434 results about "Cell free" patented technology

This invention provides an in vitro cell-free protein synthesis system, its reagent kit, and its applications, belonging to the field of protein synthesis technology. This invention improves an in vitro cell-free protein synthesis system based on Kluyveromyces lactis by adding exogenous L-arabinose, significantly enhancing the system's protein synthesis capacity. Furthermore, it provides a more efficient and higher-throughput in vitro protein synthesis reagent kit and a method for synthesizing exogenous proteins. The improvements provided by this invention can be achieved without molecular modification, are simple and convenient to operate, and save costs.

The invention provides an ELISA (Enzyme Linked Immunosorbent Assay) kit for detecting an anti-MDA5 antibody and a preparation method of the ELISA kit. According to the kit, full-length human MDA5 protein (1025 amino acids) expressed by a cell-free protein expression system under the low-temperature condition of 22 DEG C is adopted as a coating antigen, and a disulfide bond enhancer E6820 is added into the system to promote correct folding of the protein. The kit comprises a microwell plate coated with a full-length MDA5 antigen, an enzyme-labeled anti-human IgG secondary antibody, a standard substance with the concentration range of 1-243 ng / mL, a chromogenic substrate and an optimized confining liquid system. The invention solves the problem of insufficient sensitivity caused by the use of fragment antigens in the prior art, and can be used for early diagnosis and illness monitoring of dermatomyositis-related interstitial lung diseases.

The application provides an in-vitro cell-free protein synthesis system and kit containing exogenous magnesium ions and application thereof, including a D2P system (DNA-to-Protein system) and an mR2P system (mRNA-to-Protein system), and belongs to the technical field of protein synthesis. The in-vitro cell-free protein synthesis system containing exogenous magnesium ions adopts magnesium aspartate as a novel magnesium ion source, and especially in a eukaryotic cell-free system, compared with a traditional magnesium ion source, can significantly improve the protein synthesis efficiency and protein expression amount. A more efficient and higher-throughput in-vitro protein synthesis kit and a synthesis method of exogenous proteins are also provided, and the kit has the advantages of simplicity, convenience and low cost.

An acellular product may be derived from human placenta and may be used in various scenarios for wound healing. Because the product may be acellular, the product may be processed for storage and transportation with minimal degradation. The product may include various scaffolding such as biomaterials or human tissue, and the scaffolding may be infused with various plasmas and agents. The cell-free treatment may maintain the biological activity of many therapeutic agents found within cells and may possess multiple structural components to support cellular attachment. The structural components or scaffolds may function as a reservoir of highly diffusible chemotactic and cellular-programming factors that may be useful to treat injury and disease.

A method for improving degradation efficiency of corn straw at low temperature comprises the steps that cell-free culture fluid PfCFs prepared from Pseudomonas fragi D12 participates in fermentation and degradation of the corn straw, the classification name of the strain D12 is Pseudomonas fragi D12, the strain D12 is preserved in China Center for Type Culture Collection, the preservation address is No. 299, 8th road, Wuchang District, Wuhan City, Hubei Province, China, and the preservation number is CGMCC NO. The preservation number is CCTCC (China Center For Type Culture Collection) NO: M2025126, and the preservation date is January 14, 2025. The acellular culture solution of the pseudomonas rubrassicae D12 can promote the growth of various normal-temperature bacteria and fungi functional bacteria at low temperature and improve the cold resistance of the functional bacteria, and CFS of the pseudomonas rubrassicae D12 also contains related enzymes capable of degrading lignin; at a low temperature, the PfCFs synchronously promotes the co-growth of bacteria bacillus subtilis and fungi Irpex lacteus, so that the degradation efficiency of the functional bacteria on lignocellulose in the corn straws is improved.

