39 results about "Vero cell" patented technology

The invention provides a Garlu virus attenuated strain and application thereof, and relates to the field of virology and biological medicine. According to the invention, the screened Garlu virus attenuated strain is a natural attenuated strain derived from a tick sample, can grow in Vero cells, has the titer of more than or equal to 107 TCID50 / mL, does not have clinical pathogenicity to an immunodeficient host, and is safe; after the attenuated strain is used as an immunogen for pre-intervention, infection of a lethal virus strain can be avoided, the survival rate is increased to 100%, adverse effects on growth and life of experimental animals are avoided, particularly, a host can be stimulated to generate humoral immunity and cellular immunity at the same time, and the survival rate of the experimental animals is improved. And moreover, broad-spectrum cross-protection efficacy can be generated on Bandavirus lethal virus strains, and a foundation is laid for improvement of research on Bandavirus live vaccines such as the Colu virus and the like.

The invention provides an influenza virus that demonstrates enhanced growth in Vero cells. The influenza virus includes PA, NP, and NS gene segments having selected nucleotides and encoding proteins having amino acid sequences with selected amino acids. The invention also provides a pharmaceutical formulation containing the influenza virus, as well as a method of eliciting an immune response in a mammal by administering the influenza virus to the mammal, and a method for generating the influenza virus.

The invention provides an application of CC-115 in preparation of an anti-new coronavirus drug. The chemical name of the CC-115 is 1-ethyl-7-[2-methyl-6-(1H-1, 2, 4-triazole-3-yl) pyridine-3-yl]-3, 5-dihydropyrazino [2, 3-b] pyrazine-2 (1H)-ketone, and the structural formula of the CC-115 is shown in the description. An in-vitro Vero cell infection model finds that the CC-115 can inhibit the replication of a new coronavirus (SARS-CoV-2) original strain and an Omicro variant (EG.5.1), and the IC50 (half maximal inhibitory concentration) of the CC-115 aiming at the SARS-CoV-2 original strain is 0.004 [mu] M and 0.005 [mu] M. The CC-115 has the advantages that the CC-115 can inhibit the replication of the SARS-CoV-2 original strain and the Omicro variant (EG.5.1); the CC-115 can be used for research and development of anti-new coronavirus infection drugs, and has a good development prospect.

The present invention relates to a low-serum medium composition for culturing Vero cells, and a method for culturing Vero cells and a method for producing a virus, both using the same.

The invention belongs to the technical field of traditional Chinese medicine (Tibetan medicine), and particularly relates to a Tibetan medicine composition for treating new coronavirus infection and application of the Tibetan medicine composition. The invention discloses a Tibetan medicine composition for treating new coronavirus infection and application of the Tibetan medicine composition. The composition is prepared from the following components in parts by weight: 100 to 700 parts of fructus chebulae, 10 to 80 parts of radix aconiti lateralis, 10 to 60 parts of radix aucklandiae, 50 to 600 parts of rhizoma acori calami, 10 to 50 parts of Muslim myrrh, 0.01 to 50 parts of calculus bovis factitius and 0.01 to 50 parts of artificial musk. The composition can inhibit the mRNA expression of the N gene and the ORF1ab gene after the SARS-CoV-2 virus infects the Vero cell, so that the treatment on the novel coronavirus infection is realized.

This invention discloses a strain of Rhodotorula glutinis I7Y2 that promotes cell proliferation in a serum-free environment and its applications. Rhodotorula glutinis ( Rhodotorula mucilaginosa The yeast strain I7Y2 was deposited at the Guangdong Provincial Microbial Culture Collection Center (GDMCC) on March 13, 2026, with accession number GDMCC No: 67951. This invention has a good effect on promoting Vero cell proliferation, specifically manifested in: (1) the yeast involved in this invention can produce various active substances such as amino acids, polypeptides, sanshool, and inositol to meet the nutritional needs of cell growth; (2) it can effectively improve the viability and density of Vero cells in the basal culture medium; (3) it can effectively improve the proliferation rate and migration rate of Vero cells in the basal culture medium; and (4) it can effectively reduce the apoptosis rate of Vero cells and the expression of apoptosis-related genes in the basal culture medium. Therefore, Rhodotorula glutinis I7Y2 has great application potential in promoting cell proliferation, especially in serum-free culture products.

