96 results about "Cold adapted" patented technology

The cold-adapted master strain A / Ann Arbor / 6 / 60 7PI (H2N2) and progenitor wild type E2(3) viral strains have been deposited and their genomic sequences identified. Seven nucleotide differences were found between the sequences identified herein and the previously published sequences for cold-adapted A / Ann Arbor / 6 / 60 genes. The cold-adapted live influenza virus of the present invention can be reassorted with a variety of epidemic wild type influenza viruses and used to produce vaccines to prophylactically and therapeutically treat influenza.

Chimeric respiratory syncytial virus (RSV) and vaccine compositions thereof are produced by introducing one or more heterologous gene(s) or gene segment(s) from one RSV subgroup or strain into a recipient RSV backround of a different subgroup or strain. The resulting chimeric RSV virus or subviral particle is infectious and attenuated, preferably by introduction of selected mutations specifying attenuated phenotypes into a chimeric genome or antigenome to yield, for example, temperature sensitive (ts) and / or cold adapted (ca) vaccine strains. Alternatively, chimeric RSV and vaccine compositions thereof incorporate other mutations specifying desired structural and / or phenotypic characteristics in an infectious chimeric RSV. Such chimeric RSV incorporate desired mutations specified by insertion, deletion, substitution or rearrangement of one or more selected nucleotide sequence(s), gene(s), or gene segment(s) in a chimeric RSV clone. This provides a method for development of novel vaccines against diverse RSV strains by using a common attenuated backbone as a vector to express protective antigens of heterologous strains. The immune system of an individual is stimulated to induce protection against natural RSV infection, preferably in a multivalent manner to achieve protection against multiple RSV strains and / or subgroups.

The present invention provides experimentally-generated cold-adapted equine influenza viruses, and reassortant influenza A viruses comprising at least one genome segment of such an equine influenza virus, wherein the equine influenza virus genome segment confers at least one identifying phenotype of the cold-adapted equine influenza virus, such as cold-adaptation, temperature sensitivity, dominant interference, or attenuation. Such viruses are formulated into therapeutic compositions to protect animals from diseases caused by influenza A viruses, and in particular, to protect horses from disease caused by equine influenza virus. The present invention also includes methods to protect animals from diseases caused by influenza A virus utilizing the claimed therapeutic compositions. Such methods include using a therapeutic composition as a vaccine to generate a protective immune response in an animal prior to exposure to a virulent virus, and using a therapeutic composition as a treatment for an animal that has been recently infected with a virulent virus, or is likely to be subsequently exposed to virulent virus in a few days whereby the therapeutic composition interferes with the growth of the virulent virus, even in the absence of immunity. The present invention also provides methods to produce cold-adapted equine influenza viruses, and reassortant influenza A viruses having at least one genome segment of an equine influenza virus generated by cold-adaption.

Vectors and methods for the production of influenza viruses suitable as recombinant influenza vaccines in cell culture are provided. Bi-directional expression vectors for use in a multi-plasmid influenza virus expression system are provided. Additionally, the invention provides methods of producing influenza viruses with enhanced ability to replicate in embryonated chicken eggs and / or cells (e.g., Vero and / or MDCK) and further provides influenza viruses with enhanced replication characteristics. A method of producing a cold adapted (ca) influenza virus that replicates efficiently at, e.g., 25° C. (and immunogenic compositions comprising the same) is also provided.

The invention relates to an immobilized cold-adapted microorganismmicrobe carbonized sludge carrier filler for sewage treatment and application thereof. According to the filler, carbonized sludge, polyvinyl alcohol, sodium alginate and concentrated carrageenan powder are optimally proportioned; mixed with the concentrated carrageenan powder, activated sludge biochar as an adsorbing carrier mixed with three strains of pyschrophiles, and with polyvinyl alcohol, sodium alginate and the concentrated carrageenan powder as embedding carriers and calcium chloride as a cross-linking agent, the carbonized sludge carrier filler with enriched immobilized cold-adapted microbes is produced. When the microorganismmicrobe filler disclosed by the invention is applied into the intensified treatment of winter sewage, the removal rates of ammonia nitrogen, TN, TP and COD in water can reach 80.09 to 85.27 percent, 76.24 to 83.67 percent, 79.72 to 85.05 percent and 85.01 to 90.52 percent, and the method disclosed by the invention is an energy-saving and high-efficiency biological intensified remedication technique.

The present invention provides methods for the purification of cell-associated viruses from adherent cells (e.g., MDCK or Vero cells). In particular, the present invention provides purification methods for the production of immunogenic compositions comprising a live attenuated cell-associated virus (e.g., an attenuated respiratory syncytial virus (RSV) or cold-adapted, and / or temperature sensitive influenza virus) that result in levels of host cell DNA (HCD), host cell protein (HCP) and non-specific endonuclease (e.g., Benzonase), which are below the specifications required by regulatory agencies. The immunogenic compositions can be used to actively immunize subjects or to generate antibodies for a variety of uses, including passive immunization and diagnostic immunoassays.

