141 results about "Cell lysates" patented technology

The present invention provides a Lactiplantibacillus plantarum ( Lactiplantibacillus plantarum ), strain 24, which is deposited in the China General Microbiological Culture Collection Center, deposit number: CGMCC No. 31278. The Lactiplantibacillus plantarum 24 of the present invention can secrete sphingomyelinase to promote the production of ceramide in HaCaT cells; Lactiplantibacillus plantarum 24 or its metabolite or its lysate can up-regulate the expression of key enzymes in the sphingomyelinase and / or ceramide synthesis pathway of skin cells, repair the skin barrier; down-regulate the expression of pro-inflammatory cytokines, inhibit autophagy, and has a good anti-inflammatory effect; at the same time, it also has good antioxidant and whitening effects. The Lactiplantibacillus plantarum 24 or its metabolite or its cell lysate of the present invention can be used in the preparation of products having the above-mentioned effects or functions.

The invention discloses a single-cell metabolite spectrum quantitative analysis platform and method based on droplet microfluidics, and relates to the technical field of mass spectrometry. The platform comprises a fluid pumping module, a cell oscillation module, a microfluidic mixed liquid drop generation module, a microfluidic mixed liquid drop reinjection and separation module and a mass spectrometry module. The fluid pumping module comprises a high-pressure nitrogen cylinder and a gas pressure driver; the cell oscillation module comprises a cell oscillator; the micro-fluidic mixed droplet generation module comprises a micro-fluidic droplet generation chip, a cell lysis solution, an internal standard, an oil phase, a cell suspension and a droplet collection and incubation centrifugal tube; the micro-fluidic mixed liquid drop reinjection and separation module comprises a micro-fluidic liquid drop reinjection and separation chip; and the mass spectrometry module comprises an electrospray needle and a mass spectrometer. According to the method, the single cell is captured and the internal standard substance is introduced at the same time by using droplet microfluidics, and mass spectrometry is carried out, so that high-flux single cell encapsulation and accurate quantification of metabolites are realized.

The invention discloses a moisturizer prepared from a lysate extract produced by fermentation of bacillus subtilis. The hydroxydecyl ubiquinone is identified in Y-3FLE, and the yield of the hydroxydecyl ubiquinone is 0.45 mu g / g after the hydroxydecyl ubiquinone is cultured for 4 days, so that the quinone substances are identified in the Halophilic bacteria in the Yuncheng Salt Lake for the first time. The Y-3FLE is prepared by adopting absolute ethyl alcohol as a solvent, the oxidation resistance of the Y-3FLE is investigated through an in-vitro experiment, and a result shows that the Y-3FLE is relatively high in reducing capacity and relatively high in oxidation resistance. A human body closed patch test result shows that the Y-3FLE has no obvious irritation to human skin. The method is applied to the moisturizer, and human body efficacy evaluation is carried out through a multi-probe skin tester. Results show that the moisturizer added with 1% of Y-3FLE has multiple remarkable effects of moisturizing, repairing, nourishing and the like.

The invention belongs to the field of biology, and discloses a method for identifying RNA (Ribonucleic Acid) binding protein in porcine intramuscular fat cells or subcutaneous fat cells, which comprises the following steps: step 1, taking porcine muscle tissues or subcutaneous fat tissues to carry out meat mashing and digestion, and collecting mature intramuscular fat cells or subcutaneous fat cells; step 2, washing intramuscular fat cells or subcutaneous fat cells, and carrying out cross-linking treatment by adopting ultraviolet irradiation; step 3, collecting intramuscular fat cells or subcutaneous fat cells after cross-linking treatment, and performing lysis under the action of a cell lysis solution to obtain a lysis solution; step 4, adding magnetic beads into the lysate, and carrying out repeated resuspension and elution to obtain an eluent; and 5, separating the RNA binding protein from the magnetic beads, and incubating. According to the method, intramuscular fat cells or subcutaneous fat cells are treated by adopting a UV cross-linking solidification method, and then RNA binding protein is captured through magnetic beads, so that the RNA binding protein is successfully obtained.

The invention relates to the technical field of vaccines, and discloses a preparation method and application of an anti-tumor vaccine based on an autoantigen and a synthetic antigen. The anti-tumor vaccine comprises a tumor autoantigen and a synthetic antigen, wherein the tumor autoantigen is extracted from a tumor cell membrane, a tumor cell lysate, a tumor cell secretion, an exosome antigen or an antigen released by a tumor microenvironment; the synthetic antigen is selected from at least one of a tumor associated antigen, a tumor specific antigen, a personalized neoantigen, a pathogen antigen, a biotoxin, a biomolecule antigen and coding nucleic acid of a peptide type antigen in the antigens; the coding nucleic acid is DNA or mRNA. The vaccine molecule composed of the tumor autoantigen and the synthetic antigen is easy to prepare, has a strong anti-tumor immune effect and good biological safety, solves the problems that the traditional tumor autoantigen has no specific precise target spot and the synthetic antigen is single in immunogen, and has a wide application prospect.

