54 results about "Cell lysates" patented technology

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.

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 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 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 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 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.

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.

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.

The invention discloses an alkynylation dihydroartemisinic acid probe, a biotin-dihydroartemisinic acid probe and a preparation method and application of the alkynylation dihydroartemisinic acid probe. The structural formula of the alkynylated dihydroarteannuic acid is as shown in formula 1. And a'biotin-dihydroartemisinic acid 'probe structure is constructed. And co-incubating the probe and a cell lysis solution, co-incubating a lysis solution-probe mixed solution and streptavidin magnetic beads, and eluting to obtain the drug-specific binding protein. According to the invention, the action target of dihydroartemisinin in disease treatment can be quickly, simply and conveniently found.

The invention belongs to the field of membrane separation technology and bioengineering, and particularly relates to an anti-pollution composite nanofiber membrane for high-precision and high-flux separation of protein and a preparation method of the anti-pollution composite nanofiber membrane. According to the separation membrane, a hydrophilic polymer and zwitterionic polymer blended nanofiber membrane prepared through electrostatic spinning serves as a base material, zwitterionic polymers are grafted to the surfaces of fibers through the surface-initiated atom transfer radical polymerization (ATRP) technology, and a separation functional layer with excellent anti-pollution performance is formed. According to the invention, non-specific adsorption of protein and membrane pollution are effectively inhibited by constructing a high-density hydration layer, and the membrane has high porosity and high connectivity of a nanofiber-based membrane and super-hydrophilic and anti-pollution characteristics of a zwitterionic layer at the same time. The composite membrane has high selectivity, high rejection rate and excellent cycling stability on target protein in complex biological samples (such as serum, cell lysis buffer and fermentation liquor), and solves the problems that a traditional polymer membrane is easy to pollute, low in efficiency and poor in precision in protein separation.

The present disclosure provides a method of purifying polysaccharides from a cell lysate, comprising partially purifying the cell lysate comprising an impurity and a polysaccharide to obtain a clarified crude lysate; mixing the clarified crude lysate with a neutralization solution comprising a salt to form a neutralized lysate; mixing the neutralized lysate with a precipitation solution comprising cetyltrimethylammonium bromide to form a first supernatant and a first precipitate; and separating the first precipitate from the first supernatant, wherein the polysaccharide is located in the first supernatant. The present disclosure further provides a method of making a polysaccharide vaccine. Also provided are vaccines, delivery systems, compositions and polysaccharides made by the methods described herein.

This invention discloses a method for isolating and extracting PDRN from microorganisms. The method includes: collecting bacterial cells from a microbial culture medium; washing and resuspending the bacterial cells to obtain a bacterial resuspension; disrupting the bacterial resuspension and centrifuging it to collect the supernatant, obtaining a bacterial cell lysate; adding a soluble salt and an organic solvent to the bacterial cell lysate, mixing thoroughly, allowing it to stand or centrifuging to separate the phases, forming a three-phase salting-out extraction system consisting of an upper phase, an intermediate phase, and a lower phase; collecting the lower phase; purifying and drying the collected lower phase to obtain PDRN powder. This invention uses microbial cells as raw material and obtains high-purity PDRN in one step through salting-out extraction. The obtained PDRN has a polydispersity index (PDI) of 1.1–1.3, with uniform DNA fragment length and a narrow molecular weight distribution. The method of this invention has the advantages of simple process, low cost, uniform product fragments, narrow molecular weight distribution, and high purity, making it suitable for large-scale industrial production.

The present invention relates to a cell lysate wherein the lysis buffer comprises magnesium acetate, the phosphorylation inhibitor K3L, the agonist of dephosphorylation GADD34 and a caspase inhibitor for use in, or as, a cell-free system and a method for preparing a cell-free protein synthesis system. Further provided is a method for expressing a protein in the cell-free protein synthesis system, a method for screening for a candidate compound for an effect on a target molecule and a use of the cell-free protein synthesis system. The present invention also provides a kit for the above-mentioned cell-free systems.

The application discloses a cell lysate of hAMCs cultured in vitro and a determination method of neurotrophic factor content of culture supernatant, wherein the hAMCs are cultured in vitro, when the cells grow to 80%-90% confluence, subculture is carried out, until the cells reach 80% confluence, then centrifugation is carried out to obtain the culture supernatant of hAMCs, PBS buffer is added to the hAMCs from which the supernatant is removed, centrifugal cleaning is carried out, the cells are broken in an ultrasonic cell disrupter to obtain the cell lysate of hAMCs, and finally, the neurotrophic factor content of the culture supernatant and the cell lysate of hAMCs is determined by using an ELISA method.

