60results about "Invertebrate cells" patented technology

The invention discloses a preparation method and application of a sea cucumber exosome with an immune cell repairing function, and belongs to the technical field of biomedicine. The method comprises the following steps: feeding sea cucumbers by using a combined method of composite stress and an echinacea extract, extracting sea cucumber cells, and obtaining sea cucumber exosomes through ultracentrifugation; wherein the condition of composite stress is set as follows: the day and night temperature difference is not lower than 3 DEG C, and meanwhile, the dissolved oxygen content of the water body is adjusted to 75-90% of the dissolved oxygen content of the conventional feeding water body. The obtained sea cucumber exosome has a remarkable immune cell repairing function, and can effectively promote proliferation and activity recovery of immune cells, inhibit expression of inflammatory factors and improve the level of anti-inflammatory factors. The sea cucumber exosome prepared by the preparation method disclosed by the invention has an important application prospect in the research and development of immune repair drugs, and can provide a technical support for a novel immune regulation preparation in the field of biomedicine.

The invention discloses a method suitable for extracting extracellular small vesicles in fresh water pearl shell tissue waste liquid, which comprises the following operation steps: (1) solid-liquid separation: slaughtering pearl shells, and filling slaughtering waste liquid into a waste liquid crude extraction device; (2) waste liquid crude extraction: performing multi-stage gradient filtration and gravity settling on the slaughtering waste liquid by using a waste liquid crude extraction device to obtain clarified waste liquid; (3) removing impurities: removing impurities for the first time and removing impurities for the second time; (4) concentration of the extracellular vesicles: concentrating the extracellular vesicles by using an ultra-speed centrifuge to obtain a high-concentration extracellular vesicle solution; and (5) preservation of extracellular vesicles: according to the final volume of the high-concentration extracellular vesicle solution obtained in the step (4), carrying out sample standardization through protein concentration, sub-packaging the sample, and carrying out long-term preservation in a low-temperature environment. The particle sizes of the extracted vesicles are concentrated, high-purity and complete extracellular small vesicles can be obtained, and the method is suitable for biomedical research and product development.

The invention relates to the technical field of bioengineering, in particular to an exosome separation and concentration method with marine animal source tissue as a raw material. Comprising the following steps: freeze-drying and crushing a marine animal source tissue sample; performing sample enzymolysis; and removing impurities and concentrating. According to the method disclosed by the invention, the exosome in the cells is fully dissociated through freeze-dried powder crushing and composite enzymolysis processes, and the filtering speed of the sample is accelerated by virtue of ultrasonic oscillation, so that the rapid and high-purity separation and concentration of the exosome in the marine animal source tissue are realized.

The invention discloses a culture method of in-vitro uniform monolayer cells of prawns, and belongs to the technical field of biology. The method comprises the following steps: (1) obtaining lymphatic tissues of a last-stage prawn at the early stage of shelling, completely stripping an outer membrane to obtain internal tissues, and cutting the internal tissues into small tissue blocks; (2) cleaning the small tissue blocks, inoculating the small tissue blocks to a culture plate coated with a gelatin solution, adding a complete culture solution into an inoculation system at intervals within 12 hours after inoculation for culture, replacing the complete culture solution 24-48 hours after inoculation, and inducing tissue cells to continuously migrate to form an in-vitro cell layer; (3) blowing and beating the cell culture to obtain a cell suspension, and centrifuging to obtain a cell monolayer; and (4) suspending the cell monolayer by adopting a complete culture solution, then inoculating the cell monolayer into a culture plate coated with a gelatin solution, and culturing to obtain the in-vitro uniform monolayer cells of the prawns. According to the method, the prawn monolayer cells which are long in in-vitro survival time, uniform and stable and are cultured in vitro can be obtained, and an important tool is provided for related virology research.

Provided is a detection technique excellent in cell drying resistance, cell retention, and detection of a chemical substance such as an odorant. More specifically, provided are: a laminate containing

The present invention provides a method for the production of a recombinant adeno-associated viral vector (rAAV), comprising contacting a host cell with a solution comprising at least one compound of formula (I), (IA), (IB), (II), (III), or (IV), or a salt thereof, or vitamin B, or any combination thereof. Also provided is a method for increasing production of rAAV by a host cell, comprising contacting the host cell with a solution comprising at least one compound of formula (I), (IA), (IB), (II), (III), or (IV), or a salt thereof, or vitamin B, or any combination thereof.