The invention relates to the technical field of protease separation, in particular to a novel earthworm protease separation and purification process. According to the present invention, the ultra-high speed separation, the molecular weight spectrum comparison extraction and the affinity chromatography purification scheme are adopted, such that the different biological macromolecules in the earthworm can be effectively separated, the earthworm protease-containing cell lysis solution can be concentrated, and most of the impurities can be removed so as to obtain the high-purity protease solution, the method is simple in operation and does not depend on complex chemical reactions, contact between protein and chemical reagents can be reduced, meanwhile, damage of high temperature to earthworm protease is avoided through a low-temperature centrifugation scheme, the enzymatic activity of earthworm protease is kept, high-purity and high-activity earthworm protease is obtained through affinity chromatography purification, operation is easy, and the method is suitable for industrial production.

The present application provides an activity analysis detection method for a stress-induced mesenchymal stem cell extract, comprising step one, culturing human primary articular chondrocytes; step two, aspirating the culture fluid, adding an equal volume of a composition to be tested, and incubating, wherein the composition to be tested includes a mesenchymal stem cell extract; step three, then adding an equal volume of a lipopolysaccharide solution with a concentration of 80ng / mL-120ng / mL, continuing to incubate, and collecting a cell lysate, which is used to detect inflammatory cytokines. The present application determines the optimal use concentration and cell incubation time of a mesenchymal stem cell extract (denoted as Alituo) and a lyophilized composition (denoted as Alip) and a lipopolysaccharide solution by measuring cell viability and the levels of inflammatory factors IL-6, IL-1β, and TNF-α. By detecting cell viability and inflammatory factors, the quality of the mesenchymal stem cell extract can be efficiently evaluated, and the activity analysis detection method has high repeatability and good stability.

The application belongs to the technical field of microbial fermentation, and particularly relates to a kind of cytolysis product of mixed bacteria fermentation and a preparation method. The cytolysis product is prepared by mixed bacteria fermentation of two kinds of aerobic probiotics, i.e. Lactobacillus plantarum and Aspergillus oryzae are used as fermentation strains to carry out mixed bacteria fermentation treatment on rice milk, and membrane filtration and column chromatography system are used for filtration and enrichment, so as to obtain a cytolysis fermentation product filtrate rich in kojic acid, the content of kojic acid is greater than 200 g / L, which is significantly improved compared with conventional process. The obtained cytolysis product has no bad smell and clear color, so that the beneficial function of kojic acid can be played only with a low addition amount in cosmetics, and the addition of odor masking and color correcting auxiliaries is reduced, and the cytolysis product can be used for preparing cosmetics with skin repair, whitening and other functions.

The invention relates to the technical field of biological analysis and detection, and particularly discloses a streptococcus mutans detection kit based on an anchored CRISPR-Cas12a enhanced paper-based micro-fluidic chip and a detection method. The detection kit comprises an RAA solution or RAA freeze-dried powder, magnesium acetate or a magnesium acetate solution, TMB or a TMB solution, an RAA forward primer, an RAA reverse primer, a cell lysis buffer, a buffer system and a paper-based micro-fluidic chip. The detection method comprises the following steps: establishing a standard curve and detecting a sample. The detection kit is used for detecting the content of streptococcus mutans and has the characteristics of high efficiency, rapidness, specificity, sensitivity, portability and the like, and the method is simple and convenient to operate, low in detection cost and capable of being applied to on-site detection of streptococcus mutans.

The invention discloses a cell-free cloning reaction system, a kit and application of the cell-free cloning reaction system. The system comprises a cell lysis solution, an inserted DNA fragment and a linear vector, the cell lysis solution is obtained from Escherichia coli which is subjected to induced expression and then subjected to lysis, and a lambda Red system and a ligase system are expressed in the Escherichia coli; the lambda Red system comprises Exo protein, Beta protein and Gam protein; and the ligase system comprises DNA ligase. The optimized lambda Red system and the efficiently expressed DNA ligase system can complete efficient splicing of multi-fragment DNA within 15 minutes at 37 DEG C, still maintain a high positive rate in a multi-fragment assembly system, show an effect superior to Gibson cloning, and expand the applicability of the Gibson clone in complex pathway synthesis and gene line reconstruction.

