79results about How to "High virus content" patented technology

The invention relates to a super-sensitivity method, primer group and kit for capturing whole genome of novel coronavirus. The method can conveniently and quickly amplify the whole genome of the novelcoronavirus by using a small amount of RNA, and can directly dock with a second-generation sequencing library-building reagent and a third-generation sequencing platform so as to obtain a whole genome sequence of the novel coronavirus.

The invention provides a method of preparing a vaccine through suspension culture of mammal cells, which includes the following steps: (1) preparing microencapsulated animal cells through a microencapsulation method; (2) inoculating the microencapsulated animal cells in a cell culture system to perform cultivation; and (3) inoculating virus into the cell culture system to culture the virus, and obtaining a virus liquid to prepare the vaccine. By means of the microencapsulation method to culture the animal cells, a problem of large-scale continuous amplification of cells is solved. When the cultured animal cells are used for breeding virus, a high content of virus is achieved, so that the virus liquid, when being used for preparing the viral vaccine, can also achieve a better immune effect. The invention also provides a method of preparing a cell-expression product through the suspension culture of the mammal cells. The method reduces dependency on micro-carriers and is greatly reduced in production cost.

The invention provides a combined live vaccine for preventing porcine reproductive and respiratory syndrome and pseudorabies, and a preparation method and application thereof. According to the invention, no immunosuppression occurs between two vaccines of the combined live vaccine; compared with each single vaccine, the combined live vaccine has no obvious difference in security, immunogenicity, immunity duration and immuno-protective effects and has remarkable immuno-protective effects on preventing porcine reproductive and respiratory syndrome and pseudorabies.

The invention discloses a method for mass production of pseudorabies virus vaccine, comprising the following steps: (a) adding netty polyester fiber which serves as a carrier into a bioreactor provided with a tide type micro-carrier suspension culture system and inoculating cells for producing vaccine; (b) inoculating pseudorabies virus vaccine when the culture cell grows to a certain intensity, so that the cells are infected by the pseudorabies virus vaccine; (c) reproducing the virus in great numbers under appropriate conditions; (d) harvesting the virus when cytopathic rate reaches above 70%; (e) carrying out freeze thawing on the harvested virus for once or twice to lead the cells to completely come off and disperse and then adding freeze-drying protective agent, evenly mixing the mixture, packaging the mixture in fixed volume and freeze-drying.The method of the invention has the advantages of good stability, explicit process control indicators, good controllability, easy operation, large process scale and the like.

The invention relates to a method for producing porcine circovirus type 2 antigens in large scale with high density. A bioreactor microcarrier suspension culture technology used for replacing an existing spinner bottle culture technology for producing the porcine circovirus type 2 antigens. The method can greatly reduce the production cost and improve the yield. Compared with the spinner bottle technology, the unit antigen cost is reduced by 80% to 90%, the production period is reduced by 7 days, and the yield is improved by 3 times to 10 times; due to the fact that the antigens can be obtained repeatedly, compared with a traditional reactor culture technology, the unit antigen cost is reduced by 40% to 50%, and the yield is improved by 3 times to 5 times. The antigens produced through the method has no serum residues, the produced vaccine is higher in safety and small in batch difference, the quality is stable and easy to control, and the yield and quality of the produced vaccine can be obviously improved.

The invention discloses a method for producing duck tembusu virus inactivated vaccines in a large scale. According to the method, high-density culture of cells is performed by using a bioreactor culture technology to produce the duck tembusu virus inactivated vaccines. Compared with a conventional rotating bottle production process, the method for producing duck tembusu virus inactivated vaccines in a large scale has the characteristics that the automated control degree is high, and the production can be monitored in real time to ensure that the product quality is uniform and stable; and manpower and material resources can be saved during production to reduce the production cost; and the produced venom is high in virus content and low in inter-batch difference, and side effects are reduced.

The invention relates to a method for establishing a hog cholera lapinized virus labeled vaccine strain and preparing a vaccine. The method comprises the following steps: (1) establishing the overall-length infectious clone of the hog cholera lapinized virus strain (the C strain or the Chinese strain); (2) introducing labeled protein genes; (3) rescuing the hog cholera lapinized labeled vaccine virus; (4) breeding the virus; (5) inspecting a virus solution; (6) preparing a vaccine; and (7) inspecting the finished product of the vaccine, comprising safety inspection and efficacy inspection. The hog cholera lapinized virus labeled vaccine established by the invention maintains the characteristics of good safety, excellent immunogenicity and the like of the hog cholera lapinized virus strain; the virus strain has the advantages of good stability and high cell production virus content, can be used for industrial mass production and can generate a labeled protein specificity antibody aftera hog is immunized to distinguish immunization and natural infection and has important significances on hog cholera prevention, control and purification and emergent immunization.

