68 results about "Virus suspension" patented technology

Long rod shaped M13 viruses were used to fabricate one dimensional (1D) micro- and nanosized diameter fibers by mimic the spinning process of the silk spider. Liquid crystalline virus suspensions were extruded through the micrometer diameter capillary tubes in cross-linking solution (glutaraldehyde). Resulting fibers were tens of micrometers in diameter depending on the inner diameter of the capillary tip. AFM image verified that molecular long axis of the virus fibers were parallel to the fiber long axis. Although aqueous M13 virus suspension could not be spun by electrospinning, M13 viruses suspended in 1,1,1,3,3,3-hexafluoro-2-propanol were spun into fibers. After blending with highly water soluble polymer, polyvinyl 2-pyrolidone (PVP), M13 viruses was spun into continuous uniform virus blended PVP (virus-PVP) fibers. Resulting virus-PVP electrospun fibers showed intact infecting ability to bacterial hosts after suspending in the buffer solution.

The invention discloses a univalent and bivalent inactivated vaccine for preventing Enterovirus 71 (EV71) and Coxsackie virus A16 (Cox.A16) of hand-foot-and-mouth disease, and a preparation method thereof. The preparation method comprises the following steps: screening vaccine strains for the hand-foot-and-mouth disease; establishing and verifying a vaccine strain third-stage seed lot library; culturing cells; inoculating and propagating virus; collecting virus suspension; inactivating the virus; ultra-filtering, concentrating and purifying the virus suspension to obtain vaccine stock solution; and finally preparing the univalent and bivalent inactivated vaccine. The univalent and bivalent inactivated vaccine has good application prospect for preventing the hand-foot-and-mouth disease.

The invention discloses a univalent and bivalent gene engineered subunit vaccine for preventing Enterovirus 71 (EV71) and Coxsackie virus A16 (Cox.A16) of hand-foot-and-mouth disease, and a preparation method thereof. The preparation method comprises the following steps: respectively obtaining recombinant baculovirus Bac-EV71-P1-3CD and Bac-Cox.A16-P1-3CD by gene engineering means, respectively efficiently coexpressing similar SeQ ID No.1 EV71 P1 and Se Q ID No.2 Cox.A16 P1 and 3CD proteins in insect cells, and respectively self-assembling into EV71 VLP and Cox.A16 VLP; establishing and verifying a vaccine strain third-stage seed lot library; culturing cells; inoculating and propagating virus; lysing the cells, ultra-filtering and purifying virus suspension; and further preparing the univalent and bivalent vaccine. The vaccine has good application prospect for preventing the hand-foot-and-mouth disease.

The present invention relates to a composition comprising a viral antigen, a first protein and a second protein. Optionally, the composition also comprises three different disaccharaides, or, optionally, the composition comprises a primary sugar and at least one, preferably two secondary sugars. The present invention also relates to the use of a viral antigen, a first protein and a second protein for the manufacture of a composition, preferably a vaccine. The present invention furthermore relates to a method of treatment or prevention of virus associates diseases in humans. Moreover, the present invention relates to a method of adapting a virus to a suitable cell-line. The invention is also useful for the production of virus suspensions suitable for making stable, live/inactivated, monovalent and/or polyvalent, liquid/lyophilized rotavirus vaccine compositions for oral and/or nasal or any other suitable route of administration in human.

The invention discloses a TK (Thymidine Kinase) gene removed recombinant VTT (Tian Tan strain) oncolytic VACA (Vaccinia Virus). A VTT oncolytic VACA is subjected to recombination transformation, a TKgene is removed, and an EGFP (Enhanced Green Fluorescent Protein) gene and a Luciferase gene are expressed. The invention further discloses a preparation method of the TK gene removed recombinant VTToncolytic VACA, which comprises the steps of: recombining a wild type VTT virus and a targeting plasmid aiming at the TK gene of the VTT virus in cells to obtain virus suspension; screening and purifying the virus suspension to obtain the attenuated VTT oncolytic VACA. According to the TK gene removed recombinant VTT oncolytic VACA, which is disclosed by the invention, the TK gene is used as an attenuation target, the EGFP gene and the Luciferase gene are expressed, virus virulence is reduced, and tumor selectivity of the virus is improved; the TK gene removed recombinant VTT oncolytic VACA has a function of monitoring distribution of virus particles in a body, provides a novel antitumor drug for the clinic, can be used for researching and developing a live vaccine and is applicable to various tumors such as the lung cancer, the liver cancer, the melanoma and the like.