Provided are non-plant polypeptides or proteins comprising xylosylated N-linked glycans containing β1,2-xylose, wherein at least 25% of the individual protein species comprise xylosylated N-glycans, compositions comprising the non-plant proteins, cells engineered to produce the polypeptides or proteins, and methods for their production and use.

The invention relates to the technical field of biology, and discloses a method for constructing a drosophila embryo permeabilization model, which comprises the following steps: collecting and cleaning drosophila embryos: collecting and washing the drosophila embryos; rinsing the drosophila melanogaster embryos, namely rinsing the drosophila melanogaster embryos by using a bleaching agent solution to remove chorion layers of the drosophila melanogaster embryos, and washing the embryos after rinsing; and performing permeation treatment on the drosophila melanogaster embryo: performing permeation treatment on the drosophila melanogaster embryo without the chorion layer in an embryo permeation solution EPS, and then cleaning in BPS to obtain the drosophila melanogaster embryo permeation model. According to the construction method of the drosophila melanogaster embryo permeabilization model, operation is easy, convenient and rapid, it can be guaranteed that embryo permeability is good, embryo vitality is maintained at the same time, the influence of small molecule drugs on the embryo development process is conveniently studied, and the method is suitable for popularization and application. And favorable conditions are provided for researching cell and molecular mechanisms of embryonic development, toxicological evaluation of small molecular substance screening and screening of related drugs in an embryo model.

Provided are an engineered mRNA and a method for improving protein expression using same. Also provided is the use of such engineered mRNA in gene therapy and / or genetic vaccination.

The present disclosure provides a method for producing a methyl compound using glycine, serine, or an organic raw material.

The present disclosure provides a polypeptide fragment comprising a polypeptide of a modified hemagglutinin protein, wherein said modified hemagglutinin protein comprises a linker peptide replacing the polypeptide in the cytoplasmic domain of hemagglutinin protein, wherein the hemagglutinin protein is obtained from at least one seasonal strain of influenza virus, wherein said polypeptide of the modified hemagglutinin protein is having at least 80%, or at least 90%, or at least 95%, or at least 97% sequence identity to at least one sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, and SEQ ID NO: 30.

Provided are a recombinant adeno-associated virus (rAAV) genome and a single-polarity rAAV (spAAV) vector formed by packaging same. Specifically, provided is an spAAV genome before packaging, two ends of which are both linear open terminuses, wherein one end does not have an inverted terminal repeat (ITR) or a part thereof, and the other end has a truncated ITR (ITRT), and the ITRT does not form a T-shaped hairpin palindromic structure. The spAAV genome is a single-polarity single-stranded DNA. Also provided are an spAAV vector, a method for delivering an exogenous nucleic acid to a cell by using the vector, and a use of the vector in the preparation of a product for gene expression, gene therapy, gene editing or gene regulation.

The present invention relates to the use of parasites and extracellular vesicles obtained from parasites for cancer treatment. The aim of the present invention is to use parasites and extracellular vesicles obtained from parasites in cancer treatment and to load active substances onto the exosomes by using the drug loading capacity of the exosomes and thus to carry the specific drug directly to the target cancer cells without causing any side effects on healthy cells and thereby to increase the bioavailability of the drug to achieve the desired effect in the tumor-specific target area. Within the scope of the present invention, in particular, the infantile leishmania parasite is used as a source of extracellular vesicles.

This invention belongs to the field of genetic engineering, specifically relating to the application of RSAD1 gene / protein as a target in screening drugs to inhibit infection by microribonucleoviridae viruses. This invention unexpectedly discovered that silencing the RSAD1 gene can significantly inhibit Seneca virus replication, and can be used as a target for the development of antiviral drugs against Seneca virus. This invention provides an sgRNA sequence targeting the RSAD1 gene, and combined with CRISPR-Cas9 gene editing technology, silences the RSAD1 gene, obtaining an RSAD1 gene knockout monoclonal cell line. This cell line can significantly inhibit Seneca virus replication, providing a target for further screening of drugs and reagents to resist Seneca virus replication. Simultaneously, this invention found that overexpression of the RSAD1 protein in host cells can significantly promote Seneca virus replication, and can be used as a production cell line for Seneca virus or vaccines.