The invention relates to a marking vaccine for the porcine reproductive and respiratory syndrome high in pathogenicity and a preparation method thereof.A virus seed produced through the vaccine is a constructed JXA1-RM strain, safety and immunogenicity of the JXA1-RM strain are maintained, in addition, the strain is good in stability, and the content of viruses produced by cells is high; after a pig is immunized through the vaccine, no sequence antibody against the M protein linear list position peptide amino acid sequence of the PRRS virus of an American strain is generated, and the vaccine can be used for differentiating an immunized animal from a naturally-infected animal.By means of a method for identifying and diagnosing the strain, antibodies generated by a field strain and a vaccine strain can be diagnosed, a conventional ELISA detection method is combined, an animal immunized through the vaccine and a naturally-infected animal can be differentiated, and it is beneficial to purification of the porcine reproductive and respiratory syndrome high in pathogenicity.

The invention provides a Muscovy duck parvovirus and gosling plague bivalent vaccine. The antigens used by the vaccine is inactivated Muscovy duck parvoviruses and Muscovy duck-source gosling plague viruses, the preservation number of the Muscovy duck parvoviruses is CGMCC No. 8504, and the preservation number of the Muscovy duck-source gosling plague viruses is CCTCC No. V201620. A preparation method of the Muscovy duck parvovirus and gosling plague bivalent vaccine includes: the Muscovy duck parvovirus YBMDP strains and Muscovy duck-source gosling plague virus YBGPV-M strains which are high in virus content and good in immunogenicity are screened, infected embryos and allantoic fluid are collected after duck embryo inoculation, and oil emulsion adjuvant is added for emulsification and mixing to obtain the vaccine after homogenization, ultrafiltration and concentration, and formaldehyde solution inactivation. The prepared vaccine can immunize breeding Muscovy ducks and increase the level of two types of antibodies of the breeding Muscovy ducks at the same time, guarantee the offspring maternal antibody level of the breeding Muscovy ducks, and prevent the young Muscovy duck parvovirus diseases caused by the Muscovy duck parvoviruses and gosling plague virus infection caused by the Muscovy duck-source gosling plague viruses.

The invention belongs to the technical field of veterinary biological products, and particularly relates to a preparation method of a duck reovirus inactivated vaccine. In the invention, an LMH passage cell line is used as a carrier cell for virus proliferation; a culture medium A consisting of lysine, recombinant human insulin, human serum albumin, protamine, biotin, polysaccharide sulfate and agrowth factor is used for culture; and the virus fluid is collected and inactivated to prepare a vaccine. The LMH passage cell line is used as a carrier cell for virus proliferation without the need of adding pancreatin, and the LMH cells have a clear background, have no exogenous pathogens and are easy for proliferation, which can effectively simplify the process and reduce costs. Meanwhile, theduck reovirus inactivated vaccine prepared by the invention has high virus content, good stability and high safety, and is an ideal duck reovirus inactivated vaccine.

The invention provides a porcine reproductive and respiratory syndrome virus liposome diluent lyophilized product and a preparing method thereof, and belongs to the technical field of animal biological products. 1000 ml of a PBS solution contains 2-8 g of astragaloside III, 5-10 g of aloe polysaccharide, 2-6 g of curcumin, 10-15 g of lipidosome, 10-15 g of glucose and 10-15 g of mannitol. The prepared liposome diluent lyophilized product can reduce difference between semi-finished product batches and can promote propagation of porcine reproductive and respiratory syndrome viruses, increase virus content and enhance immunogenicity of vaccines.

The invention discloses a porcine circovirus 2-type lipidosome diluent lyophilized product and a preparation method thereof, and belongs to the technical field of animal biological products. Each 1,000 ml of PBS solution is prepared from 5 g to 10 g of cordyceps polysaccharide, 5 g to 10 g of lycopene, 1 g to 5 g of ginsenoside Rh2, 6 g to 12 g of lipidosome, 10 g to 15 g of saccharose and 10 g to 15 g of mannitol. According to the prepared lipidosome diluent lyophilized product, the difference between batches of semi-finished products can be reduced. The prepared lipidosome diluent lyophilized product can promote reproduction of porcine circovirus 2 type, the virus content can be increased, and vaccine immunogenicity can be enhanced.

