133results about How to "Increase infectivity" patented technology

The present invention relates to the production of adeno-associated viral vectors in insect cells. The insect cells therefore comprise a first nucleotide sequence encoding the adeno-associated virus (AAV) capsid proteins, whereby the initiation codon for translation of the AAV VP1 capsid protein is a non-ATG, suboptimal initiation codon. The insect cell further comprises a second nucleotide sequence comprising at least one AAV inverted terminal repeat (ITR) nucleotide sequence; a third nucleotide sequence comprising a Rep52 or a Rep40 coding sequence operably linked to expression control sequences for expression in an insect cell; and, a fourth nucleotide sequence comprising a Rep78 or a Rep68 coding sequence operably linked to expression control sequences for expression in an insect cell. The invention further relates to adeno-associated viral vectors with an altered ratio of the viral capsid proteins that provides improved infectivity of the viral particles.

Compositions and methods are provided for preparation of concentrated stock solutions of AAV virions without aggregation. Formulations for AAV preparation and storage are high ionic strength solutions (e.g. μ˜500 mM) that are nonetheless isotonic with the intended target tissue. This combination of high ionic strength and modest osmolarity is achieved using salts of high valency, such as sodium citrate. AAV stock solutions up to 6.4×1013 vg / mL are possible using the formulations of the invention, with no aggregation being observed even after ten freeze-thaw cycles. The surfactant Pluronic® F68 may be added at 0.001% to prevent losses of virions to surfaces during handling. Virion preparations can also be treated with nucleases to eliminate small nucleic acid strands on virions surfaces that exacerbate aggregation.

Described are the crystal structure of the alpha-helical domain of the gp41 component of HIV-1 envelope glycoprotein which represents the core of fusion-active gp41, methods of identifying and designing drugs which inhibit gp41 function and drugs which do so.

The subject invention relates to viruses that are able to purge (reduce or eliminate) undesirable cells in a mixture of cells. Undesirable cells can include neoplastic cells, cells mediating graft-versus host diseases, and autoimmune cells. The subject invention also relates to the purging of undesirable cells from bone marrow or peripheral blood cell harvests in the treatment of mammals including cancer patients, transplant recipients, and patients with autoimmune disease.

The present invention addresses the need to improve the long-term storage stability (i.e. infectivity) of vector formulations. In particular, it has been demonstrated that for adenovirus, the use of bulking agents, cryoprotectants and lyoprotectants imparts desired properties that allow both lyophilized and liquid adenovirus formulations to be stored at 4° C. for up to 6 months and retain an infectivity between 60–100% of the starting infectivity.

The present invention relates to viruses that are engineered to contain a surface ligand molecule which targets the virus to a cell of interest. In particular non-limiting embodiments, the cell of interest is desirably ablated and may be a cancer cell, an infected cell, a cell exhibiting a non-malignant proliferative disorder, or a cell of the immune system. Alternatively, the cell of interest is a target for gene therapy.

The present invention provides isolated promoters, transgene expression cassettes, vectors, kits, and methods for treatment of genetic diseases that affect the cone cells of the retina.

The present invention relates to novel adenovirus strains with an improved sero-prevalence. In one aspect, the present invention relates to isolated polypeptides of adenoviral capsid proteins such as hexon, penton and fiber protein and fragments thereof and polynucleotides encoding the same. Also provided is a vector comprising the isolated polynucleotide according to the invention and adenoviruses comprising the isolated polynucleotides or polypeptides according to the invention and a pharmaceutical composition comprising said vector, adenovirus, polypeptide and / or polynucleotide. The invention also relates to the use of the isolated polynucleotides, the isolated polypeptides, the vector, the adenoviruses and / or the pharmaceutical composition for the therapy or prophylaxis of a disease.

Substantially avirulent forms of atypical porcine reproductive and respiratory syndrome (PRRS) virus and corresponding vaccines are provided which result from cell culture passaging of virulent forms of PRRS. The resultant avirulent atypical PRRS virus is useful as a vaccine in that PRRS specific antibody response is elicited by inoculation of host animals, thereby conferring effective immunity against both previously known strains of PRRS virus and newly isolated atypical PRRS virus strains. The preferred passaging technique ensures that the virus remains in a logarithmic growth phase substantially throughout the process, which minimizes the time required to achieve attenuation. The present invention also provides diagnostic testing methods which can differentiate between animals infected with field strains and attenuated strains of PRRSV.

Disclosed are capsid-modified rAAV expression vectors, as well as infectious virions, compositions, and pharmaceutical formulations that include them. Also disclosed are methods of preparing and using novel capsid-protein-mutated rAAV vector constructs in a variety of diagnostic and therapeutic applications including, inter alia, as delivery agents for diagnosis, treatment, or amelioration of one or more diseases, disorders, or dysfunctions of the mammalian eye. Also disclosed are methods for intravitreal delivery of therapeutic gene constructs to retinal neuron cells, and specifically to ON bipolar cells, of the mammalian eye, as well as use of the disclosed compositions in the manufacture of medicaments for a variety of in vitro and / or in vivo applications including the treatment of retinitis pigmentosa, melanoma-associated retinopathy, and congenital stationary night blindness.