The invention discloses an influenza A virus Vero cell cold adapted strain Vca-CPD and application thereof, and relates to the field of biotechnology and vaccinology. The preservation number of the Vca-CPD provided by the invention is CCTCC (China Center For Type Culture Collection) NO: V202585. The strain is mainly obtained by performing codon pair optimization on PB2, PA and NP genes. In-vivo and in-vitro experiments prove that the replication ability of the strain in human cells and mammal models is obviously weakened, and the safety is improved; meanwhile, an organism can be induced to generate high-level and specific serum protective antibodies and respiratory mucosa sIgA antibodies. In addition, the virus yield of the strain in production of stromal cells is equivalent to that of a parent strain, and the strain shows good production applicability.

The invention discloses a primer probe combination for detecting porcine parvovirus type 8 and application of the primer probe combination, and relates to the technical field of porcine parvovirus detection. The primer probe combination comprises a forward primer, a reverse primer and a detection probe, the nucleotide sequence of the forward primer is as shown in SEQ ID NO.1, the nucleotide sequence of the reverse primer is as shown in SEQ ID NO.2, and the nucleotide sequence of the detection probe is as shown in SEQ ID NO.3. The primer probe combination disclosed by the invention is high in specificity when being used for detecting PPV8, and has no cross reaction with a non-PPV8 genome; the sensitivity is high, and the detection limit of a PPV8 plasmid standard substance is 100 copies / reaction (5copies / microliter); the durability and matrix interference resistance are high, and detection can also be performed after a 293T / CHO / Vero cell genome is introduced.

The invention relates to a Rosa26 site-targeted Vero cell line as well as a construction method and application thereof, and belongs to the technical field of gene engineering. The invention provides a Vero cell line targeting a Rosa26 site. The Vero cell line comprises an attP sequence integrated into the Rosa26 site of a genome of the Vero cell line. Researches show that by co-transfecting donor plasmids expressing exogenous genes and attB sites and Bxb1 expression plasmids to the Vero cell line, the exogenous genes can be rapidly integrated to a genome of the Vero cell line, and then the Vero stably transfected cell line capable of stably expressing the exogenous genes for a long time is constructed. According to the Vero cell line provided by the invention, the construction time of the Vero stable transfection cell line for expressing an exogenous gene is greatly shortened, and based on insertion of the Rosa26 site, the Vero cell line provided by the invention can maintain the stability and consistency of a genome for a long time, so that long-term stable expression of the exogenous gene is ensured.

The present application relates to a kind of preparation methods of Vero cell influenza virus vaccine, belong to the technical field of virus culture.In the culture of influenza virus, medium is added with red algae protein and soybean protein hydrolysate, the former is precipitated by low-temperature ammonium sulfate and purified by PD-10 desalting column, maintains native conformation, can activate integrin-FAK-Src and IGF-1R signal pathway, promotes cell adhesion and proliferation;The latter contains soybean protein isolate, polysaccharide and RGD motif peptide, can form adhesive film on the substrate and adhere through integrin mediation.Both of them improve cell adhesion, metabolic homeostasis and antioxidant capacity, so that Vero cell can grow in high density under serum-free condition and efficiently replicate influenza virus, virus titer (TCID 50 ) reaches 8.5 log 10 / mL, suitable for industrialized vaccine production.

The invention provides a low-autophagy Vero cell line based on a DDIT3 inhibition gene as well as a construction method and application of the low-autophagy Vero cell line. The cell line stably knocks down DDIT3 expression through lentivirus-mediated shRNA, the autophagy level under the condition of nutrition deficiency is remarkably reduced, the cell proliferation capacity is enhanced, and the yield of influenza A H1N1 viruses is remarkably increased. The construction method comprises the following steps: cloning shRNA (short hairpin ribonucleic acid) to a GV493 lentiviral vector, co-transfecting 293T cell packaging virus with psPAX2 / pMD2. G helper plasmid, infecting Vero cells, and screening through puromycin to obtain a stable cell line. The cell line solves the problem of growth stagnation caused by over-activation of autophagy in serum-free suspension culture of traditional Vero cells, and is suitable for large-scale vaccine production.