96 New PCT-Patent Application based on EP 24 186 635.9 Insempra GmbH Vossius Ref.: AG4141 PCT S3 Abstract The present invention relates to a linear double-stranded deoxyribonucleic acid (dsDNA) molecule comprising one or more Tus protein (Tus) binding site(s) at the 5'-terminus, one or more Lac repressor protein (LacI) binding site(s) at the 3'-terminus, and a segment comprising a DNA sequence of interest (DOI) between said Tus binding site(s) and said LacI binding site(s). The present invention further relates to a non-naturally or naturally occurring RNA molecule as encoded by or transcribable / transcribed from the linear dsDNA molecule of the invention, and to an expression DNA cassette comprising a promoter, an RBS, a GOI encoding a POI, and one or more LacI binding site(s), wherein said expression cassette does not comprise a terminator sequence between the 3´ end of said GOI and said one or more LacI binding site(s). The present invention also relates to a non-naturally or naturally occurring RNA molecule as encoded by or transcribable / transcribed from said expression cassette. The present invention further relates to method of protecting a linear deoxyribonucleic acid (DNA) molecule from exonuclease degradation by adding one or more Tus binding site(s) at the first terminus of the DNA molecule and adding one or more LacI binding site(s) at the other terminus of the DNA molecule. The present invention further relates to a method of synthesizing a protein of interest (POI) in a cell- free protein synthesis (CFPS) reaction mixture by using the (ds)DNA and / or RNA molecules of the invention, and also to a cell-free biological system or CFPS reaction mixture comprising the (ds)DNA, expression cassette and / or RNA molecules of the invention.

Aspects of the present disclosure provide methods, apparatuses, devices and systems for facilitating energy savings in a wireless network system, such as a user equipment (UE) centric –cell free (UC-CF) wireless system. By virtue of aspects of the present disclosure, network devices may be dynamically turned on / off so that energy usage may be effectively reduced at both the network side and the UE side without sacrificing performance. A network device may transmit information of a first set of cooperation network devices to the UE. The network device may determine at least one target network side device which belongs to the first set of cooperation network side devices. The network device may transmit, to the UE, an indication of the at least one target network device via lower layer signaling. The UE may establish a first connection with the at least one target network device.

Method for generating a porous injectable scaffold that includes providing a liquid composition of 2 phases at a temperature below 25° C., of Newtonian behavior, where the composition comprises:a liquid dispersant phase at room temperature formed by a gelatin with a low melting point, less than 15° C., functionalized with methacryloyl or methacrylamide groups; and a photoinitiator;and a dispersed phase, of microdroplets or beads in solid state, of a gelatin solution with a melting point greater than 25° C.;initiating the polymerization of the dispersing phase by light radiation; raising the temperature to 35-40° C. and allowing melting of the dispersed phase; and obtaining a porous scaffold.The formed porous scaffold and its use as a biological support for tissue regeneration / generation; as a biological matrix as a support for cells, for cell invasion; as an acellular biological matrix, a biological matrix as a mechanical support and / or a biological matrix for active components.

The present invention relates to a method for diagnosing cancer and predicting cancer types using a methylated cell-free nucleic acid, and more particularly, to a method for diagnosing cancer and predicting cancer types using a method for extracting methylated nucleic acids from a biospecimen, generating vectorized data of nucleic acid fragments based on aligned reads by obtaining sequence information, and then inputting the data into a trained artificial intelligence model so as to analyze a calculated value. The method for diagnosing cancer and predicting cancer types using methylated cell-free nucleic acids according to the present invention is useful because it generates vectorized data and analyzes it using an AI algorithm, compared to methods that use a conventional step of determining the amount of chromosomes based on the read count or detection methods that use the concept of distance between aligned reads to utilize values related to reads as one by one structured values, so that a similar effect can be achieved even if the read coverage is low.

The invention discloses a reagent for reducing the agglomeration rate of suspended HEK293F cells, a culture method and a culture medium. The reagent comprises the following components: 0.01-0.1% (v / v) of a nonionic surfactant, 0.05-0.2% (v / v) of polysaccharide or a derivative thereof, 1-5mM of L-glutamine or a derivative thereof, 0.001-0.01 mM of FeCl3, 14-16mM of HEPES and 50-100mu M of an RGD peptide inhibitor. According to the anti-caking agent, the nonionic surfactant, the polysaccharide or the polysaccharide derivative, the L-glutamine or the L-glutamine derivative, the FeCl3, the HEPES and the RGD peptide inhibitor are optimally proportioned, so that the HEK293F cell caking rate is remarkably reduced to 30% or below, after the anti-caking agent is added, cells are in a uniform single-cell suspension state, macroscopic cell agglomerates do not exist, and the anti-caking effect is good. The uptake efficiency of cells on nutrient substances is obviously improved. Compared with a commercially available PEG (Polyethylene Glycol)-containing anti-caking agent, the anti-caking agent disclosed by the invention has the advantages that the anti-caking rate is lower than 35% (the anti-caking rate is only 30% vs after being cultured for 48 hours, and the anti-caking rate is 40%) while the cell viability is maintained to be 95% or above, and no cytotoxic accumulation exists after long-term passage (more than or equal to 20 generations).

