299results about "Recovery/purification" patented technology

The invention relates to a culture medium for high-density serum-free suspension culture of BHK-21 cells and application of the culture medium. The culture medium contains inorganic salt, amino acid, vitamins, energy substances, a buffer substance, an antioxidant, sterol, polyamine, an additive and an indicator, and the antioxidant comprises dihydrolipoic acid and S-acetyl-L-glutathione. According to the invention, the culture medium for serum-free suspension culture of the BHK-21 cells, which is clear in components and remarkable in culture effect, can be provided. The added dihydrolipoic acid and S-acetyl-L-glutathione are combined as antioxidants of the culture medium, so that the survival rate of BHK-21 cells cultured in the culture medium is improved while the oxidation resistance of the serum-free suspension culture medium is improved.

The present disclosure provides materials and methods related to the purification of viral vectors. In particular, the present disclosure provides peptides, compositions, adsorbents, and related methods, capable of removing process-related impurities (e.g., host cell proteins, nucleic acids, and media components) and product-related impurities (e.g., product fragments, product aggregates, and inactive forms derived from product degradation by or association with other species in the cell culture harvest) from samples during the production and purification of adenovirus.

Provided is a method for large-scale preparation of a purified preparation of a recombinant lentiviral vector at the GMP grade. The method comprises: (a) providing raw material feed liquid to be purified that comprises recombinant viral vectors; (b) carrying out a microfiltration treatment on the feed liquid to obtain a microfiltered filtrate comprising the recombinant viral vectors; (c) optionally concentrating the filtrate to obtain a concentrated filtrate; (d) purifying the filtrate obtained in the previous step by means of chromatography to obtain a crude pure product comprising the recombinant viral vectors; and (e) subjecting the crude pure product obtained in the previous step to liquid exchange and elaborate purification to obtain the purified recombinant viral vectors.

The present disclosure is directed to methods of improving the transduction efficiency of non-transducible cells, such as cells with low hLDLR. The cells may be labeled with 5'FAM and transduced with a scFv-truncated VSV-G pseudotyped lentivirus which comprises an anti-fluorescein scFv. Further provided are methods of gene therapy, cell engineering, genetic screening, and disease models using the fluorescein-labeled cells transduced with the scFv-truncated VSV-G pseudotyped lentivirus.

The invention relates to a preparation method of a Vero cell influenza virus vaccine, and belongs to the technical field of virus culture. In the culture of the influenza virus, red algae protein and soybean protein hydrolysate are added into a culture medium at the same time, the red algae protein and soybean protein hydrolysate are subjected to low-temperature ammonium sulfate precipitation and PD-10 desalination column purification, natural conformation is kept, integrin-FAK-Src and IGF-1R signal channels can be activated, and cell adhesion and proliferation are promoted; the latter contains soybean protein isolate, polysaccharide and RGD motif peptide, and can form an adhesion film on a substrate and mediate adhesion through integrin. According to the invention, the Vero cells and the recombinant plasmid are combined to improve cell adherence, metabolic homeostasis and oxidation resistance, so that the Vero cells grow at high density and efficiently replicate influenza viruses under a serum-free condition, the virus titer (TCID50) reaches 8.5 log10 / mL, and the Vero cells and the recombinant plasmid are suitable for industrial vaccine production.

Methods are disclosed for producing high-titer recombinant adeno-associated virus (rAAV) using a modified adenovirus-based helper nucleic acid. One aspect provided herein is a human adenovirus 5 (hAd)-based nucleic acid that does not contain one or more of (a) an E4 region having E4-ORF6 / 7, (b) a viral associated (VA) RNA region, (c) an E2A region having L4-22K and L4-33K, (d) at least one packaging protein, (e) at least one structural protein, (f) a major late promoter (MLP), (g) an E1 region, and / or (h) an E3 region.

The application discloses a kind of disposable bioreactor and method for preparing adenovirus harvest, belong to biological preparation technical field, the bioreactor includes reaction bag and magnetic stirring mechanism, the magnetic stirring mechanism includes stirring shaft, stirring vane, the number of the stirring vane is greater than 1 and is installed on stirring shaft in layers, the stirring vane includes second stirring vane and first stirring vane, the second stirring vane is fixed on stirring shaft, the first stirring vane is installed on stirring shaft by collar, further include linkage module and shear force transmission device, the linkage module is used to drive shear force transmission device action, realize shear force transmission device and collar connection and disengage.The method is realized based on the bioreactor.Adopting the present scheme, the stirring consistency of disposable bioreactor can be effectively guaranteed in the process of being used for expansion culture.

