67 results about "Vero cell" patented technology

The invention relates to a method for inhibiting in-vitro infection of a porcine epidemic diarrhea virus by using JPH203 and application. The JPH203 can be used for preparing an antiviral drug for inhibiting infection of the porcine epidemic diarrhea virus. JPH203 with the concentration of 5 [mu] M is added in the process that Vero cells are infected with PEDV, compared with a control group without JPH203, the virus infection condition, the protein expression quantity, the RNA content and the virus titer of the PEDV in a treatment group with JPH203 are all remarkably reduced, and it is indicated that JPH203 can be used for inhibiting in-vitro infection of the porcine epidemic diarrhea virus.

The invention relates to a Vero cell serum-free culture medium and application thereof, the serum-free culture medium is suitable for free suspension growth of Vero cells and is free of animal sources and protein components, adherent cells can be directly domesticated through a certain method to screen out Vero suspension cells, and serum-free suspension culture of the Vero suspension cells is achieved in the serum-free culture medium. The density of cells subjected to suspension culture by adopting the serum-free culture medium can reach more than 107 cells / mL, the cell viability is more than 90%, the cells are well dispersed, the cells can still normally grow and proliferate after continuous subculture for 20 generations, and the cells are sensitive to various viruses.

The invention belongs to the technical field of gene detection of cross fusion of biology and a new generation of information technology, and particularly relates to a method for detecting residual DNA of multiple host cells based on digital PCR (Polymerase Chain Reaction). The invention discloses a multiplex digital PCR quantitative detection primer pair and a probe group for detecting residual DNA of CHO cells, Vero cells, pichia pastoris, escherichia coli and NS0 cells for the first time. By utilizing the method, not only can the residual DNA of five host cells be simultaneously subjected to single or multiple detection in a single reaction, but also non-specific amplification, signal interference and detection limitation of the residual DNA of the host cells in the qPCR detection process can be solved friendly, and meanwhile, the original concentration of a sample is absolutely quantified by the method; the method does not depend on the establishment of a standard curve, reduces the influence caused by a standard product, and is superior to the prior art.

The invention discloses an application of Arcyriaflavin A or a pharmaceutically acceptable salt thereof in preparation of a medicine for preventing and treating monkey pox virus infection. The invention finds that the Arcyriaflavin A can obviously inhibit the replication of the monkey pox virus in a Vero cell, and the IC50 of the Arcyriaflavin A for inhibiting the Ib branch monkey pox virus and the IC50 of the Arcyriaflavin A for inhibiting the IIb branch monkey pox virus are 0.66 mu M and 0.96 mu M respectively. The invention also verifies that the Arcyriaflavin A has extremely low toxicity to Vero cells, and the toxicity CC50 of the Arcyriaflavin A to the cells is 41.51 mu M when the Arcyriaflavin A is used for inhibiting the proliferation of the monkey pox virus, so that the Arcyriaflavin A can be used for effectively inhibiting the replication of the monkey pox virus within the safe use range of the Arcyriaflavin A. The invention provides a new choice for the prevention and treatment of monkey pox, and has important significance for the prevention and treatment of monkey pox.

The invention discloses an akabane virus inactivated vaccine preparation method, which comprises: carrying out Vero cell passage in a cell bottle, taking well growing cells until the cells grow to 80-90%, adding into a DMEM culture medium containing 10% of fetal calf serum, taking out the cells growing to 80-90% and having a good state, adding akabane virus CH-JL-01-2022 according to a volume ratio of 0.1%, and carrying out culture at a temperature of 80-90 DEG C so as to obtain the akabane virus inactivated vaccine. Culturing for 2-3 days in an incubator with the temperature of 37 DEG C and 5% CO2; the method comprises the following steps: preparing 0.25 M BEI, 0.1 M BPL and a 40% formaldehyde solution as inactivators, setting five concentration gradients according to the volume ratios of 0.01%, 0.05%, 0.1% and 0.5%, and collecting after 6 hours, 12 hours, 24 hours, 36 hours and 48 hours after adding; the screening of the akabane virus inactivation conditions according to the types, concentrations and inactivation time of inactivators finds that 0.5% formaldehyde shows a complete inactivation effect after inactivation for 36 hours. The inactivated virus liquid is mixed with MONTANIDETM ISA 201, an ImjectI aluminum adjuvant and a manganese adjuvant according to the volume ratio of 1: 1, and then immunization is carried out, and the protection effect of the ImjectI aluminum adjuvant is found to be optimal.

