100 results about "Cell lysates" patented technology

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 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 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 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.

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.

The invention provides a dendritic cell vaccine for treating bone and soft tissue sarcoma as well as a preparation method and application of the dendritic cell vaccine. The dendritic cell vaccine comprises mature dendritic cells obtained by co-culturing sarcoma cell lysate and immature dendritic cells. The lysates of seven sarcoma cells are used as dendritic cell vaccine antigens, the problem that an existing DC vaccine is single in loaded antigen is solved, antigen presentation and T cell activation are enhanced, wide immune response is excited, immune escape is reduced, and the dendritic cell vaccine is not limited by the type of MHC of a patient, is high in universality and can benefit more patients.

The present application relates to a bacillus, a biocontrol agent made from the bacillus and application, and belongs to the technical field of biological bacterial agents. The bacillus provided by the present application is Bacillus subtilis, named ZZ1, and the preservation unit is the General Microbiological Center of China Microorganism Bacterial Strain Preservation Management Committee, the preservation address is the Institute of Microbiology of Chinese Academy of Sciences, No. 3, Xili, Beichen, Chaoyang District, Beijing, the preservation center number is CGMCC No. 31374, and the preservation time is July 19, 2024. The bacillus ZZ1 has certain inhibition effect on different pathogenic bacteria, whether it is a living bacterium, a supernatant or a cell lysate. The bacillus ZZ1 has strong living ability, and is suitable for a wide range of nutrition sources, temperature and humidity, and pH. The bacillus ZZ1 has no pathogenicity to fruits and vegetables, and has good tolerance to broad-spectrum bactericides, and has great application prospect.

This invention discloses a method for protein extraction and mass spectrometry pretreatment from methanogenic bacteria, belonging to the field of protein extraction technology. The method involves pretreating a methanogenic bacteria enrichment culture to remove inorganic salt ions and other impurities, resulting in a pretreated culture. Then, a lysis buffer composed of sodium dodecyl sulfate, Tris-HCl buffer, and a protease inhibitor is added to the pretreated culture, followed by ultrasonic disruption to obtain a cell lysate, from which a crude protein extract solution is obtained. Finally, the protein extract is extracted from the crude protein extract solution. This invention significantly improves protein extraction efficiency and peptide purity by optimizing the extraction process and subsequent enzymatic digestion, while preserving the methanogenic activity of the extracted protein as much as possible, providing more accurate and reliable sample support for downstream protein mass spectrometry analysis.

The invention provides an all-round nuclease mutant as well as a preparation method and application thereof, and relates to the technical field of biology. Compared with a wild type, the totipotent nuclease mutant provided by the invention has higher enzyme activity and temperature resistance and better stability, can be applied to industrial applications such as vaccine purification, biological pharmacy residual DNA removal, cell lysis solution treatment and the like, and has obvious technical advantages and popularization value. The preparation method of the totipotent nuclease mutant provided by the invention is simple and convenient, the expression quantity of the totipotent nuclease mutant is effectively improved by further optimizing host cells, an expression vector, an inclusion body inducing expression method and a renaturation mode, and the problem that the activity of the totipotent nuclease is obviously reduced after renaturation is relieved.

This invention discloses an umbilical cord mesenchymal stem cell membrane for treating hair loss, its preparation method, and its application, belonging to the field of biomedical technology. The preparation method of the umbilical cord mesenchymal stem cell membrane includes the following steps: obtaining human umbilical cord mesenchymal stem cells, digesting, centrifuging, washing, and resuspending them to obtain a resuspended cell solution; subjecting the resuspended cell solution to repeated freeze-thaw cycles to break the cells, obtaining a cell lysate; and subjecting the cell lysate to differential centrifugation to obtain the umbilical cord mesenchymal stem cell membrane. This umbilical cord mesenchymal stem cell membrane retains the natural cell membrane structure and functional proteins, has a negatively charged surface, and exhibits good dispersibility and stability. It can improve the survival rate of dermal papilla cells without introducing live cells and effectively regulate the skin microecology and metabolite balance, demonstrating high safety, good biocompatibility, and excellent hair growth promotion effects.

The present application relates to a bacillus, a biocontrol agent made from the bacillus and an application thereof, and belongs to the technical field of biological bacterial agents. The bacillus provided by the present application is Bacillus subtilis, named QY2, and the preservation unit is the General Microbiological Center of China Microorganism Bacterial Strain Preservation Management Committee, the preservation address is the Institute of Microbiology of Chinese Academy of Sciences, No. 3, Xili, Beichen, Chaoyang District, Beijing, the preservation center number is CGMCC No. 31373, and the preservation time is July 19, 2024. The bacillus QY2 has certain inhibition effects on different pathogenic bacteria, whether it is a living bacterium, a supernatant or a cell lysate. The bacillus QY2 has strong living ability, and is suitable for a wide range of nutrition sources, temperature and humidity and pH. The bacillus QY2 has no pathogenicity to fruits and vegetables, and has good tolerance to a wide range of fungicides, and has great application prospect.

Methods and systems are provided for the purification of native and recombinant Ribulose- 1,5- bisphosphate Carboxylase / Oxygenase (RuBisCO) from cellular lysates using liquid-liquid phase separation (LLPS). The invention enables scalable and cost-effective extraction of highly pure RuBisCO complexes from plant and bacterial sources while maintaining functional protein integrity. The methods and compositions disclosed herein address key obstacles to RuBisCO isolation and offer commercial advantages for protein production and utility in nutritional formulations.

The present application relates to a kind of cytoplasm-specific expression PPM1K gene delivery system and its preparation method and application, belong to gene delivery system preparation technical field.The gene delivery system is preferably adeno-associated virus (AAV), and its preparation method specifically includes the following steps: PPM1K gene or its functional variant is inserted into vector, detectable label is added at C-terminal, and recombination plasmid is formed;Recombinant plasmid is transformed into competent e. coli and is amplified, after plasmid is screened, recombination AAV plasmid is obtained;Recombinant AAV plasmid, auxiliary plasmid and capsid plasmid are co-transfected into packaging cell using transfection reagent, after transfection, cell and culture supernatant are collected, freeze-thaw buffer is added repeatedly freeze-thaw, and cell lysate is obtained;After centrifugation of cell lysate, supernatant is taken, and purified filtration is carried out.The present application realizes the cell compartment specificity expression of treatment gene by vector design, and shows significant treatment effect and safety advantage.

Provided herein are methods of producing a virus from a culture of a host cell. The method comprises providing a culture of a host cell that has been infected with the virus; contacting the host cell with a first buffer comprising a detergent and incubating the host cell in the presence of the first buffer for a first period of time, thereby producing a cell lysate; contacting the cell lysate with a second buffer comprising an endonuclease for a second period of time to degrade host cell nucleic acids; and collecting the virus.