The present invention provides a method of releasing viral vectors from cells producing those viral vectors by contacting the cells with a photosensitising agent which is then irradiated to disrupt the plasma membrane of the cells to release the viral vectors which may be collected and / or purified. The product of such methods as well as kits and apparatuses for performing the methods are also provided.

The application provides a Marburg GP protein mutant, a DNA molecule, a recombinant vector and application thereof, and belongs to the technical field of Marburg virus vaccine preparation. The Marburg GP protein mutant is obtained by site mutation on the basis of a wild-type Marburg GP protein, and contains at least one mutation selected from R575P, V576P and L585P. The amino acid sequence of the wild-type Marburg GP protein is shown as SEQ ID NO:1. The Marburg GP mutant can provide a stable GP pre-fusion conformation, and can induce stronger binding activity, neutralizing activity and protective antibodies in mice, and is expected to be used for preparation of a Marburg virus vaccine.

To provide an adeno-associated virus vector (AAV) that can be very efficiently produced in insect cells.SOLUTION: The invention provides a nucleic acid construct comprising expression control sequences for expression in an insect cell of a nucleotide sequence comprising an open reading frame, the open reading frame sequence encodes: i) AAV capsid proteins VP1, VP2 and VP3; and ii) an ATG translation initiation codon for VP1. The nucleotide sequence also comprises, upstream of the open reading frame, an alternative start codon which is out of frame with the open reading frame.SELECTED DRAWING: None

To provide recombinantly-modified adeno-associated virus (AAV) helper vectors that are capable of increasing the packaging efficiency of recombinantly-modified adeno-associated virus (rAAV) and their use to improve the packaging efficiency of such rAAV.SOLUTION: The present invention is directed to recombinantly-modified adeno-associated virus (AAV) helper vectors that have been further modified to replace (or augment) P5 and / or P40 promoter sequences that are natively associated with Rep proteins encoded by rAAV with AAV P5 and / or P40 promoters that are associated with the Rep proteins of an rAAV of different serotype.SELECTED DRAWING: Figure 1

The invention discloses a yellow thorn moth blood lymphocyte line, an expanded production method of a nuclear polyhedrosis virus and application of the yellow thorn moth blood lymphocyte line and the nuclear polyhedrosis virus. The name of the dalbergia mandshurica blood lymphocyte line is dalbergia mandshurica blood lymphocyte CFW-CHY, the dalbergia mandshurica blood lymphocyte line is preserved in the China Center for Type Culture Collection (CCTCC), the preservation number is CCTCC NO: C2025104, and the preservation date is March 27, 2025; the yellow thorn moth blood lymphocyte line is used for expanded production of the nuclear polyhedrosis virus, so that industrial production of the nuclear polyhedrosis virus is realized, and the yellow thorn moth blood lymphocyte line can be used in the fields of preparation of insecticides, insecticidal pesticides and the like.

The invention provides a catechol-O-methyltransferase mutant as well as a preparation method and application thereof, and relates to the technical field of biology. The catechol-O-methyltransferase mutant contains 79P mutation relative to wild type catechol-O-methyltransferase, the catechol-O-methyltransferase mutant can be expressed in vitro through recombinant cells, and the problems that in the prior art, a catechol-O-methyltransferase preparation process is poor in stability and high in cost are solved.

The invention discloses a recombinant expression and purification method of SLFN14 protein. The invention provides a method for improving the expression quantity and / or the stability of SLFN14 protein, which comprises the following steps of: A1) mutating cysteine at 365th, 775th and 808th sites of an amino acid sequence of the SLFN14 protein into serine, and mutating glutamic acid at 211th site into alanine to obtain an SLFN14 protein mutant; a2) expressing the SLFN14 protein mutant through a baculovirus-insect cell system, so as to obtain an insect cell for expressing the SLFN14 protein mutant; a3, the insect cells expressing the SLFN14 protein mutant are subjected to splitting decomposition, nickel column purification, ion exchange purification and molecular sieve purification in sequence, and the target protein.The purity, yield and stability of SLFN14 are remarkably improved through the method.