The present disclosure provides systems and methods to detect somatic or germline variants by providing a predetermined genomic DNA (gDNA) to an assay mixture, and capturing a sample of a subject's genetic information using a DNA sequencer and detecting genetic variants from the genetic information. A mutation may then be classified as being from a germline source if gDNA derived molecules have lengths inconsistent with those expected from cell-free DNA (cfDNA) derived molecules.

Disclosed herein are methods for processing a cell-free sample of a subject, comprising, providing said cell-free sample of said subject, wherein said cell-free sample comprises a plurality of nucleic acid molecules, subjecting said plurality of nucleic acid molecules to one or more amplification reactions to generate a plurality of complementary deoxyribonucleic acid (cDNA) molecules or derivatives thereof, and sequencing said plurality of cDNA molecules or derivatives thereof. Further disclosed herein are systems, comprising a processor, and a non-transitory computer readable storage medium encoded with a computer program that causes said processor to provide a cell-free sample of a subject, wherein said cell-free sample comprises a plurality of nucleic acid molecules, subject said plurality of nucleic acid molecules to one or more amplification reactions to generate a plurality of complementary deoxyribonucleic acid (cDNA) molecules or derivatives thereof, and sequence said plurality of cDNA molecules or derivatives thereof.

The present disclosure provides a system and method for the detection of rare mutations and copy number variations in cell free polynucleotides. Generally, the systems and methods comprise sample preparation, or the extraction and isolation of cell free polynucleotide sequences from a bodily fluid; subsequent sequencing of cell free polynucleotides by techniques known in the art; and application of bioinformatics tools to detect rare mutations and copy number variations as compared to a reference. The systems and methods also may contain a database or collection of different rare mutations or copy number variation profiles of different diseases, to be used as additional references in aiding detection of rare mutations, copy number variation profiling or general genetic profiling of a disease.

The invention provides an open type tissue culture method for plica rugosa, which comprises the following steps: S1, performing disinfection treatment on a plica rugosa explant to obtain a pretreated explant; s2, the pretreated explant is inserted into a first modified MS culture medium for first-stage culture, and a first-stage explant is obtained; s3, transferring the first-stage explant into a second modified MS culture medium for second-stage culture to obtain a second-stage explant; s4, transferring the second-stage explant into a third modified MS culture medium for third-stage culture to obtain a primary seedling. According to the method, cell-free fermentation filtrate and PPM are sequentially and progressively added into a three-stage modified MS culture medium, so that high-efficiency culture of the explant of the plica rugosa in an open environment is realized, the pollution rate and the browning degree are reduced, meanwhile, healing of the explant, adventitious bud differentiation and root primordium formation are promoted, and the seedling rate of open tissue culture of the plica rugosa is increased.

The invention provides a method for the analysis of a biological sample to determine multiple types of information therefrom in a streamlined, combined workflow, where all information is obtained in a sequencing-based analysis. The information includes the presence and concentration of specific plasma proteins in a blood sample: the number, location, and types of histone modifications associated with cell-free DNA obtained from the same sample: the sequence of cfRNA and cfDNA in the cell-free DNA sample; and epigenetic information pertaining to the cell-free DNA, such as hydroxy methylation and methylation profiles, i.e., the distribution of 5-hydroxymethylcytosine (5hmC) and 5-methylcy tosine (5mC) residues, respectively. The invention additionally pertains to a classical sequencing-based method for analyzing a biological sample to determine one or more non-classical sequence features of the sample. Compositions, kits, and related methods are also provided, including an embodiment in which truncated sequencing adapters are used in conjunction with barcoded PCR primers.