The invention discloses application of isolorydine hydrochloride in preparation of antiviral drugs, and belongs to the technical field of biological medicines. The invention discloses the antiviral effect, especially the PEDV (porcine epidemic diarrhea virus) resisting effect of the isolorydine hydrochloride for the first time. On Vero cells infected by PEDV (porcine epidemic diarrhea virus), 100 [mu] M of isolorydine hydrochloride has no cytotoxicity, and the median inhibitory concentration IC50 of the isolorydine hydrochloride is 6.78 [mu] M. In a host cell IPEC-LD, the isolorydine hydrochloride has no cytotoxicity when the isolorydine hydrochloride is 100 [mu] M and shows slight toxicity when the isolorydine hydrochloride is 200 [mu] M, 5 [mu] M of isolorydine hydrochloride can significantly reduce the mRNA level of PEDV N, and 10 [mu] M of isolorydine hydrochloride can significantly inhibit expression of PEDV N protein. The compound is used for preparing the medicine for resisting the porcine epidemic diarrhea virus, and has the advantages of reliable curative effect, no toxic or side effect, abundant resources, low price and the like.

The invention relates to the field of antiviral drugs, in particular to application of a miRNA (micro Ribonucleic Acid) simulant in preparation of anti-I-type herpes simplex virus drugs. The miRNA simulant provided by the invention is an analogue of I-type herpes simplex virus coding miR-H5-5p and is double-stranded RNA synthesized by a chemical method, the molecular weight of the miRNA simulant is about 13300 Daltons, and the sequence of the miRNA simulant is shown as SEQ ID NO. 1 or SEQ ID NO. 2. The miRNA simulant disclosed by the invention can show remarkable activity of resisting the acyclovir-resistant I-type herpes simplex virus in HEK293T cells and Vero cells, so that the miRNA simulant can be used for preparing the medicine for resisting the acyclovir-resistant I-type herpes simplex virus.

The invention relates to the fields of genetic engineering and molecular biology, in particular to a method for optimizing a nucleotide sequence of an expression protein in a host cell and a method for optimizing a nucleotide sequence of an expression protein in a Vero cell. The invention also relates to nucleic acid molecules, vectors, host cells and viroid particles comprising the optimized nucleotide sequences obtained by said method.

The invention discloses an upstream process for producing a virus. The process includes cell expansion of adherent cell cultures grown on cell culture containers and on microcarriers in bioreactors. In particular, a scalable process for the upstream manufacture of a virus is described, which includes optimization of the amount of cell detachment agent used per surface area of a microcarrier to enhance cell growth in production bioreactors.

The invention relates to neurobiology, molecular virology, biomedical engineering and the like, and discloses a domesticated low-serum Vero cell line and a method for preparing an HSV virus vector through suspension culture cell amplification. Aiming at the bottleneck problem that a process for amplifying HSV by adherent culture of Vero cells is difficult to realize large-scale and engineering amplified production, the invention researches and develops a method for preparing HSV based on suspension culture of cell amplification. Vero cells introduced from ATCC are domesticated, and the serum content of a culture medium is gradually reduced, so that the Vero cells are suitable for suspension culture in a microcarrier or sheet carrier reactor by using a low-serum or serum-free culture medium. The method comprises the following steps: using Tamp of Dibide Bioengineering (Shanghai) Co., Ltd; and carrying out suspension culture of the Vero cells and infection amplification of the HSV virus vector by using a J Cloud Ready TM bioreactor. High-activity suspension culture and HSV amplification of Vero cells are realized by optimizing production conditions, cell density, biochemical detection and the like, and a stable and efficient method is provided for preparing a high-titer and high-purity HSV virus vector in combination with purification process upgrading.

The invention relates to a Vero cell line capable of stably expressing TMPRSS2 as well as a construction method and application of the Vero cell line, and belongs to the technical field of biology. The invention provides a Vero cell line capable of stably expressing TMPRSS2. The cell line contains a TMPRSS2 coding gene which is integrated into a Rosa26 site of a cell genome of the cell line. The construction method of the cell line comprises the following steps: firstly, connecting an open reading frame cDNA sequence for coding TMPRSS2 with a donor vector carrying an attB site, then co-transfecting the donor vector and a Bxb1 expression vector into a Vero cell line, and carrying out antibiotic pressurized screening to obtain the stably expressed cell line. The cell line provided by the invention provides reliable candidate stromal cells for influenza virus cells independent of pancreatin culture, and has good application value for separation of influenza viruses and development of novel influenza vaccines.

The invention provides a pyridone-containing aminothiazole heterocyclic compound as well as synthesis and application thereof. The aminothiazole heterocyclic compound has a structure as shown in a general formula I or a general formula II or a pharmaceutically acceptable salt or an isotope labeled compound thereof. The invention also discloses a preparation method and the like of the derivative. Experiments prove that the aminothiazole heterocyclic compound containing pyridone can effectively inhibit replication of herpes simplex virus in Vero cells, the compound is of a non-nucleoside structure, the inhibitory activity of the compound is superior to that of a clinical first-line drug acyclovir, and the compound can be used for preparing active drugs for resisting the herpes simplex virus. The general formula I or the general formula II is shown in the specification.

