cell-adapted strain is subjected to continuous passage culture on the serum-free cultured Vero cell and TCID50 can be kept to be higher than 108.5/mL. Virus culture solution is inactivated and prepared into oil emulsion; after the prepared oil emulsion is used to immunize chicken, detection proves that the prepared oil emulsion has good immunogenicity. The infectious bursal disease virus (IBDV) strain and the production process thereof are simple, safe and efficient and suitable for industrial culture.

The invention relates to the technical field of biology and discloses sgRNA of ID1 (Inhibitor of Differentiation 1) genes, a knock-out method for ID1 genes, a BHK-21 cell line and the application thereof. Sequences of the sgRNA are shown as any pair of SEQ ID NO:1-2 or SEQ ID NO:3-4. Because the ID1 is capable of inhibiting foot and mouth disease virus multiplication, by knocking-out the ID1 genesin BHK-21 cells, the virus titer of an ID1 knock-out cell line is increased, so that the production cost of the foot and mouth disease virus is reduced.

The invention discloses a large-scale production method for veterinary pseudorabies attenuated live vaccines. The method comprises the steps of culturing pseudorabies virus attenuated strains by using a cell suspension process so that the density of BHK21-C13 cells subjected to suspension culture in a reactor is more than or equal to 2*10<6> cells per milliliter, then culturing with a virus maintenance solution containing pseudorabies suspended virus seeds, and performing clarification, seedling, packaging and freeze-drying on the harvested pseudorabies attenuated virus solution to obtain the veterinary pseudorabies attenuated live vaccines. By adopting the method, mass multiplication of pseudorabies attenuated viruses can be realized, the titer of viruses and the yield of antigen are greatly improved, process automation is realized, the quality of the vaccines is improved, the density of the BHK21-C13 cells can be improved, the optimal biochemical condition of cell growth or virus multiplication can be automatically monitored, the titer of viruses is improved, large-scale production of the process is relatively easy, and the vaccines are stable in quality and low in batch difference and have broad production and application prospect.

The invention discloses a recombinant rabies virus carrying a deoptimized M gene and two G genes. Firstly, a reading frame part of the M gene is deoptimized, wherein the sequence after deoptimization is as shown in SEQ ID NO.2; secondly, a rabies virus HEP-Flury strain is taken as a skeleton, an M gene in the HEP-Flury is replaced by the deoptimized M gene, and an additional rabies virus G gene is inserted to obtain recombinant rabies virus pHEP-dG-Mmin plasmid carrying the deoptimized M gene and two G genes; finally, rescuing and screening are conducted to obtain a recombinant rabies virus strain rHEP-dG-Mmin. The recombinant virus has higher virus titer, and the cost of canine vaccine can be reduced. Under the condition of multiplicity of infection, G protein with higher level can be expressed, virus replication and transcription of each structural gene are increased, and the recombinant virus has the potential of serving as a rabies vaccine candidate strain.

The invention discloses a production process for producing porcine circovirus type 2 by culturing cells pK-15A1# in a bioreactor. The production process comprises the technical steps of (1) selecting pK-15A1# cells for multiplying circovirus type 2; (2) utilizing the bioreactor as a cell culture tool; (3) carrying out a virus culture process of synchronous virus inoculation; (4) culturing the pK-15A1# cells on micro-carriers in the bioreactor; (5) carrying out multiplication culture on viruses of porcine circovirus type 2 in the cells on the micro-carriers in the bioreactor; and (6) harvesting virus culturing materials of porcine circovirus type 2. The production process has the advantages that the production cost can be lowered, and compared with that of a traditional spinner bottle production process, the input-output ratio of the production process is increased by 5-10 times; the production cycle is short, the occupied area is small, the production scale can be easily and rapidly expanded, the environmental pollution is slight and is easy to avoid, the automation degree is high, few workers are required, the quality is stable, the cost can be remarkably lowered, and the yield and quality of vaccines can be remarkably improved.

The invention discloses a feline herpesvirus I type virus strain and application thereof, and belongs to the technical field of biological products. The preservation number of the virus strain is CCTCC NO: V202126, the preservation time is April 27, 2021, and the preservation address is Wuhan University, Wuhan, China. The virus strain belongs to an isolated strain in Wuhan of China, has high virus titer of 107.65 TCID50/mL, has strong pathogenicity to domestic cats and has the basic potential of becoming a vaccine candidate strain, a vaccine prepared from the virus strain can be used for preventing and treating feline infectious rhinotracheitis diseases, an important vaccine virus source is provided for prevention and control of the feline herpesvirus I in China, and effective prevention and control of the feline herpesvirus I are achieved.

The invention provides a porcine epidemic diarrhea virus as well as a separation method and application thereof, relating to the technical field of biology. According to a separation purification method of the porcine epidemic diarrhea virus, the porcine epidemic diarrhea virus is separated by virtue of polyethylene glycol 6000. The method is simple in operation and strong in practicability, the porcine epidemic diarrhea virus can be effectively separated, and the separated porcine epidemic diarrhea virus has relatively high purity and virus titer. A tissue inactivated vaccine prepared from the porcine epidemic diarrhea virus has the beneficial effects that impure proteins are few, the batch difference of the vaccines is small, the immune effect is good, and the side effect is slight.

The invention relates to a BHK cell line stably expressing a hamster TIGAR gene, and a construction method and an application thereof. The cell line contains the TIGAR gene and has the preservation number of CCTCC NO:C201928. The hamster TIGAR gene is amplified firstly, and a plasmid is constructed; then the constructed plasmid is transferred into 293T cells with PAX and PMD2G plasmids at the sametime to form packaged lentivirus; after the BHK cells are infected by the packaged lentivirus, drug screening is performed, and then enlarged cultivation is performed to obtain the cell line. The cell line can increase the anti-apoptotic ability of cells, prolong the survival time of the cells, increase the viral titer of Newcastle disease in the cells, and successfully construct and screen high-yield cell lines suitable for the reproduction of Newcastle disease virus.

The invention discloses a cell affinity agent capable of improving duck flavivirus in-vitro infection efficiency and a preparation method of the cell affinity agent and belongs to the technical field of agents for microbial virus culture. The cell affinity agent is prepared from the following components in percentage by mass: 0.5-5% of organic acid, 0-1% of phosphoric acid, 0-3.5% of inorganic salt and the balance of deionized water. By the cell affinity agent, the microenvironment between the duck flavivirus and the cell can be improved, and the affinity between the duck flavivirus and the cell is increased so that the infection efficiency of the duck flavivirus to the cell is effectively increased.