The invention relates to the technical field of biology, in particular to hyriopsis cumingii exosome-like nano vesicles and application thereof in preparation of osteogenesis products. According to the preparation method, hyriopsis cumingii is taken as a raw material, and the hyriopsis cumingii exosome-like nano-vesicles are obtained through separation and purification by adopting a differential centrifugation and ultracentrifugation combined method; the nano-vesicle has a complete membrane structure and typical exosome-like morphological characteristics, in-vitro experiments prove that the nano-vesicle can significantly promote proliferation of mouse cranial preosteoblasts (MC3T3-E1) and shows osteogenic activity, and a new way is provided for development of aquatic product-derived osteogenic functional factors.

A baculovirus vector and a use thereof in the preparation of a recombinant adeno-associated virus (rAAV) in an insect cell are provided. The baculovirus vector includes an exogenous gene expression cassette and a stable sequence. The stable sequence is located at a site 5 kb or less from the exogenous gene expression cassette, and the stable sequence is a conserved noncoding element (CNE) sequence or a nucleocapsid assembly-essential element (NAE) sequence. When an insect cell is infected with a recombinant baculovirus (rBV) constructed in this way, after multiple continuous passages, production levels of the rBV and the rAAV still remain relatively stable.

Provided is a method for producing a protein having excellent expression efficiency, in which a baculovirus expression system is used.　The method for producing a protein using a baculovirus expression system comprises a step for inoculating recombinant baculovirus into insect cells and then adding a calcium salt to a culture medium.

This invention discloses a human metapneumovirus virus-like particle, its preparation method, its application, and a vaccine. The human metapneumovirus virus-like particle is assembled from the M1 protein of influenza virus H2N2 and the F protein of human metapneumovirus. The nucleic acid sequence encoding the M1 protein is shown in SEQ ID NO:1, and the nucleic acid sequence encoding the F protein is shown in SEQ ID NO:2. The preparation method includes (1) constructing a recombinant plasmid: cloning the M1 gene of influenza virus H2N2 and the F gene sequence of human metapneumovirus into the pFastBacDual vector to obtain the recombinant plasmid pFastBacDual-M1-F; (2) transfection: transfecting the recombinant plasmid pFastBacDual-M1-F into sf9 insect cells to obtain recombinant sf9 cells; (3) isolation and purification: culturing the recombinant sf9 cells and centrifuging to purify and obtain human metapneumovirus virus-like particles (VLPs). This invention uses a baculovirus insect cell expression system to prepare virus-like particles based on HMPV F protein and influenza virus M1 protein, which have immunogenicity and excellent immunoprotective effects, providing theoretical data and laying the foundation for the development of HMPV vaccines, especially VLP vaccines.

Provided are: a method for culturing animal cells, wherein animal cells are cultured in the presence of an iron ion carrier such as hinokitiol; and a technique related thereto.

The present invention relates to a cell culture medium which sustains the growth of cells obtained from a crustacean in vitro. Indeed, the present invention discloses a cell culture medium comprising a specific salt composition, in part responsible for the correct osmolality and pH, a specific amino acid and vitamin composition, energy sources such as glucose, and crustacean hemolymph. The present invention further discloses that said cell culture medium is capable to keep cells obtained from shrimp and lobster viable for long periods of time even after passaging the cells into subcultures and induces said cells to proliferate. The cultured cells are for example useful to perform physiological studies on certain genes (by genetic engineering) and to do research and diagnosis on crustacean pathogens.

Described herein are methods for manufacturing viruses and viral particles. In some aspects, the methods use a sugar-based detergent for lysing a host cell to release the virus or viral particle. Also, described herein is a composition or a pharmaceutical composition comprising the virus or viral particle manufactured by the methods described herein.

The present invention provides a method of manufacturing a coral scaffold for use as a bone graft substitute. The method comprises growing coral in a growth medium having a carbonate hardness, dKH, of 10 or more; removing at least a portion of the coral from the growth medium; devitalising coral removed from the growth medium and sizing the devitalised coral to form the coral scaffold.

An object of the present invention is to provide a novel method that can produce a methyl compound by regenerating SAM and accelerating the methylation reaction of a compound. The present invention provides a method for producing a methyl compound using glycine, serine, or an organic raw material.