The present invention provides experimentally-generated cold-adapted equine influenza viruses, and reassortant influenza A viruses comprising at least one genome segment of such an equine influenza virus, wherein the equine influenza virus genome segment confers at least one identifying phenotype of the cold-adapted equine influenza virus, such as cold-adaptation, temperature sensitivity, dominant interference, or attenuation. Such viruses are formulated into therapeutic compositions to protect animals from diseases caused by influenza A viruses, and in particular, to protect horses from disease caused by equine influenza virus. The present invention also includes methods to protect animals from diseases caused by influenza A virus or other infectious agents utilizing the claimed therapeutic compositions. Such methods include using a therapeutic composition as a vaccine to generate a protective immune response in an animal prior to exposure to an infectious agent, as well as using a therapeutic composition as a treatment for an animal that has been recently infected with an infectious agent leading to respiratory disease, or is likely to be subsequently exposed to such an agent in a few days whereby the therapeutic composition reduces such respiratory disease, even in the absence of antibody-mediated immunity. The present invention also provides methods to produce cold-adapted equine influenza viruses, and reassortant influenza A viruses having at least one genome segment of an equine influenza virus generated by cold-adaptation.

Attenuated respiratory syncytial virus (RSV) and vaccine compositions thereof are produced by introducing specific mutations associated with attenuating phenotypes into wild-type or RSV which is incompletely attenuated by cold-passage or introduction of mutations which produce virus having a temperature sensitive (ts) or cold adapted (ca) phenotype. Alternatively, recombinant RSV and vaccine compositions thereof incorporate attenuating and other mutations specifying desired structural and or phenotypic characteristics in an infectious RSV. Recombinant RSV incorporate desired mutations specified by insertion, deletion, substitution or rearrangement of a selected nucleotide sequence, gene, or gene segment in an infectious RSV clone. The immune system of an individual is stimulated to induce protection against natural RSV infection, or multivalently against infection by RSV and another pathogen, such as PIV, by administration of attenuated, biologically derived or recombinant RSV.

The invention provides an extraction method of bovine tendon collagen. The extraction method has the advantages of high yield, short production period and high product purity, and comprises the following steps of: preprocessing bovine tendon, and freezing and crushing the bovine tendon; putting the crushed bovine tendon into a Tris-HCL buffer solution; then, adding cold-adapted protease MCP-01 accounting for 0.1 percent of the weight of the bovine tendon into the buffer solution; performing enzymolysis at 40 DEG C; then, obtaining an enzymolysis solution; centrifuging the enzymolysis solution; taking precipitates; dissolving the precipitates by 0.1-percent acetic acid; performing filtration to obtain filter liquid; salting out the filter liquid to obtain a salting-out solution; performing centrifugation twice; adding water for dissolution; then, adding 0.1-percent acetic acid for continuous dissolution; and performing freeze drying to obtain the bovine tendon collagen. The extraction method has the beneficial effects that the cold-adapted protease MCP-01 is used for hydrolyzing the bovine tendon in a buffer liquid system, so that collagen helix molecules of collagen micro fiber are exposed; collagen filaments expand and loose, so that the hydrolysis is fast and sufficient; and the yield and the product purity are further improved.

The invention relates a gene for coding low-temperature alkaline lipase (LipY). The gene is obtained by cloning cold-adapted marine yeast (BoHai Sea-9145), which can secrete low-temperature alkaline lipase and is screened from sea mud of Bohai, by a method of combining degenerate primer polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE). The gene contains a 1206bp open reading frame (ORF), codes 401 amino acids and contains a signal peptide with 28 amino acids at the amino acid end. Each coded amino acid contains a conserved sequence (G-X-S-X-G) of the lipase and a potential N-glycosylation site. The lipase gene is cloned on an eukaryotic expression vector (pIc9K) and performs heterologous expression in Pichia pastoris (Gs115). High-activity recombinant lipase is obtained from supernate 96 hours after fermentation, wherein methanol serves as an inducer. An electrophoretic pure lipase sample is obtained by purifying the recombinant lipase by a nickel ion affinity column. The recombinant lipase has a series of advantages of high low-temperature catalytic activity, stable performance under the alkaline condition and the like, and has wide application prospect in the low-temperature catalytic fields of daily chemicals, textile, food processing and the like.

The present invention provides novel serum-free cell culture medium and methods for cultivating MDCK cells. In particular, non-tumorigenic MDCK cells. The present invention also provides methods for producing influenza viruses (e.g., particularly cold-adapted, and / or temperature sensitive, and / or attenuated influenza viruses) that eliminate the need for a cell culture medium exchange step. The novel medium and methods are useful to grow influenza viruses, in cell culture to high titer. The present invention further provides purification methods for purifying influenza viruses with high overall recovery of live virus and result in levels of host cell DNA (HCD), host cell protein (HCP) and non-specific endonuclease (e.g., Benzonase), which are below the specifications required by regulatory agencies. The immunogenic compositions can be used to actively immunize subjects or to generate antibodies for a variety of uses, including passive immunization and diagnostic immunoassays.