The invention provides a method for preparing inactivated rabies vaccine for animal by a bioreactor. The method includes by the aid of the bioreactor and a Cytodex-1 microcarrier system, subjecting high-density Vero cells (or BHK cells) adaptive to rabies virus CTN-1 strain (aG-strain or PV-strain) to virus infection by utilizing the CTN-1 strain (aG-strain or PV-strain) rabies virus with good antigenicity so as to prolong virus harvest time from 6-9 days to 11-15 days, increase virus titer from 105-7LD50/ml to 108-9LD50/ml and increase virus content by 10-1000 times; and inactivating harvest virus liquid, adding additives, and then packing to obtain high-quality vaccine products. The prepared vaccine is safe and effective, production process is stable, and production quality is controllable.

The invention provides a duck plague live vaccine and a preparation method thereof, and belongs to the technical field of biological product preparation. A chicken embryo fibroblast line DF-1 is takenas the host of duck plague viruses to produce a duck plague virus solution, and then a heatproof protective agent is added into the duck plague virus solution to prepare the duck plague live vaccine.DF-1 is used to culture duck plague viruses to avoid the safety hazard that the duck plague viruses cultured by (chicken embryo fibroblast) CEF cells are contaminated by exogenous viruses; the DF-1 cells are available at any time, time and labor are saved, and the production cost is greatly reduced. The prepared duck plague live vaccine has the advantages of good safety and high immunity effect,and can protect ducks from strong duck plague viruses. Furthermore, the live vaccine also comprises an improved heatproof protecting agent, thus the duck plague live vaccine can be stored for 24 months at a temperature of 2-8 DEG C, the virus content an titer are not reduced, the storage conditions are lowered, the storage and transportation of the live vaccine become convenient, and the production cost is reduced.

The invention discloses a recombinant gene VII type Newcastle disease virus attenuated strain rN7a strain, and further discloses a vaccine composition containing the rN7a strain or a culture-inactivated antigen of the rN7a strain. The rN7a strain is the attenuated strain obtained by replacing a P protein gene sequence of a Newcastle disease virus N7a strain with the preservation number of CCTCC NO:V201545 with a P protein gene sequence as shown in SEQ ID No.1. The rN7a strain has high virus titer and high HA titer after culture. The vaccine composition can provide complete protection to a variety of strains.

The invention provides a method for preparing pseudorabies live vaccines from DF1 continuous cells and a product prepared by the method. The method includes the steps of culturing a pseudorabies virus attenuated strain by the DF1 continuous cells; obtaining cell culture virus fluid; adding stabilizers and antibiotics and obtaining the pseudorabies DF1 live vaccines after vacuum freeze drying. The method has the advantages that the DF1 continuous cells which are heterogenous continuous cells are used for culturing pseudorabies attenuated viruses, so that the possibility that homologous cells have unknown swine-source microorganisms can be avoided and pure vaccines can be better guaranteed.

The invention provides a method for purifying a foot-and-mouth disease virus antigen by utilizing ion-exchange chromatography, which comprises the following steps: inactivating and filtering a harvested foot-and-mouth disease virus solution, and sequentially carrying out PEG (Polyethylene Glycol) precipitation, redissolution, ion-exchange chromatography and the like. Wherein the ion exchange chromatography comprises the following steps: adopting a flow-through process, enabling a redissolved solution to pass through a chromatographic column, and collecting a flow-through component. By adopting the process, the yield of three subtype inactivated antigens of foot-and-mouth disease AKT-III, O/Mya 98 and OHM02 is more than 90%, the purity is more than 70%, the content of impure protein in the antigens is greatly reduced, more than 50% of total protein can be removed, the process is stable in amplification and short in time consumption, the process can be used for preparing high-quality foot-and-mouth disease vaccines, and the safety and effectiveness of the foot-and-mouth disease vaccines are improved.

The invention discloses a veterinary bovine ephemeral fever inactivated vaccine and a large-scale production method thereof, and belongs to the technical field of vaccine production. The method provided by the invention comprises the steps of performing cell culture by utilizing a suspension culture process, performing virus suspension culture, performing virus antigen solution clarification, performing concentration and purification, performing inactivation and performing emulsification vaccine preparation, and the method can realize mass proliferation of the bovine ephemeral fever virus, improves the content and yield of the virus antigen, and has a high automation degree; The synergistic effect of all the steps can jointly improve the quality and stability of the vaccine, and the technical problems that the yield of the bovine ephemeral fever virus produced by cell culture through a traditional spinner bottle or microcarrier process is low, the process is tedious, the difference between virus antigen batches is large, the vaccine efficacy is low, the vaccine validity period is short and the like are solved.