Disclosed are systems and methods for producing renewable alcohols by fermentative processes using cell lysate or a combination of cell lysate and whole cells, as well as the cell lysate itself. The disclosure also provides processes and systems for converting renewable alcohols to transportation fuels, such as, gasoline, jet fuel, and diesel fuels.

The present invention discloses a method for producing perillaldehyde and / or perillyl lactone, comprising: contacting a first component with a second component to produce perillaldehyde and / or perillyl lactone; wherein the first component is selected from components containing lysine-2-diol, and the second component is selected from enzymes or microorganisms or their cell lysates or enzyme extracts capable of converting lysine-2-diol into perillaldehyde and / or perillyl lactone.

The present invention belongs to the field of extraction of microbial phospholipids, and specifically discloses a method for efficiently purifying phospholipids from coenzyme Q10 fermentation powder, which comprises the following steps: 1) mother liquor culture, 2) seed tank culture, 3) fermenter culture, 4) centrifugation of fermentation broth, 5) high-pressure homogenization, 6) centrifugation of cell lysate, 7) vacuum freeze-drying, 8) subcritical extraction, 9) acetone extraction, 10) rotary evaporation; by adjusting the medium components and optimizing the culture process, the present invention can achieve high-density culture of Agrobacterium tumefaciens cells, obtain a large amount of cells, in the fermentation broth, the coenzyme Q10 content can reach 5.1 g / L of fermentation broth, and the phospholipid content can reach 13.5 g / L of fermentation broth. Moreover, the present method also discloses a separation and purification method for efficiently extracting high-purity phospholipids from coenzyme Q10 fermentation powder, turning waste into treasure, with good economic benefits, and the steps are simple and mild, with low requirements for equipment, and are environmentally friendly.

The present invention discloses a ratio-type dopamine fluorescent probe and its application for the kinetic highly selective determination of dopamine. The method for determining dopamine is as follows: After adjusting the pH value of a 3,5-dihydroxybenzoic acid solution with a base, it is used as a fluorescent probe. After reacting with the test solution, the fluorescence emission spectra are measured respectively at excitation wavelengths of 340 ± 10 nm and 375 ± 10 nm. The dopamine concentration is calculated through the ratio equation of the intensity ratio between the strongest peak of the reaction product of 3,5-dihydroxybenzoic acid and dopamine and the strongest peak of 3,5-dihydroxybenzoic acid in the fluorescence emission spectrum. The 3,5-dihydroxybenzoic acid probe prepared by the present invention has excellent selectivity, very high stability, high sensitivity based on the ratio-type probe, extremely low detection limit and good biocompatibility, and can be used for the detection of dopamine. At present, it has been successfully applied to the selective detection of dopamine in human urine samples and PC12 cell lysates.

The invention discloses an efficient ribosome imprinting sequencing library construction method, and belongs to the technical field of biology. In the enzyme digestion treatment step, a mixed enzyme system of RNase I and micrococcus nuclease is adopted to treat the cell lysis buffer, and the trinucleotide periodicity of RPFs can be remarkably improved to 80% or above. In the target fragment recovery step, the sucrose gradient ultracentrifugation step is omitted, the RPFs are directly recovered by gel electrophoresis, a plurality of enzyme reaction steps in library construction are optimized and integrated, and streptavidin magnetic beads are used for solid phase purification, so that the whole library construction period is greatly shortened from 7 days to 2.5 days. The method does not depend on special equipment such as ultracentrifugation, and can be completed in a conventional molecular biology laboratory. Besides, a multi-enzyme synergistic RNA terminal one-step modification system is innovatively constructed, single-tube one-step completion is realized, the terminal treatment efficiency and consistency are improved, and a key guarantee is provided for obtaining a high-quality and high-complexity Ribo-seq library.

The invention relates to a gene delivery system for cytoplasm specific expression of PPM1K as well as a preparation method and application of the gene delivery system, and belongs to the technical field of preparation of gene delivery systems. The gene delivery system is preferably an adeno-associated virus (AAV), and the preparation method of the gene delivery system specifically comprises the following steps: inserting a PPM1K gene or a functional variant thereof into a vector, and adding a detectable label at a C terminal to form a recombinant plasmid; transforming the recombinant plasmids into competent escherichia coli, amplifying, and screening and extracting plasmids to obtain recombinant AAV plasmids; co-transfecting the recombinant AAV plasmid, the helper plasmid and the capsid plasmid into packaging cells by using a transfection reagent, collecting the cells and culturing supernate after transfection, and adding a freeze thawing buffer solution for repeated freeze thawing to obtain a cell lysis solution; centrifuging the cell lysis solution, taking supernate, purifying and filtering. According to the invention, the specific expression of the cell compartment of the therapeutic gene is realized through the vector design, and the obvious therapeutic effect and the safety advantage are shown.