Attenuated respiratory syncytial virus (RSV) and vaccine compositions thereof are produced by introducing specific mutations associated with attenuating phenotypes into wild-type or RSV which is incompletely attenuated by cold-passage or introduction of mutations which produce virus having a temperature sensitive (ts) or cold adapted (ca) phenotype. Alternatively, recombinant RSV and vaccine compositions thereof incorporate attenuating and other mutations specifying desired structural and or phenotypic characteristics in an infectious RSV. Recombinant RSV incorporate desired mutations specified by insertion, deletion, substitution or rearrangement of a selected nucleotide sequence, gene, or gene segment in an infectious RSV clone. The immune system of an individual is stimulated to induce protection against natural RSV infection, or multivalently against infection by RSV and another pathogen, such as PIV, by administration of attenuated, biologically derived or recombinant RSV.

The present invention relates to an isolated attenuated influenza virus strain and a live vaccine comprising the same. The isolated attenuated influenza virus strain is prepared by cold-adaptation of a mother strain which carries 6 internal genomes of A / PR / 8 / 34(H1N1) and two surface antigens HA and NA of A / Aichi / 2 / 68(H3N2). The attenuated influenza virus strain and the live vaccine of the present invention are useful for prevention of seasonal influenza episodes and sudden outbreak of influenza pandemics of predicted or unknown identity, since they have safety, efficacy, high production yield, immediate protection against variety of influenza subtypes and prolonged protection against specific influenza subtype.

The invention relates to novel alginate lyase with cold-adapted characteristics and an application thereof. The alginate lyase is artificially-designed novel alginate lyase AlgA7y, an amino acid sequence is as shown by SEQ ID NO.1, an N-end N comprises a section of carbohydrate combination domain (Met1-Pro176), an C-end C comprises a section of alginate lyase preserved catalytic area (Ser177-Asp526), and the identity between the amino acid sequence of the alginate lyase in provided bythe invention and the alginate lyase with known functions is only 71 percent. The alginate lyase AlgA7 of provided by the invention has cold-adapted characteristics, the most appropriate reaction temperature is 25 DEG C, and the reaction activity at 20 to and 10 DEG C can reach 81.9 percent to and42.6 percentof the reaction activity at the most appropriate temperature. The alginate lyase can control the enzymatic reaction to be performed at a low temperature, so that the energy consumption in the industrialized application process can be saved, and the cost can be savedreduced.

The present invention provides experimentally-generated cold-adapted equine influenza viruses, and reassortant influenza A viruses comprising at least one genome segment of such an equine influenza virus, wherein the equine influenza virus genome segment confers at least one identifying phenotype of the cold-adapted equine influenza virus, such as cold-adaptation, temperature sensitivity, dominant interference, or attenuation. Such viruses are formulated into therapeutic compositions to protect animals from diseases caused by influenza A viruses, and in particular, to protect horses from disease caused by equine influenza virus. The present invention also includes methods to protect animals from diseases caused by influenza A virus utilizing the claimed therapeutic compositions. Such methods include using a therapeutic composition as a vaccine to generate a protective immune response in an animal prior to exposure to a virulent virus, and using a therapeutic composition as a treatment for an animal that has been recently infected with a virulent virus, or is likely to be subsequently exposed to virulent viruses in a few days whereby the therapeutic composition interferes with the growth of the virulent virus, even in the absence of immunity. The present invention also provides methods to produce cold-adapted equine influenza viruses, and reassortant influenza A viruses having at least one genome segment of an equine influenza virus generated by cold-adaptation.

The invention relates to cold-adapted cellulase-producing bacillus subtilis and a screening method thereof. The cold-adapted cellulase-producing bacillus subtilis comprises bacillus subtilis with a preservation mechanism being General Microbiological Culture Center for China Committee for Culture Collection of Microorganisms, a preservation number being CGMCC No. 18001 and a preservation date being June 19, 2019. For the cold-adapted cellulase-producing bacillus subtilis and the screening method, efficient cold-adapted cellulase bacillus subtilis is separated and screened from soil in different low-temperature areas, and the biological characteristics of the bacillus subtilis are as follows: a bacterial colony on an LB culture medium is relatively small, round, wet, smooth and sticky, is easy to pick and has typical bacterial colony characteristics; and in addition, the influences of the conditions, including temperature, carbon sources, nitrogen sources and a pH value, of the bacterial colony on cellulase are studied. By applying the cold-adapted cellulase-producing bacillus subtilis to straw returning in northeast low-temperature cold areas, the influence of straw incineration inthe northeast areas on environment is reduced, and a solid foundation is laid for sustainable development of agriculture.