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 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.

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 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 present invention relates to specific monoclonal antibodies and their applications. The amino acid sequence of the light chain CDR1 of the monoclonal antibody is shown in SEQ ID No. 1, the amino acid sequence of CDR2 is shown in SEQ ID No. 2, and the amino acid sequence of CDR3 is shown in SEQ ID No. 3; the amino acid sequence of the heavy chain CDR1 of the monoclonal antibody is shown in SEQ ID No. 4, the amino acid sequence of CDR2 is shown in SEQ ID No. 5, and the amino acid sequence of CDR3 is shown in SEQ ID No. 6. The present invention uses recombinant monkeypox virus surface protein E8L as an antigen. The screened monoclonal antibodies have good affinity for the recombinant monkeypox virus surface protein E8L at the molecular and cellular levels, can significantly inhibit the infection efficiency of VACV replication-deficient strains in VERO cells, and have good in vitro neutralization effect against monkeypox 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.

The invention provides an application of Akti-1 / 2 in preparation of an anti-new coronavirus drug. And the new coronavirus is SARS-CoV-2 (Sarcabose Region The chemical name of the Akti-1 / 2 is 1, 3-dihydro-1-[1-[4-(6-phenyl-1H-imidazo [4, 5-g] quinoxaline-7-yl) phenylmethyl]-4-piperidinyl]-2H-benzimidazole-2-ketone, in-vitro Vero cell infection model research shows that the Akti-1 / 2 can inhibit replication of an original strain of the new coronavirus (SARS-CoV-2), and the activity of resisting the new coronavirus is achieved. In a Vero cell infection model, the IC50 (half maximal inhibitory concentration) of the SARS-CoV-2 original strain is 0.039 [mu] M. Therefore, the Akti-1 / 2 can be used for research and development of anti-new coronavirus infection drugs, and has a good development prospect.

The invention relates to the technical field of biology, in particular to Chinese yam active peptide and a preparation method and application thereof.The Chinese yam active peptide comprises active peptide and free amino acid, the mass content of the active peptide with the molecular weight of 180-1000 Da is larger than or equal to 65%, and the mass content of hydrophobic amino acid in the free amino acid is larger than or equal to 30%. The preparation method of the Chinese yam active peptide is easy and convenient to operate, through the synergistic effect of the neutral protease and the alkaline protease, on the premise that the hydrolysis degree of the Chinese yam peptide is improved, it can be prevented that a large number of amino acids are generated through excessive hydrolysis, glycoprotein in Chinese yam is subjected to sufficient enzymolysis, and the yield of the Chinese yam active peptide is increased. The Chinese yam peptide prepared by the method can protect the vitality of Vero cells and MRC5 cells, has the characteristics of high activity, small molecular weight and easiness in absorption, and can be widely applied to the fields of medicines, foods, health care products, cell culture additives and the like.

The invention discloses application of iso-pasteuretin in preparation of a medicine for treating diseases caused by methicillin-resistant staphylococcus aureus, and belongs to the technical field of biological medicine. According to the present invention, it is identified that the snowfield tea endophytic fungus 202109-Ts-F016 is the new Coprinopsis thamnolicola under coprinus pseudoratus, three monomeric compounds with anti-MRSA activity are separated from the secondary metabolite of the Coprinopsis thamnolicola, the three compounds can be adopted as the candidate inhibitors of the novel antibacterial target FtsZ, and the toxicity of the pasteurestin C and the iso-pasteurestin on the Vero cell is low. The invention provides an important basis and a material basis for the development of coprinus fungus metabolites and anti-MRSA new target drugs.