The application discloses a production method of a high-efficiency strain of Bupalus piniarius nuclear polyhedrosis virus, and belongs to the technical field of biological pesticide production. 8 The virus yield of the high-efficiency strain of Bupalus piniarius nuclear polyhedrosis virus is 6*10 The virus loss rate is reduced by 1.4% by adding 0.1% detergent before centrifugal purification of the virus, and the high yield of the virus is further ensured.The insecticidal effect of the high-efficiency strain produced by the method is increased by more than 64.1% than that of an ordinary strain, and the high-efficiency property is continuously maintained.The method has a very beneficial effect on the production of the high-efficiency strain of Bupalus piniarius nuclear polyhedrosis virus, and provides a solid guarantee for popularization and application of the high-efficiency strain.

A two-step chromatography purification scheme is described which selectively captures and isolates the genome-containing rAAV vector particles from the clarified, concentrated supernatant of a rAAV production cell culture. The process utilizes an affinity capture method performed at a high salt concentration followed by an anion exchange resin method performed at high pH to provide rAAV vector particles which are substantially free of rAAV intermediates.

The application belongs to the technical field of biology, and discloses a kind of Myzus persicae densovirus, amplification and connector method and its application, Myzus persicae densovirus name is Myzus persicae densovirus 2, abbreviated as MpDV2, preserved in China typical culture preservation center, and the preservation number is V202281.The Myzus persicae densovirus MpDV2 provided in the application is a virus strain that has not been isolated and identified before and has pathogenicity to Myzus persicae, and the full-length sequence of the genome of the Myzus persicae densovirus MpDV2 (including the complete terminal inverted repeat sequence) is disclosed for the first time, and the influence of the virus infection on the growth and development of Myzus persicae and the population dynamics / growth.The Myzus persicae densovirus MpDV2 of the application significantly reduces the intrinsic growth rate (r), net reproduction rate (R0) and week-long growth rate (lambda) of Myzus persicae population, and prolongs the average generation time (T) of the population, and the MpDV2 infection has a significant influence on the growth and development of Myzus persicae and population growth.

A method for producing a target protein, comprising: providing a host cell, a vector comprising a nucleic acid sequence encoding the target protein, and a transposase; transfecting the host cell using the vector and the transposase to obtain transfected cells; culturing the transfected cells to form a culture; and harvesting the culture to obtain the target protein from the harvested culture, wherein the cells of the culture are not passaged before the culture is harvested.

The present invention provides: a culture medium composition which contains a pyruvate dehydrogenase kinase inhibitor and is used for promoting the production of a virus having a target gene mounted thereon when culturing a virus-producing cell into which the target gene has been introduced; a culture medium composition for promoting the production of a virus or a virus-like particle when culturing a cell that produces the virus or the virus-like particle; and the like.

A method of purifying a composition comprising a Group B adenovirus, for example comprising the following purification step: diafiltration of a composition comprising the Group B adenovirus using a diafiltration buffer having a conductivity of at least 180 mS cm ‑1 , for example a conductivity of 190, 200, 210, 220, 230, 240, 250, 260, 270, 280 or 290 mS cm ‑1 . Also provided are compositions obtained using the purification methods disclosed herein.

Chromatographic media comprising porous beads having an inner porous core and an outer porous shell are used for the chromatographic separation of enveloped or membrane-bearing biological particles from impurities such as contaminating DNA and / or proteins. The core is capable of binding molecules through hydrophobic interactions; however, the pore size of the shell prevents particles with a size of 20 nm or larger from penetrating into the beads and interacting with the core. Separation is performed at a pH below 7.4. The enveloped or membrane-bearing biological particles may have been subjected to a prior chromatographic capture step. When used for the purification of enveloped viral particles, the method of the present invention has been found to result in a significantly higher rate of infectious viral particles.

Described herein are Zika virus vaccines and compositions and methods of producing and administering said vaccines to subjects in need thereof.