Compositions and methods are provided for preparation of concentrated stock solutions of AAV virions without aggregation. Formulations for AAV preparation and storage are high ionic strength solutions (e.g. μ˜500 mM) that are nonetheless isotonic with the intended target tissue. This combination of high ionic strength and modest osmolarity is achieved using salts of high valency, such as sodium citrate. AAV stock solutions up to 6.4×1013 vg / mL are possible using the formulations of the invention, with no aggregation being observed even after ten freeze-thaw cycles. The surfactant Pluronic® F68 may be added at 0.001% to prevent losses of virions to surfaces during handling. Virion preparations can also be treated with nucleases to eliminate small nucleic acid strands on virions surfaces that exacerbate aggregation.

The present invention provides cell lines for the production of E1-deleted adenovirus (rAd) vectors that complement E1A and E1B functions. The present invention also provides cell lines for the production of E1- and E2-deleted adenovirus vectors that complement E1A, E1B and E2B polymerase functions. The invention provides particular cell lines that complement E1A function by insertion of an E1A sequence containing mutations in the 243R and 289R proteins and an E1B sequence comprising the E1B-55K gene. Production yields in the resulting producer cell lines, designated SL0003 and SL0006, were similar to those obtained from 293 cells without generation of detectable recombinant replication competent adenovirus (“RCA”).

Methods to improve the tropism or other features of a virus are disclosed. Such methods can be used to prepare, e.g., DNA or plasmid libraries of variants of a gene encoding a viral capsid or envelope protein having a randomly inserted restriction site, libraries of viral clones with such variant genes with a randomly inserted restriction site or polypeptide sequence targeting a receptor expressed by a specific type of mammalian cells. Described are also methods to prepare mosaic viruses, i.e., viral particles wherein copies of one or more capsid or envelope proteins originate from different sources. These methods can be used to prepare mosaic viruses of a specific mixture of wild-type and mutant proteins, or of different types of mutant proteins.

A modified adenovirus capable of overcoming the problem of low level of coxsackie-adenovirus receptor (CAR) expression on tumor cells and methods of using such adenovirus are provided. The fiber protein of the adenovirus is modified by insertion or replacement so as to target the adenovirus to tumor cells, and the replication of the modified adenovirus is limited to tumor cells due to specific promoter control or mutations in E1a or E1b genes.

The present invention relates to gene therapy for tumors, specifically, it relates to the construction of oncolytic adenovirus recombinant, which specifically expresses immune modulatory factor GM-CSF in tumor cells and uses thereof.

The invention is related to an oncolytic adenovirus that comprises a sequence encoding a hyaluronidase enzyme inserted in its genome. This adenovirus spreads more efficiently in the tumour mass and therefore the oncolytic effect is increased. Injecting the oncolytic adenovirus of the invention endovenously, tumour volume regressions are obtained. Therefore the oncolytic adenovirus of the present invention is useful for the treatment of a cancer or a pre-malignant state of cancer.

The invention belongs to the technical field of biological control of pests and relates to a Lecanicillium lecanii and application of the Lecanicillium lecanii to control of greenhouse pests. The strain is Lecanicillium lecanii, is preserved in CGMCC (China General Microbiological Culture Collection Center) with the preservation number CGMCC No.8453, can be used for controlling greenhouse pests like whiteflies, aphids and thrips, and can be used in pollution-free vegetable and green food vegetable production greenhouses.

Methods to improve the tropism or other features of a virus is disclosed. Such methods can be used to prepare, e.g., DNA or plasmid libraries of variants of a gene encoding a viral capsid or envelope restriction site, libraries of viral clones with such variant genes with a randon-dy inserted restriction site or polypeptide sequence targeting a receptor expressed by a specific type of mammalian cells. Described are also methods to prepare mosaic viruses, i.e., viral particles wherein copies of one or more capsid or envelope proteins originate from different sources. These methods can be used to prepare mosaic viruses of a specific mixture of wild-type and mutant proteins, or of different types of mutant proteins.

A method for treating biological tissue, particularly meats for human consumption, so as to sanitize the tissue, is described. The method inactivates microorganisms and pathogenic prions (proteins) in the tissue.