The invention relates to the technical field of bioengineering, in particular to a preparation method and application of an antibacterial peptide, and the amino acid sequence of the antibacterial peptide is shown as SEQ ID No: 1 and is named as L8. The preparation method of the antibacterial peptide L8 comprises the following steps: S1, obtaining an L8 target gene segment; connecting an L8 target gene segment with a pLC vector to obtain a connection product; transforming the connection product into an escherichia coli DH5-alpha competent cell, and screening positive clones to obtain a pLC-L8 recombinant plasmid; s2, taking the pLC-L8 recombinant plasmid as a substrate, and carrying out cell-free protein synthesis reaction to obtain the antibacterial peptide L8, the cell-free protein synthesis system comprises 175 mM of potassium glutamate, 10 mM of ammonium glutamate, 2.7 mM of potassium oxalate, 33 mM of PEP, 38 mM of compound amino acid, 10 mM of magnesium glutamate, 2% (w / v) PEG-8000, 5 nM of pLC-L8 recombinant plasmid, 11.25 mM of lactose, 33.33% (v / v) of crude extract, 100 mg / mL of escherichia coli tRNA and 2.5 nM of T7 RNA polymerase. The antibacterial peptide L8 is obtained by optimizing the antibacterial peptide LRGG, and compared with the LRGG, the antibacterial peptide L8 subjected to cell-free protein synthesis reaction expression has better antibacterial activity and biological safety.

Provided herein are cell-free compositions containing the secretome / exosomes of dsRNA primed cells ("herein dsRNA-primed secretome / exosome). The dsRNA-primed secretome is obtained from conditioned media following cell culture (for an effective amount of time) of cells treated with dsRNA, either by transfecting the cells with dsRNA or incubating the cells with dsRNA. The dsRNA-primed secretome / / exosome can be administered to a subject in need thereof, by administering an effective amount of the dsRNA-primed secretome / exosomes to a subject in need thereof. Exemplary subjects / sites of treatment include subjects include, but are not limited to in need of wound healing such as skin wound healing, burn sites, subjects in need of bone repair such as osteochondral bone repair, bone fracture sites, corneal wound sites. Other subjects include osteoporotic subjects and subjects with intervertebral disc degeneration.

Methods for the processing and analysis of blood samples obtained with blood collection tubes that reduce contamination of cfDNA by genomic DNA but that inhibit digestion by methylation-sensitive and / or methylation-dependent restriction enzymes.

The present disclosure provides methods and systems directed to cell-free identification and / or monitoring of pregnancy-related states. A method for identifying or monitoring a presence or elevated risk of a pregnancy-related state of a pregnant subject may comprise assaying a cell-free biological sample derived from said pregnant subject to detect a set of biomarkers, and analyzing the set of biomarkers with a trained algorithm to determine the presence or elevated risk of the pregnancy-related state.

Provided herein are computer-implemented methods for assigning maternal or fetal origin to one or more genetic variations in cell-free DNA (cfDNA) of a sample from a pregnant mammal, preferably a pregnant human, it uses a probabilistic model for assigning maternal or fetal origin to genetic variations in DNA from a sample obtained from a pregnant mammal, where the model assigns maternal or fetal origin based on a combination of fetal fraction and DNA fragment size.

The present disclosure provides methods and systems for screening or detecting a colorectal cancer or following colorectal disease progression that may be applied to cell-free nucleic acids such as cell-free DNA. The method may use detection of methylation signals within a single sequencing read in identified genomic regions as input features to train a machine learning model and generate a classifier useful for stratifying populations of individuals. The method may comprise extracting DNA from a cell-free sample obtained from a subject, converting the DNA for methylation sequencing, generating sequencing reads, and detecting colon proliferative cell disorder-associated signals in the sequencing information and training a machine learning model to provide a discriminator capable of distinguishing groups in a subject population such as healthy, cancer or distinguishing disease subtype or stage. The method may be used for, e.g., predicting, prognosticating, and / or monitoring response to treatment, tumor load, relapse, or colorectal cancer development.

A modified sporulating bacteria may comprise a nucleic acid cassette having a silent promotor that inhibits transcription by the modified sporulating bacteria. The nucleic acid cassette may include a nucleic acid sequence including a gene of interest that encodes a protein of interest. The nucleic acid cassette may include a nucleic acid sequence encoding a loading peptide that incorporates into a coat of a spore. The modified sporulating bacteria may comprise a pre-spore having the nucleic acid sequence having the gene of interest that encodes the protein of interest. A modified spore may be formed comprising a spore body containing the nucleic acid cassette, a spore coat, and a protein of interest derived from the gene of interest inserted into the spore coat made by exogenous cell-free biosynthesis components.

The present invention is directed to cell-free methods, such as those for determining the clostridial neurotoxin activity of a composition, determining whether or not a composition comprises clostridial neurotoxin polypeptides, and / or determining whether or not clostridial neurotoxin polypeptides or portions thereof comprised in a composition comprise an activity-altering property. The invention is also directed to an isolated capture substrate for a clostridial neurotoxin, use of the same, therapeutic or cosmetic clostridial neurotoxin compositions, and methods for producing the same.