The application discloses a preparation method of a tetravalent influenza virus split vaccine and belongs to the technical field of vaccines. The preparation method of the tetravalent influenza virus split vaccine comprises the following steps: pre-incubation of a chicken embryo, pre-embryo inspection, virus inoculation and culture, post-embryo inspection, cold embryo, virus harvesting, ultrafiltration clarification, zone centrifugation, desugaring, chromatography, virus splitting, virus inactivation, ultrafiltration liquid replacement, sterilization filtration and preparation of a tetravalent influenza virus split vaccine product. Compared with the prior art, the technical means adopted in the application effectively reduces the content of free formaldehyde and improves the antibody level of the vaccine.

The present invention relates to a method for producing a poxvirus viral vector-based vaccine product from a avian cell line. In some embodiments, the avian cell line is a suspended quail cell line. Pharmaceutical compositions, such as vaccines, produced by the method of the present invention are also provided. In some embodiments, the poxvirus viral vector is modified vaccinia virus ankara ("MVA") or recombinant MVA. In some embodiments, the recombinant MVA encodes a heterologous antigen and can be used to produce a vaccine against that antigen. In some embodiments, the recombinant MVA encodes an antigen of respiratory syncytial virus (RSV), and the avian cell line is used to produce a vaccine against RSV containing recombinant MVA and / or the encoded antigen.

The invention discloses a porcine epidemic diarrhea virus G2c genotype strain as well as a separation method and application thereof. Aiming at the current epidemic PEDV variant strain, the genetic variation rule of the epidemic strain is clarified by carrying out virus separation, identification and pathogenicity analysis, and the method has important significance on epidemiology, vaccine development and the like of the PEDV. Specifically, an applicant successfully separates a PEDV strain capable of realizing stable passage propagation in Vero-CCL81 cells from intestinal contents of diarrhea piglets collected from a pig farm of Jiangpermanent Province in Hunan, the PEDV strain is identified and determined as a porcine epidemic diarrhea virus G2c subtype, the PEDV strain is named PEDV CH / HNJY / 2025, the PEDV strain is preserved in China General Microbiological Culture Collection Center on December 11, 2025, and the preservation number of the PEDV strain is CGMCC NO. The preservation number is CGMCC (China General Microbiological Culture Collection Center)

An object of the present invention is to provide a method for improving a proportion of capsids containing a gene of interest GOI in a method for producing a virus, and to provide a method for improving a proportion of capsids containing a gene of interest GOI in a method for introducing a nucleic acid into cells, the method including a nucleic acid introduction step of adding a nucleic acid to a suspension containing cells. According to the present invention, there is provided a method for producing a virus, the method including a nucleic acid introduction step of adding a nucleic acid to a suspension containing cells having a density of 10×106 cells / mL or more, and a culture step of culturing the cells into which the nucleic acid has been introduced, in which the nucleic acid is added in a case where a population cell doubling time of the suspension satisfies 35.4 hours or less.

The present disclosure provides materials and methods related to the purification of viral vectors. In particular, the present disclosure provides compositions, and related methods, comprising peptide ligands capable of removing process-related impurities (e.g., host cell proteins, nucleic acids, and media components) and product-related impurities (e.g., product fragments, product aggregates, and inactive forms derived from product degradation by or association with other species in the cell culture harvest) from biological fluids during the production and purification of adeno-associated viruses (AAVs).

The present application relates to a virus purification method. The present application relates to a method for purification of herpes simplex virus (HSV-1). The present application also relates to HSV-1 purified according to the method. In particular, the method does not comprise an ultrafiltration concentration step. In particular, the method has a high virus titer yield and a high impurity removal rate.

The present invention addresses the problem of providing a virus production method with which it is possible to increase the production of viruses. According to the present invention, provided is a virus production method comprising: a nucleic acid introduction step for introducing a nucleic acid into a cell by electroporation to obtain a cell into which the nucleic acid has been introduced; and a culturing step for culturing the cells into which the nucleic acid has been introduced, the CNET product defined in the specification being 1 * 104 to 1 * 107 (inclusive) in the electroporation.

Providing a method for producing an influenza virus vaccine that does not use eggs. [Solution] Currently, the steps taken before releasing influenza strains to vaccine manufacturers include the step of passage the influenza virus through eggs. The present invention aims to provide a useful procedure in the manufacture of influenza vaccines such that the use of eggs is reduced, and preferably completely avoided. For example, instead of using chicken eggs for influenza vaccine isolation, MDCK cells (Madinderby canine kidney cells) may be used, and the MDCK cells may grow in suspension, in serum-free medium, in protein-free medium, be non-tumoric, grow in the absence of stratified medium, etc.