The invention relates to animal virology and genetic engineering technologies, and aims to provide a recombinant porcine epidemic diarrhea virus with enhanced replication capability. The virus is obtained by modifying a porcine epidemic diarrhea virus PEDV, and the amino acid sequence of nucleocapsid protein (N protein) of the virus is shown as SEQ ID NO: 2. The strain of the virus is preserved in the China Center for Type Culture Collection, the preservation name is rcPEDV P120-NT139A, and the preservation number is CCTCC NO: V202562. According to the invention, threonine at the 139th site of nucleocapsid protein of PEDV is a key modification site influencing PEDV infection, and the new strain can be successfully infected in PEDV infection model verification experiments of IPEC-J2 and Vero cells; the strain shows stronger replication ability in a PEDV infection model verification experiment of IPEC-J2 cells, the production difficulty is reduced, and the strain can be further used for preparing a porcine epidemic diarrhea virus inactivated vaccine.

The present invention relates to a sVERO 7C2 cell line, which is a Vero cell line derived from WHO distributed Vero cells (African green monkey kidney cell line) and capable of being cultured in suspension without serum components. In addition, the present invention relates to a culture method for culturing Vero cells and a method for preparing a virus for vaccine using the Vero cells.

This invention belongs to the field of genetic engineering technology and discloses a shRNA that targets and knocks down the HDAC5 gene. The sense strand of the shRNA is shown in SEQ ID NO.4, and the antisense strand is shown in SEQ ID NO.5. The sense and antisense strands anneal to form a double-stranded DNA with sticky ends. This invention utilizes lentivirus-mediated RNA interference technology to obtain a Vero cell line with stable HDAC5 gene knockdown, demonstrating that HDAC5 gene knockdown can inhibit PPRV proliferation, providing a novel potential target for the development of anti-PPRV drugs.

The invention relates to the technical field of biology, in particular to application of a G3BP1 gene as a target spot in screening of drugs for inhibiting IBV (infectious bronchitis virus) replication, experimental results show that when G3BP1 in cells is deleted, the IBV replication ability is enhanced, it is prompted that G3BP1 possibly has an inhibiting effect on IBV replication, the expression level of G3BP1 is remarkably and negatively correlated with the IBV replication ability, and the G3BP1 gene can be used as a target spot in screening of drugs for inhibiting IBV replication. EGCG can effectively inhibit duplication of IBV in Vero cells by up-regulating expression of a host limiting factor G3BP1, and after infection, administration and preventive administration both show a remarkable antiviral effect. The EGCG has the best excitation effect on G3BP1 and the most obvious inhibition effect on IBV intracellular replication, and the EGCG can effectively reverse IBV infection induced stress particle formation inhibition caused by G3BP1 knock-down.

A preparation method of an application of a sCAR1 mutant nanoparticle against CV-B3 infection are provided, thereby solving a problem of the current lack of preventive vaccines and specific therapeutic drugs for the CV-B3 infection. A sCAR1 Mu+Ferritin nanoparticle that fully exhibits sCAR1 Mu is constructed, and IC50 values of the sCAR1 Mu+Ferritin nanoparticle against the CV-B3 infection in Vero cells and SH-SY5Y cells are 413.05 ng / mL and 140.93 ng / mL, respectively. When 200 μg of sCAR1 Mu+Ferritin is injected intraperitoneally 6 hours prior to an intraperitoneal injection of a lethal dose of CV-B3, it significantly reduces viral loads in liver, spleen, brain, and heart tissues, effectively alleviates inflammatory pathology in heart and brain tissues, and provides a 68.75% survival rate.

The invention relates to the technical field of biology, in particular to application of epigallocatechin gallate in preparation of a medicine for inhibiting infectious bronchitis virus (IBV) replication, and experimental results show that EGCG can effectively inhibit IBV replication in Vero cells by up-regulating expression of a host limiting factor G3BP1, so that the effect of inhibiting IBV replication is achieved. And after infection, administration and preventive administration both show an obvious antiviral effect. The EGCG has the best excitation effect on G3BP1 and the most obvious inhibition effect on IBV intracellular replication, and the EGCG can effectively reverse IBV infection induced stress particle formation inhibition caused by G3BP1 knock-down.