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.

The invention provides a preparation method and application of a preparation for RNA cell-free synthesis and belongs to the technical field of molecular biology, the preparation method of the preparation for RNA cell-free synthesis mainly comprises the steps that escherichia coli serves as a chassis strain, a pET-28a carrier is utilized, a recombinant strain for expressing T7RNA polymerase is constructed, and the T7RNA polymerase has enzyme activity at the temperature of 50-60 DEG C; inducing, culturing and collecting thalli, carrying out ultrasonic lysis, and treating a lysis solution to prepare a preparation for RNA cell-free synthesis; the preparation provided by the invention naturally contains various cofactors, and RNA polymerase does not need to be extracted and purified, so that the production cost is reduced.

The present invention provides a method for screening for a candidate peptide capable of binding to a target molecule, the method including the steps of: (1) preparing a plurality of nucleic acid display libraries containing a barcoded peptide-nucleic acid complex, wherein the barcoded peptide-nucleic acid complex contains a nucleic acid moiety and a peptide moiety, the nucleic acid moiety contains a barcode sequence and a nucleic acid sequence encoding the peptide, and the plurality of nucleic acid display libraries are nucleic acid display libraries, each of which is independently produced by translation using a cell-free translation system; (2) mixing the plurality of nucleic acid display libraries to prepare a mixed nucleic acid display library; (3) bringing the mixed nucleic acid display library into contact with the target molecule; and (4) amplifying a nucleic acid corresponding to the nucleic acid moiety of the barcoded peptide-nucleic acid complex bound to the target molecule, using a barcode primer.

The invention relates to the technical field of biological tissue cell detection, in particular to an in-situ biological tissue cell free radical detection device and method based on EPR. The technical problems that in the prior art, a fluorescent probe is prone to being oxidized, the detection accuracy is affected, a traditional EPR method needs to freeze or fix a sample, a cell sample is prone to being inactivated, and free radicals in the sample are inactivated or attenuated are solved. According to the technical scheme, the in-situ biological tissue cell free radical detection device based on the EPR comprises a culture cabin, a central controller, a culture dish, a cleaning fluid storage rack, a control assembly, a detection assembly and a temperature control assembly; through the design of the movable sterile liquid suction head and the liquid conveying channel, automatic sampling and conveying are achieved, the problem that free radicals are inactivated or attenuated in traditional EPR detection is solved, the authenticity and reliability of a detection result are ensured, and real-time, dynamic and in-situ detection of biological free radicals is achieved through automatic operation of a central control system.

A method is provided for probabilistically assigning a tissue of origin to a nucleic acid in a sample, e.g., DNA in a cell-free fluid sample obtained from a human subject. A hydroxymethylation profile is generated for the sample DNA and then compared across a reference data set of hydroxymethylation profile vectors, where each hydroxymethylation profile vector identifies the hydroxymethylation profile at a specific reference locus, the tissue-specific gene associated with the reference locus, and the tissue with which the gene and reference locus are associated. A tissue of origin can be probabilistically assigned to the sample nucleic acid using the results of the comparison. Other methods of use are also provided.

The invention discloses base-modified mRNA (messenger Ribonucleic Acid) and application thereof in a cell-free protein synthesis system, and belongs to the technical field of nucleic acid modification. Natural nucleoside triphosphate is completely replaced by different types of basic group modified nucleoside triphosphate, mRNA with different chemical modifications is obtained through an in-vitro transcription method and purification, and the mRNA is put into an in-vitro cell-free protein synthesis system for reaction. A reaction element recombination system-PURE system based on prokaryote Escherichia coli purified protein and an eukaryote CFPS system taking rabbit reticulated red lysate as a material are mainly selected to detect the corresponding translation efficiency of mRNA modified by different bases in two cell-free protein synthesis systems respectively. Different regulatory activities may be shown in different cell-free protein synthesis systems when different modified nucleotides are doped into mRNA, base modified triphosphoric acid capable of improving the translation efficiency of an in-vitro protein synthesis system is screened out, and the stability of mRNA in the cell-free protein synthesis system and the corresponding protein expression level are improved.

The invention is related to materials and methods for production of tetrahydrocurcumin and tetrahydrodemethoxycurcumin from substrates. The invention provides methods and materials for cell-free and host cell-based bioproduction of tetrahydrocurcumin and tetrahydrodemethoxycurcumin.