The present invention discloses a rabies virus serum-free Vero cell inactivated vaccine and a preparation method thereof, and belongs to the technical field of vaccine manufacturing. The preparation method of the inactivated vaccine comprises the following steps: resuscitating Vero cells, performing serum-free adaptation and domestication of the Vero cells, preparing modified microcarriers, adding the modified microcarriers to a bioreactor, adding serum-free culture medium, inoculating the domesticated Vero cells into the bioreactor, amplifying and harvesting the cells, inoculating rabies virus, adding serum-free culture medium for virus culture, harvesting, and obtaining a virus liquid; finally, inactivating the virus, incubating, concentrating, purifying, and freeze-drying to obtain a rabies virus serum-free Vero cell inactivated vaccine. The virus liquid prepared by the present invention has the advantages of high virus content, stable production process, and low production cost, and the prepared vaccine has the effects of improving safety in use and reducing side effects.

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 invention belongs to the field of interferon genetic engineering, and discloses recombinant pigeon alpha interferon and its application in the preparation of medicines for resisting pigeon paramyxovirus infection. The present invention constructs recombinant pigeon interferon piINF 5, realizes efficient expression and purification in CHO cell expression system, significantly prolongs its half-life and improves stability. Recombinant pigeon interferon piINF 5 has no toxic effect on DF1 and VERO cells, and can effectively stimulate the transcription level of interferon-stimulated genes, can significantly inhibit the replication of pigeon paramyxovirus, reduce viral copy number and hemagglutination value. In vivo animal attack experiment shows that the recombinant pigeon interferon piINF 5 can well protect pigeons from the attack of pigeon paramyxovirus, and the survival rate reaches 80%, and there is no detoxification in the first five days after the attack, and only 20% of animals have detoxification in the later stage. The present invention provides an efficient and stable new drug selection for the treatment of pigeon paramyxovirus disease, with good commercial application prospects.

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 discloses a human-sheep rotavirus gene reassortment attenuated strain LG1 and application of the human-sheep rotavirus gene reassortment attenuated strain LG1. The human-sheep rotavirus gene reassortment attenuated strain LG1 comprises: i) a coding gene of a structural protein VP7 of a human G1 type rotavirus HN15D2 strain; and ii) coding genes of structural proteins VP1, VP2, VP3, VP4 and VP6 and non-structural proteins NSP1, NSP2, NSP3, NSP4 and NSP5 of the sheep rotavirus strain LLR. The human-sheep rotavirus gene reassortment strain which takes a sheep rotavirus strain as a skeleton, contains a human G1-VP7 gene and can be stably passaged on a Vero cell is successfully constructed for the first time, and the reassortment strain not only has the safety of an animal strain, but also has the immunogenicity of a human G1 type RV, can effectively prevent infection of a human G1 type wild strain, and has a good application prospect. The method has important significance for developing multivalent RV vaccines containing G1 genotypes and developing RV vaccines which are higher in prevention pertinence and provide a better protection effect on Chinese infantile diarrhea.

The invention discloses a serum-free culture medium for culturing Vero cells. The serum-free culture medium comprises a DMEM (Dulbecco Modified Eagle Medium), an ultrafiltration type animal-source-free soybean protein hydrolysate and a TE02 additive, the content of the DMEM culture medium is 13380 mg / L, the content of the ultrafiltration type animal-source-free soybean protein hydrolysate is 1000 to 5000 mg / L, and the content of the TE02 additive is 0.2 to 0.5 mL / L; the serum-free culture medium disclosed by the invention is superior to commercial like products in cell growth morphology, amplification multiple and passage stability; the serum-free culture can be completely supported in the cell culture process, so that the influence caused by the quality difference between different batches of serum is avoided, and the blood-derived pollution is avoided; the batch-to-batch difference in the large-scale production process is easy to control, the culture medium is stable in property, and the transportation and preservation conditions are relatively loose; the feed is clear in components, free of animal-derived components, low in protein content and high in safety.

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.