The invention discloses culture media for edible mycorrhizal fungi and symbiotic seedlingsand and a synchronous culture method by utilizing the culture media. The culture media for the edible mycorrhizal fungi and the symbiotic seedlings comprise an edible mycorrhizal fungus strain mycelium culture medium and an edible mycorrhizal fungus symbiosis seedling synthesis culture medium, and the ediblemycorrhizal fungus strain mycelium culture medium includes a solid culture medium used for expanding reproduction of the mycelium and a liquid culture medium used for culturing mycelium which is usedfor inoculation of the mycorrhizal symbiotic seedlings. The synchronous culture method for implementing edible mycorrhizal fungus symbiotic seedling synthesis by utilizing the above culture media comprises the following steps: (1) a cultivation method for the edible mycorrhizal fungus strain mycelium, (2) a cultivation method for the edible mycorrhizal fungus symbiotic seedlings. According to theinvention, cultivation of the edible mycorrhizal fungus mycelium is divided into two stages including solid culture and liquid culture, liquid fungus strains are adopted for inoculation, and meanwhilethe edible mycorrhizal fungus mycelium and symbiotic plants in the mycorrhizal seedling synthesis culture medium grow together, so that the infection ability of the mycelium is increased by 30% or more, the success rate of the inoculation is increased by 20%-30%, and the survival rates of symbiotic plant seedlings and the mycorrhizal fungus mycelium are both increased by 20% or more.

The present inventors have developed a culture system for genotype 3a, which has a high prevalence worldwide. Since intergenotypic recombinant genomes exploiting the replication characteristics of JFH1 will be a valuable tool for the genotype specific study of the replaced genes and related therapeutics, the present inventors constructed a genotype 3a / 2a (S52 / JFH1) recombinant containing the structural genes (Core, E1, E2), p7 and NS2 of strain S52 and characterized it in Huh7.5 cells. S52 / JFH1 and J6 / JFH viruses passaged in cell culture had comparable growth kinetics and yielded similar peak HCV RNA titers and infectivity titers. Direct genome sequencing of cell culture derived S52 / JFH1 viruses identified putative adaptive mutations in Core, E2, p7, NS3 and NS5A; clonal analysis revealed, that all genomes analyzed exhibited different combinations of these mutations. Finally, viruses resulting from transfection with RNA transcripts of five S52 / JFH1 recombinant containing these combinations of putative adaptive mutations performed as efficiently as J6 / JFH viruses in Huh7.5 15 cells and were all genetically stable after viral passage. In conclusion, the present inventors have developed a robust and genetically stable cell culture system for HCV genotype 3a.

The present invention addresses the need to improve the long-term storage stability (i.e. infectivity) of vector formulations. In particular, it has been demonstrated that for adenovirus, the use of bulking agents, cryoprotectants and lyoprotectants imparts desired properties that allow both lyophilized and liquid adenovirus formulations to be stored at 4° C. for up to 6 months and retain an infectivity between 60-100% of the starting infectivity.

The subject invention relates to viruses that are able to purge (reduce or eliminate) undesirable cells in a mixture of cells. Undesirable cells can include neoplastic cells, cells mediating graft-versus host diseases, and autoimmune cells. The subject invention also relates to the purging of undesirable cells from bone marrow or peripheral blood cell harvests in the treatment of mammals including cancer patients, transplant recipients, and patients with autoimmune disease.

The invention relates to phage lyase composite powder. The composite powder composition comprises the following components in percentage by mass: 0.15%-0.2% of phage lyase TSPpgh, 0.15%-0.2% of phagelyase MMPpgh, 0.3%-0.5% of trehalose, 0.1%-0.2% of citric acid, 0.2%-0.3% of mannitol, 0.2%-0.3% of vitamin C and 98.3%-98.9% of diatomite. The phage lyase composite powder is subjected to in-vitro antibacterial activity detection by adopting a two layer plating method. After the composite powder is stored for 12 months at normal temperature, the enzyme activity is kept at 95% or above (the enzymeactivity loss is within 5%). The composite lyase preparation can be used for replacing antibiotics to inhibit proliferation of harmful pathogenic bacteria such as escherichia coli, salmonella and staphylococcus aureus in animal intestines, and can maintain intestinal microecological balance.

The present invention relates to novel adenovirus strains with an improved seroprevalence. In one aspect, the present invention relates to isolated polypeptides of adenoviral capsid proteins such as hexon, penton and fiber protein and fragments thereof and polynucleotides encoding the same. Also provided is a vector comprising the isolated polynucleotide according to the invention and adenoviruses comprising the isolated polynucleotides or polypeptides according to the invention and a pharmaceutical composition comprising said vector, adenovirus, polypeptide and / or polynucleotide. The invention also relates to the use of the isolated polynucleotides, the isolated polypeptides, the vector, the adenoviruses and / or the pharmaceutical composition for the therapy or prophylaxis of a disease.

The invention provides a method for conjugating a peptide displayed on a genetic display system to a molecular scaffold performed on an ion exchange resin.

A modified adenovirus capable of overcoming the problem of low level of coxsackie-adenovirus receptor (CAR) expression on tumor cells and methods of using such adenovirus are provided. The fiber protein of the adenovirus is modified by insertion or replacement so as to target the adenovirus to tumor cells, and the replication of the modified adenovirus is limited to tumor cells due to specific promoter control or mutations in E1a or E1b genes.