The present invention relates to methods of producing poxvirus viral vector-based vaccine products from avian cell lines. In some embodiments, the avian cells are suspension quail cell lines. Pharmaceutical compositions such as vaccines produced by the methods of the invention are also provided. In some embodiments, the poxvirus viral vector is Modified Vaccinia Virus Ankara ("MVA") or recombinant MVA. In some embodiments, the recombinant MVA encodes heterologous antigens and can be used to produce a vaccine against the antigens. In some embodiments, the recombinant MVA encodes antigens of Respiratory Syncytial Virus (RSV) and the avian cells are used to produce a vaccine against RSV comprising the recombinant MVA and / or the encoded antigens.

The application provides a bacteriophage for controlling Vibrio infection, and belongs to the field of veterinary technology. The application designs a recombinant tail chain protein rTC and synthesizes an expression gene of the recombinant tail chain protein rTC through amino acid sequence alignment and structure analysis of tail chain proteins of Vibrio phages. The rTC gene is electroporated into Vibrio parahaemolyticus isolated from diseased prawns, and the host bacteria are infected with PHxp1 bacteriophage isolated from seawater; after the rTC gene is recombined with the PHxp1 bacteriophage, the collected bacteriophage is screened with Vibrio alginolyticus to obtain two bacteriophages PHxp1-1 and PHxp1-2 capable of lysing Vibrio alginolyticus; the bacteriophage is screened with Vibrio harveyi to obtain one bacteriophage PHxp1-3 capable of lysing Vibrio harveyi. The host spectrum of the three recombinant bacteriophages is widened. The recombinant tail chain protein bacteriophage greatly reduces the death and growth stagnation of prawns caused by Vibrio infection in prawn farms, and improves the breeding benefit.

The present invention relates to a phage-based gene delivery system targeting human cells or animal cells, including immune cells, other human cells and cancer cells. In particular, the present invention relates to novel phage particles and associated phagemid expression systems and their production for the delivery of transgenes, comprising DNA encoding pathogen antigens, to human immune cells, suitable for use in vaccines and vaccine development against infectious diseases and cancers.

A method for large-scale preparation of a purified GMP-grade recombinant lentiviral vector preparation is provided, comprising the following steps: (a) providing a raw material, a feed solution having a volume Va, containing the recombinant viral vector to be purified; (b) subjecting the feed solution to a microfiltration process to obtain a microfiltrate having a volume Vb, containing the recombinant viral vector; (c) optionally concentrating the filtrate to obtain a concentrated filtrate having a volume Vc; (d) purifying the filtrate obtained in the previous step by chromatography to obtain a crude product containing the recombinant viral vector; and (e) liquid-exchanging and finely purifying the crude product obtained in the previous step to obtain a purified recombinant viral vector.

The invention discloses a method for rapidly separating and screening soil bacteriophages, and belongs to the field of microbial technology. The method comprises the following steps: mixing a soil sample with an SM buffer solution, adding a CaCl2 solution and an LB solution, and carrying out enrichment culture on a plurality of logarithmic phase host bacteria; centrifuging, taking supernatant and filtering to obtain phage suspension; primarily screening plaque through a spot method or a double-layer plate method, and carrying out multiple rounds of gradient dilution and purification to obtain a single phage; eluting the amplified plaque by using an SM buffer solution, and filtering to obtain a phage stock solution; adding a ferric chloride solution to the stock solution to precipitate bacteriophages, centrifuging, and oscillating and releasing with an ascorbic acid mixed solution; dNase I and a buffer solution are added to digest free nucleic acid, and after the reaction is terminated, a soil DNA extraction kit is utilized to obtain bacteriophage genome DNA.

This disclosure provides a modified rep-cap plasmid containing a tetracycline-inducible expression system that can be used to produce a recombinant AAV1 vector.

The invention relates to a method for purifying retrovirus, and particularly, the method comprises affinity chromatography, composite mode chromatography and molecular sieve chromatography. The retrovirus purified by the method is high in titer recovery rate, excellent in impurity removal effect, low in requirement on an initial to-be-purified sample and simple in amplification of a purification process, and can be used for large-scale purification of the retrovirus, and the yield and the purity of the retrovirus meet industrial requirements.