The invention provides an acellular matrix for treating stress urinary incontinence as well as a preparation method and application of the acellular matrix. Splitting the soft tissue by using a cell lysis solution to remove cell debris; removing nucleic acid, degreasing and then micronizing; and adding the soft tissue particles and the soft tissue cross-linked particles into a phosphate buffer solution according to a ratio, mixing and emulsifying to obtain the acellular matrix for injection for treating stress urinary incontinence. The cell removal rate reaches 99%, the fat residue is less than 0.3%, and the injection is injected around the sphincter of the urethra (middle section or bladder neck) and / or in the urethra according to 0.5 ml / point and 3-4 points during clinical application to play roles in filling, connecting and repairing tissues, so that the damaged urethra supporting structure is repaired, the structural integrity and strength of connective tissues and the sphincter around the urethra are maintained, and the application prospect is wide. Therefore, the symptom of stress urinary incontinence is improved.

The application provides a method for separating and purifying cell surface proteins and application thereof. The method comprises the following steps: culturing cells to be separated, adding an azide solution to the cells for incubation, then adding a Tris-HCl buffer to terminate the reaction, so as to label the cell surface proteins with azides; then adding a TNTE buffer and a protease inhibitor to lyse the cells, centrifuging to obtain cell lysates labeled with azides; incubating the cell lysates labeled with azides with phosphine-biotin-streptavidin magnetic beads, so as to enrich the cell surface proteins labeled with azides, and obtain magnetic beads containing the cell surface proteins. The method can effectively separate the cell surface proteins, has low background, high efficiency and good repeatability, and can be applied to quantitative analysis of cell surface proteomes.

The present application belongs to the field of fermentation engineering and biomass resource utilization, and particularly relates to preparation of a composite nanofiber membrane by utilizing fermentation bacteria after fermentation production is completed. In the present application, the fermentation bacteria are subjected to chemical lysis to release biological macromolecules. The lysis solution of the bacteria after lysis contains biological macromolecules such as proteins, nucleic acids, peptidoglycan and flocculating agent chitosan. Then, the bacteria lysis solution is blended with starch and PEO, and a composite nanofiber is prepared by electrospinning, so as to realize productized application of the waste fermentation bacteria.

The invention discloses an application of a mitochondria-rich cell disintegration body in treatment of myocardial ischemia reperfusion injury. The invention provides an application of a mitochondrial-rich cell disintegration body in preparation of a product for preventing and / or treating myocardial ischemia-reperfusion injury. The cell disintegration body is extracted from mitochondrial donor cells treated by cytochalasin D. The invention further provides a preparation method of the mitochondrial-rich cell disintegration body. The mitochondrial-rich cell disintegration body has ultrahigh mitochondrial loading capacity and functional activity and excellent in-vivo and in-vitro stability; the functional mitochondria can be efficiently delivered to a heart injury area, the myocardial protection effect is achieved by activating an endothelial cell autophagy pathway, the heart function is remarkably improved, the infarct area is reduced, and the good clinical application prospect is achieved.

The present application relates to a bacillus velezensis, a biocontrol agent prepared from the bacillus velezensis and an application thereof, and belongs to the technical field of biological bacterial agents. The bacillus velezensis provided by the present application is bacillus velezensis B. velezensis, named JK1, the preservation unit is China General Microbiological Culture Collection Center, the preservation address is Institute of Microbiology, Chinese Academy of Sciences, No. 1, Beichen West Road, Chaoyang District, Beijing, the preservation center number is CGMCC No. 31375, and the preservation time is July 19, 2024. The bacillus JK1 has excellent inhibition effect on different pathogenic bacteria whether it is a living bacterium, a supernatant or a cell lysate. The bacillus JK1 has strong living ability, and is suitable for a wide range of nutrition sources, temperature and humidity and pH. The bacillus JK1 has no pathogenicity to fruits and vegetables, and has good tolerance to a broad-spectrum bactericide, and has great application prospect.

Embodiments of the disclosure include treatments of subconcussive and / or concussive brain damage by administering fibroblasts and / or fibroblasts cultured with one or more types of immunocytes. In one specific embodiment fibroblasts are cultured with monocytes in the presence of patient-specific T cells, and subsequently the T cells are re-administered into the patient. In one particular embodiment, products derived from fibroblast-immunocyte mixtures are comprised of cellular lysate, apoptotic bodies, exosomes, and / or other microvesicles. In one embodiment, the fibroblast cells and / or products derived from the fibroblast cells are administered subsequent to one or multiple head injuries. In other embodiments, products are administered in combination with neurorestorative and / or neuroprotective interventions.

The present invention relates to a method for characterizing antibiotic resistance of a microorganism comprising the steps of: (a) providing a reference mass spectrum of an antimicrobial compound, an enzyme modification product thereof, a molecular target thereof or a substrate compound of a modifying enzyme thereof; (b) exposing a microorganism, its cell lysate, its growth medium supernatant to the antimicrobial compound or the substrate compound in aqueous liquid, thereby providing an exposed sample; (c) acquiring a mass spectrum of the exposed sample; (d) comparing the mass spectrum obtained in step c) with the reference mass spectrum obtained in step a), and (e) determining from said comparison whether a modification of the antimicrobial compound, its modification product, its molecular target or the substrate has occurred after said exposure, and establishing that the microorganism is potentially resistant to the antimicrobial compound when the modification is observed.