The present invention relates to a method for recovering high-purity black mass from a used lithium ion secondary battery and, more particularly, to a method for recovering high-purity black mass through: an alkaline thermal hydrolysis step of stirring a cell lysate of the used lithium ion secondary battery in an alkaline aqueous solution to dissolve and separate impurities contained in the cell lysate into an aqueous solution; and a low-temperature pyrolysis step of decomposing and removing organic compound components, such as an organic binder.

The utility model relates to the technical field of cell lysis solution separation, and discloses a cell lysis solution separation device which comprises a separation box, a material conveying pump fixedly mounted at the upper part of the separation box, a recovery pipe communicated with the output end of the material conveying pump, a solution inlet pipe communicated with one end of the recovery pipe, and a treatment box communicated with the left side of the separation box, the connecting assembly is arranged on one side of the separation box and comprises a filtering mechanism arranged on the left side of the separation box, a separation mechanism is arranged in the separation box, the input end of the material conveying pump communicates with the bottom of the separation box through a connecting pipe, and a liquid outlet pipe communicates with the left side of the treatment box; a valve is arranged at the outer end of the liquid outlet pipe, and one end of the first spring is fixedly connected with the sliding block. The device disclosed by the utility model solves the problem that the separation effect of the subsequent cell lysis solution is influenced due to the fact that filter residues possibly fall into a separation box when a filter membrane is disassembled by an existing device.

The invention discloses a protein post-translational modification detector based on a GST fusion histone H3 fragment mixture as well as a preparation method and application of the protein post-translational modification detector. The protein post-translational modification detector comprises 9 segmented fragments H3F1-H3F9 of histone H3. The prepared GST is fused with an H3 fragment (H3F1-H3F9) mixed plasmid; the mixed plasmid is transformed into BL21-DE escherichia coli; GST fusion H3 fragment mixture expression is carried out; purification is carried out; after the mixture is combined with GSH resin, incubation with a cell lysis solution is carried out for PTM; the GST label of the mixture after PTM is cut off by using precession TM protease, and the H3F1-H3F9 mixture is released into a solution; if a signal peak of the corresponding molecular weight is detected in the mass spectrum, PTM of the organic acid can be confirmed.

The present application belongs to the field of biomedical technology, and particularly relates to a cell-derived extracellular matrix and a preparation method and application of a cell culture plate coated with the cell-derived extracellular matrix. The present application cultures human astrocytes in a cell culture plate, and makes the human astrocytes secrete extracellular matrix. After the human astrocytes are added into a cell lysate for cell removal treatment, nucleic acid fragmentation treatment is performed to obtain the cell-derived extracellular matrix. The present application has relatively simple operation, short time period, and can realize batch production of extracellular matrix without external virus carrying and stable components. The obtained cell-derived extracellular matrix is resistant to storage, has good stability, is highly similar to a natural brain environment when coated on a cell culture plate, has good biocompatibility with cells, can be applied to tissue repair and regeneration as an ideal biomedical material, can regulate the progress and efficiency of human dermal fibroblast HDF transdifferentiation into neurons, and can obtain in-vitro survival neurons.

Systems and methods for assessing mRNA in vivo and / or in vitro stability are disclosed. Some embodiments methods obtain RNA indexed or barcoded RNA molecules which are then tested against various conditions including stability inside of cells, stability in cell lysate, and stability in solution (e.g., for storage and / or transportation). Additional embodiments describe methods to determine degradation points with single base resolution.

The present invention provides engineered acid alpha-glucosidase (GAA) polypeptides and compositions thereof. In some embodiments, the engineered GAA polypeptides have been optimized to provide increased expression, stability at neutral pH, and activity in cell lysates. The invention also provides methods for utilization of the compositions comprising the engineered GAA polypeptides for therapeutic and other purposes.

The invention discloses a metagen which is prepared from bifidobacterium longum CCFM1508 and has the effects of regulating skin ceramide de novo synthesis and improving skin health, and belongs to the technical field of microorganisms and medicines. The bifidobacterium longum CCFM1508 and the metagen prepared from the bifidobacterium longum CCFM1508, provided by the invention, have the effects of promoting ceramide synthesis and improving skin health by external use and oral administration, and are specifically shown as follows: a thallus lysate and a fermented supernatant of the bifidobacterium longum CCFM1508 can promote HaCaT cells to secrete ceramide in the aspect of external use, and the thallus lysate can remarkably improve the activity of the HaCaT cells. And the viable bacteria and the postbiotics of the bifidobacterium longum CCFM1508 which are orally taken have obvious effects in skin apparent indexes, the content of cerayl and proinflammatory factors. Therefore, the bifidobacterium longum has huge application prospects in preparation of external or oral medicines for promoting synthesis of skin ceramide and improving skin health and preparation of food, health care products or cosmetics.