The invention provides a method for industrially producing porcine Japanese encephalitis (JE) vaccines by utilizing a bioreactor. The method comprises the following steps: (1) sterilizing a microcarrier and the bioreactor, then inoculating cells for vaccine preparation for culturing, inoculating JE viruses when dense monolayers are formed on the cells on the microcarrier, and continuing culturing to reproduce the viruses; (2) stopping culturing and harvesting virus suspension when cytopathy reaches more than 80%; (3) carrying out ultrafilter concentration as well as virus inactivation on the harvested virus suspension; and (4) purifying the inactivated viruses by adopting column chromatography to prepare inactivated vaccines. In the method, the technology of using the microcarrier in the bioreactor for culture is adopted to carry out high density culture of the cells to produce the porcine JE vaccines. Compared with the traditional spinner bottle production method, the method has the following advantages: the automatic control degree is high, so production can be monitored in real time; the labour is saved, thus reducing the cost; the land for production is few, thus being easy to enlarge the scale of production; and the produced viruses have high titer, so the batch-to-batch variation is small, the product quality is stable and the side reaction is low.

The preparation process of reovirus antigen subunit vaccine for grass carp includes the following steps: proliferation of GCRV873, which is the Hunan Shaoyang strain of reovirus, in grass carp kidney cell line; centrifugal purification and concentration of GCRV873 infected cell virus suspension; processing purified virus with surfactant to disintegrate complete virus structure to form viral nucleic acid and capsid protein subunit component; digesting with nuclease to degrade free virus genome dsRNA; dialysis to obtain reovirus protein subunit antigen preparation for grass carp containing no nuclease; and diluting with physiological saline to obtain the said vaccine. The vaccine of the present invention has the advantages of containing complete virus protein antigen component, containing no genetic virus matter RNA, simple preparation process, high product purity, etc.

The invention discloses a method of producing a foot-and-mouth disease vaccine by using a large-scale high-density BHK-21 cell adherent culture technology. The method comprises the following steps of: 1) thawing and primarily amplifying cells; 2) conducting large-scale and high-density cell culture in a medium-volume bioreactor, and after the culture is completed, diluting the high-density cell sap in a large cell culture pot; and 3) inoculating foot-and-mouth disease seed virus, harvesting all virus suspension under proper conditions, and preparing the foot-and-mouth disease vaccine through the following steps. The process method is free from the process of acclimating the adherent cells into suspension cells; the necessarily gradual amplification process in the existing technology of preparing the foot-and-mouth disease vaccine by conducting large-scale suspension culture of the BHK-21 cell can be avoided, the technology flow can be obviously simplified, the production cycle can be shortened, the escape of the foot-and-mouth disease virus can be furthest reduced, and the pollution possibility can be decreased.

The invention provides a method for preparing genotype F mumps attenuated live vaccine. The method comprises the following steps of inoculating mumps attenuated strain MHM-19 to human embryo lung diploid cell MHL-3 which is cultured to a monolayer; releasing virus through frozen-thawed cell in the peak of viral multiplication; centrifuging and removing cell debris; adding a protective agent to centrifuged virus suspension to obtain a semi-finished product of vaccine; and freezing and drying the semi-finished product to obtain the finished product of genotype F mumps attenuated live vaccine. After a rhesus monkey is injected with the prepared MHM-19 attenuated live vaccine for immunizing, good humoral immunity is induced to be produced, and the detection results obtained at different times after immunization show that the MHM-19 attenuated live vaccine has immunizing potency superior to that of reference mumps vaccine S79; in addition, the safety experiment shows that the rhesus monkey does not suffer from parotitis-related symptom after being injected with MHM-19 attenuated live vaccine, the pathological tissue nearly has no obvious difference with that of S79 strain by observing, so that the MHM-19 attenuated live vaccine has high safety.