The invention provides an antibody-exosome, a preparation method thereof and application of the antibody-exosome in preparation of ricin detoxification drugs, and belongs to the technical field of drug delivery and intracellular therapy. The antibody-exosome disclosed by the invention comprises a milk-derived exosome (mExo) and a single-domain antibody V9E1 of an anti-ricin A chain; the amino acid sequence of the V9E1 is as shown in SEQ ID NO. 1. V9E1 is loaded into a milk exosome by using a saponin perforation method to obtain a V9E1 antibody-exosome, and the antibody-exosome shows reliable biological safety in both cells and animal bodies; the cell survival rate can be obviously improved in a Vero cell acute poisoning model; after mice are poisoned by ricin for 2 hours and 6 hours, the survival rate of the mice can be remarkably improved; through continuous administration, all mice injected with ricin with a complete lethal dose can survive, liver and spleen tissues can be protected, and the method has the potential of replacing a traditional ricin antibody therapy.

This invention discloses a culture medium and method suitable for large-scale full-suspension culture of Vero cells. The culture medium is a serum-free medium specifically designed and developed based on the growth and metabolic characteristics of Vero suspension cells and their different proliferation abilities when cultured in a reactor versus a shaker, combined with the differences in culture scale. It includes the following components: amino acids, proteins, vitamins, inorganic salts, serum replacement factors, and auxiliary components. Furthermore, this invention determines the control parameters of the full-suspension culture reactor based on cell growth characteristics, significantly increasing the VERO cell culture density. The cell doubling rate reaches 10-fold after 48 hours of culture, the cell production cycle is halved, and production efficiency is improved. The operation is simple, reducing the contamination rate in large-scale culture and solving the problem of long perfusion culture cycles. This is of great significance for the large-scale production of viral vaccines using VERO cells as a substrate.

The invention discloses a monoclonal antibody specifically combined with monkey pox B6R protein and application of the monoclonal antibody, and belongs to the technical field of biology. In order to solve the problem that an antibody with a good effect on monkey pox virus is lacked in the prior art, the invention provides a monoclonal antibody specifically combined with monkey pox B6R protein, and the monoclonal antibody comprises a light chain variable region, a heavy chain variable region, and a heavy chain constant region and / or a light chain constant region. The monoclonal antibody provided by the invention has good affinity with a recombinant monkey pox virus surface protein B6R, can inhibit the infection efficiency of VACV replication defective strains and MPXV in VERO cells, and has a good monkey pox virus in-vitro neutralization effect.

The application relates to the technical field of animal infectious disease prevention and control, and particularly discloses application of tetrandrine in preparation of a medicine for preventing and treating pseudorabies of pigs. The application can significantly inhibit the replication efficiency of PRV on Vero cells and IPAM cells. When the concentration of the inhibitor is higher than 4 muM, the progeny virus titer and the virus gene transcription level of PRV are significantly reduced compared with those of a control group, and the pathological changes caused by cell infection are effectively relieved, which indicates that the tetrandrine can effectively inhibit the proliferation of PRV and has a potential application prospect.

The invention belongs to the technical field of biological medicine. The invention relates to a Vero cell vaccine quality monitoring method, in particular to a Vero cell vaccine quality monitoring method based on mass spectrometry and characteristic host cell protein combination. The invention provides a protein combination and provides a Vero cell vaccine quality monitoring method based on the protein combination, the ratio K of the total abundance of the protein combination to the total abundance of host cell protein (Vero cell protein) is used as a core quality index, the limitation that traditional ELISA only measures the total amount and depends on an antibody is broken through, and high-sensitivity recognition and early warning of HCP spectrum composition deviation are achieved; the method is based on LC-MS / MS non-targeted quantification, does not need antibody detection, avoids antibody cross reaction interference, is high in sensitivity, is suitable for batch-to-batch consistency evaluation, process change evaluation and quality analysis of Vero cell matrix products such as rabies vaccines, provides a new standardized path for whole life cycle quality control of biological products, and has a wide application prospect. Good application prospects and application values are realized.

The invention provides an influenza virus that demonstrates enhanced growth in Vero cells. The influenza virus includes PB1, PB2, PA, NP, and NS gene segments encoding proteins having amino acid sequences with selected amino acids. Optionally, at least one of the PB1, PB2, and PA gene segments includes a cytosine to uracil promoter mutation at nucleotide position 4. The invention also provides a pharmaceutical formulation containing the influenza virus, as well as a method of eliciting an immune response in a mammal by administering the influenza virus to the mammal, and a method for generating the influenza virus.