The present invention relates to a method for virus capture or separation. More closely, the invention relates to a method for direct influenza and adenovirus capture using magnetic beads. The method allows direct separation from crude cell lysate in a rapid manner.

The present invention relates to the field of microorganisms, and specifically, to a Lactiplantibacillus plantarum YW99, its cell lysate, and their antibacterial applications. Lactiplantibacillus plantarum ( Lactiplantibacillus plantarum ) YW99 has been deposited in the General Microbiological Center of the China General Microbiological Culture Collection Center, with the deposit number of CGMCC No. 33160. The cell lysate of Lactiplantibacillus plantarum YW99 is prepared by a series of operations after the YW99 strain is heat-inactivated. It has good safety, will not cause toxic and side effects to the human body, has a certain function of improving the body's immunity, and can show different degrees of antibacterial ability against Candida albicans, Gardnerella, Pseudomonas aeruginosa, Aspergillus brasiliensis and other strains by changing the concentration of the YW99 cell lysate. It has good application prospects in female health products, hygiene products, daily chemical products, and disinfection products.

A method for rapid biocementation of a material under low moisture conditions includes mixing a cell lysate of a urease-producing microbe with the material and, optionally, a calcium source and / or a carbon source, under initial moisture conditions of less than about 15% by weight to form a cementing composition; incubating the cementing composition for a selected time under pressure conditions of between 0 to 1500 bars to form an incubated mixture; and curing the incubated mixture for hours to weeks.

The invention discloses an electrochemiluminescence detection method of PARP enzyme, a kit and application thereof. The electrochemical luminescence detection method for detecting PARP enzyme comprises the following steps: (1) mixing human whole blood, a PARP inhibitor and a PARG inhibitor, incubating, and centrifuging to obtain human peripheral blood mononuclear cells; (2) collecting a human peripheral blood mononuclear cell precipitate for cracking, detecting the total protein concentration, adding a sample, adding a cell lysis solution into an electrochemical luminescent plate for incubation, and adding a PARP enzyme detection antibody for re-incubation; and (3) adding a detection secondary antibody for incubation to form a capture antibody-PARP enzyme-detection antibody-secondary antibody complex, and adding plate reading liquid for plate reading. The electrochemical luminescence detection method and the kit have the characteristics of high sensitivity, high specificity, high accuracy, high precision and simple operation method, and can quantitatively detect the content of PARP enzyme.

The present application relates to an assay method using affinity crosslinking of DNA-linked ligands to proteins to enable small molecule screening and determination of apparent affinity constants of ligands to the proteins. This method has been applied to determine 96 compounds' dissociation constants to a protein target simultaneously, and directly determine a compound's IC50 against five protein targets concurrently in crude cell lysates. Additionally, this approach was used to screen a Library of Pharmacologically Active Compounds (LOPAC) library against dihydrofolate reductase (eDHFR), enabling the discovery of a novel eDHFR inhibitor (IC50=7.9 μM). An assay kit and the method of uses are within the scope of this disclosure.

The present invention provides a Bacillus strain, a biocontrol microbial agent prepared from the bacillus strain, and a use thereof. The Bacillus strain provided in the present invention is Bacillus subtilis, named QY2, for which the deposition institution is the China General Microbiological Culture Collection Center, the deposition address is the Institute of Microbiology, Chinese Academy of Sciences, No. 1 Beichen West Road, Chaoyang District, Beijing, the deposition center number is CGMCC No. 31373, and the deposition time is 19 July 2024. The Bacillus stain QY2 of the present invention, whether in the form of viable bacteria, a supernatant, or a cell lysate, exhibits inhibitory effects on various pathogens. The Bacillus stain QY2 has strong viability, wide applicability in terms of nutrient sources, temperature, humidity, and pH, no pathogenicity to fruits and vegetables, and relatively good tolerance to broad-spectrum microbicides.

The present invention relates to the provision of trifunctional crosslinker compounds comprising (i) a moiety reactive with an amino acid side chain for selective labelling of proteins, (ii) a cleavable linker moiety and (iii) an affinity moiety for the detection, isolation and purification of captured proteins. The invention further relates to uses of the cleavable crosslinking reagents in methods for detecting, labelling, identifying and characterizing proteins and protein interactions in or on the surface of intact cells, cell lysates and / or protein mixtures.