The invention mainly aims to provide an ectropis oblique warren nuclear polyhedrosis virus insecticidal suspending agent and a production method thereof. The insecticide consists of virus suspension, silicon dioxide, aromatic amino acids, a refined fluorescent whitening agent, sodium lignin sulfonate, polyvinyl alcohol, chlorfluazuron, glycerin and sterile water. The production method comprises the following steps: paired spawning on pupas by virtue of emerged adults, sterilizing the eggs, hatching larvae, feeding the larvae by using artificial feed, inoculating ectropis oblique warren virus on the feed surface to feed, grinding and filtering the larvae infected with the virus, centrifuging filtrate, collecting precipitates, and preparing the corresponding insecticide. By utilizing the insecticide, plant diseases and insect pests of ectropis oblique warren can be effectively controlled, the control cost is low, the environment friendliness is promoted, and the problems that the drug resistance of injurious insects is enhanced, the natural enemy number is reduced and pesticide residues in tea leaves are increased due to use of lots of chemical pesticides are solved.

The invention relates to the field of virus suspension culture and provides a method for culture of Newcastle disease virus by suspension cells. The invention provides application of a DMEM culture medium to increasing of cell agglomeration rate in a process of virus culture with the suspension cells. The invention further provides a method for culturing gene VII-type Newcastle disease virus by the suspension cells. The method is characterized by including: culturing AGE1 cells until density reaches (8-12)*10<6>/ml, performing inoculation of the Newcastle disease virus, adding a serum-free DMEM culture medium accounting for 10%-20% of the volume of a cell culture system, daily adding pancreatic enzymes in final concentration of 8-16U/ml, culturing at a temperature of 33-37 DEG C, and collecting the virus. The AGE1 is used for culturing the gene VII-type Newcastle disease virus for the first time, and by adoption of the serum-free DMEM culture medium and other reasonable process parameters, high-titer Newcastle disease virus can be obtained.

The invention belongs to the technical field of veterinary biological products and particularly relates to a pseudorabies/porcine parvovirus infection combined inactivate vaccine and a suspension culture preparation method. The preparation method comprises the following steps: preparing a pseudorabies virus suspension culture antigen and a porcine parvovirus infection virus suspension culture antigen; proportionally mixing the inactivated pseudorabies virus and porcine parvovirus infection virus antigen solutions, and then adding an adjuvant to fully emulsify to obtain the pseudorabies/porcineparvovirus infection combined inactivate vaccine. The suspension culture processes of the pseudorabies virus antigen solution and the porcine parvovirus infection virus antigen solution are established, the virus antigen solutions prepared through the suspension culture process have the advantages of high antigen content, large batch of the antigen and stable batch, the preparation method greatlyreduces the use of manpower, the occupied area and space of a culture system is reduced, and the production cost of an enterprise is lowered; meanwhile, the pseudorabies/porcine parvovirus infectioncombined inactivate vaccine is used, two viruses are prevented through one syringe, the immunity times of an animal are reduced, the stress times of the animal are reduced, and the production cost ofthe vaccine and the culture cost of a farmer are greatly reduced.

The invention relates to a method for purifying a slow virus. The method comprises the following steps: (S10) providing a cell culture containing the slow virus; (S20) carrying out centrifugal concentration on the cell culture, and filtering to obtain virus suspension; (S30) purifying the virus suspension by virtue of anion exchange chromatography; and (S40) purifying the virus suspension by virtue of gel filtration chromatography. A process for purifying the slow virus is easy to amplify, can be used for processing large amounts of virus liquid produced by cell factories or bioreactors on a large scale, can meet production requirements of slow virus liquid, is stable and is good in repeatability; furthermore, the purified slow virus liquid prepared by virtue of the method is not polluted by exogenous factors, is low in residues of cell host protein and host DNA, good in safety, and high in purity and titer and can completely meet requirements of clinic gene therapy.

The invention provides a buzurasuppressarianucleopolydro virus and beauveria bassiana insecticidal suspending agent, and a preparation method thereof. The effective components comprise a uzurasuppressarianucleopolydro virus suspension solution and beauveria bassiana powder at a weight ratio of 1:12-2:1. The buzurasuppressarianucleopolydro virus and beauveria bassiana insecticidal suspending agentpossess synergistic effect, the composition is improved, the suspensibility is better, the self life is longer, no adverse influence on the environment is caused, and the buzurasuppressarianucleopolydro virus and beauveria bassiana insecticidal suspending agent is a safe and nontoxic high efficiency biological pesticide capable of avoiding generation of drug resistance.

The invention relates to a preparation method of a classical swine fever spleen-lymph-sourced compound living vaccine and belongs to the technical field of veterinary biological products. The preparation method of the classical swine fever spleen-lymph-sourced compound living vaccine comprises the following process steps: selecting a domestic rabbit to be inoculated; inoculating; measuring the temperature and observing; harvesting; preparing virus suspension and inspecting; carrying out mixed adsorption on the harvested full-suspension virus bulk and chicken anti-hog cholera virus egg yolk antibody; and adding freezing and drying protecting agent containing an immunopotentiator into antigen antibody compound obtained through adsorption, carrying out split charging, freezing, and carrying out vacuum drying, so that the classical swine fever spleen-lymph-sourced compound living vaccine is obtained. By adopting the preparation method of the classical swine fever spleen-lymph-sourced compound living vaccine, an antibody acquiring route is simple, animal welfare can be improved, manpower and material resources are saved, production cost is greatly reduced, humoral immunity of the living vaccine can be greatly improved, and high neutralizing antibodies can be produced; and the classical swine fever spleen-lymph-sourced compound living vaccine can be applied to piglet immunization, maternal antibody interference can be avoided, no interference is produced to other vaccines, especially a mycoplasma hyopneumoniae vaccine and the like, after the classical swine fever spleen-lymph-sourced compound living vaccine is applied to piglets, and strong mucosal immune response can be induced, so that protection rate is improved.

The invention discloses a construction method of a chronic hepatitis E virus mouse model. In the method, HEV strains are derived from feces of chronically infected rhesus monkeys, and the rhesus monkeys continuously expel toxin for 93 weeks; after an HEV virus suspension is inoculated into BALB/c mice through caudal veins, HEV RNA can be continuously detected in feces and serum of the mice, and the HEV RNA has high virus copying property, and the continuous positive time can reach 36 weeks. The method for establishing the model is simple, can be used for researching the persistent infection mechanism of HEV, and can provide an important animal model for screening and evaluation of anti-HEV drugs.

The invention belongs to the technical field of disinfection, relates to a virus inactivation test, and particularly relates to a removal method of residual disinfectant in the virus inactivation test. The removal method comprises the steps of that (1) a preparation step is performed, specifically, a virus suspension is prepared; (2) a removal step is performed, specifically, first, disinfectant is sucked into a test tube, disinfectant removal treatment is performed after water bath, then ultracentrifugation is performed after 30-50 times dilution, then supernatant is removed, finally a cell maintenance solution is added for dilution and mixing again,, and the residual disinfectant is removed; and (3) a detection step is performed, specifically, virus suspension is sucked and added and mixed well, and a final sample is sucked for virus titer determination. According to the removal method of the residual disinfectant, the test time for disinfectant removal in the whole process can be shortened, the test efficiency of the virus inactivation test can be greatly improved, and the effects of disinfectants and neutralizers on cells and viruses in chemical neutralization methods are avoided.