277 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 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.

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 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.

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.

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.

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 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.

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.

The application provides a kind of mitochondrial function optimization additive for cell culture, preparation method and application, belongs to cell culture technical field.The additive includes core active ingredient, auxiliary component and carrier, and the core active ingredient includes D-Arg-Cha-Lys-Phe-NH2 Polypeptide, ubiquinone-10 phosphate, reduced glutathione and alpha-lipoic acid.The application forms a multi-target core active system through the synergy of the four, realizes the triple synergistic protection of stabilizing mitochondrial cardiolipin, maintaining mitochondrial morphology and membrane potential, and reducing mitochondrial ROS, which can significantly improve cell culture quality.When applied, the mitochondrial cardiolipin content can be increased by more than 50%, the mitochondrial abnormality ratio can be reduced to less than 12%, the membrane potential positive cell ratio can be increased to more than 86%, the mitochondrial ROS level can be reduced by more than 40%, and the cell proliferation rate can be increased by more than 25%, which is suitable for in vitro culture of various eukaryotic cells, and the preparation process is simple and has no cytotoxicity.

Methods are provided for detecting gene deletions and driver mutations using tumor-specific methylation patterns from cell-free DNA and tissue biopsies. The methods exploit the mutual exclusivity between tumor-specific methylation and homozygous gene deletion, enabling detection through absence of expected methylation signals. Applications include detection of MTAP, PTEN, and RB1 deletions, as well as prediction of EGFR single nucleotide (SNV) and small (up to 50 base pair) insertions and deletions (indel) driver mutations, enabling identification of patients eligible for targeted therapies.

Disclosed herein are methods and systems for targeted detection of circulating tumor DNA (ctDNA) molecules. In some cases, a methylated DNA depleted molecular sequencing library can be generated and used to reliably detect ctDNA in cell-free DNA samples at lower sequencing depths and at lower costs than existing methods.

This invention discloses a method for the synergistic metabolic analysis of multiple components of traditional Chinese medicine (TCM) integrated with the gut microbiota. After initializing the microbiota ecosystem, a solution of multi-component TCM extracts is continuously injected into a microfluidic chip. Stable isotope-labeled precursors are pulsed into a specific simulation region to perform spatial isotope encoding. A standing wave acoustic field is applied at the sampling node of the microfluidic chip, and multiphase fluid spatial splitting is performed using the difference in the primary acoustic radiation force experienced by the particles to derive cell-free TCM metabolites. The cell-free TCM metabolites are separated by chromatography and gas phase drift separation. A four-dimensional metabolic feature array is constructed, and the absolute value of the mass shift is calculated for spatial inverse decoding. Closed-loop feedback is triggered based on the rate of change of target metabolite ion abundance, increasing the amplitude of the standing wave acoustic field pressure to confine intestinal bacteria and performing in-situ enrichment of synergistic metabolic intermediates. This invention realizes the spatial metabolic trajectory tracing and network reconstruction of multiple components of TCM in a complex gut microbiota ecosystem.

The present invention provides a method and means for isolating or recovering short-chain DNA such as cell-free DNA (cfDNA) from a blood sample. Specifically, the present invention provides a method for isolating or recovering short-chain DNA of 100-250 bp from a DNA sample, the method comprising: a step for introducing the DNA sample into a column equipped with an ultrafiltration filter and centrifuging the DNA sample; and a step for recovering a filtrate containing the short-chain DNA.

Aspects of the present disclosure includes methods to quantify methylation in a DNA sample, where the method includes treating the sample to encode the presence or absence of DNA methylation, adding to the sample a set of synthetic molecules (e.g., quality control template (QCT) molecules), generating a co-amplification mixture, sequencing the co-amplification mixture, determining a number of methylated molecules in the sample based on the number of methylated reads from the sample and a number of reads from the set of synthetic molecules.

Provided are engineered polypeptides having dehydrogenase activity that exhibit increased catalytic efficiency for nicotinamide-based unnatural cofactors relative to wild-type or parent polypeptides that encode a dehydrogenase that utilizes nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate. Also provided are cell-free and whole cell biotransformation systems for converting a substrate into a product using a redox reaction with a re-cycled unnatural cofactor that utilizes an engineered polypeptide that has dehydrogenase activity and which exhibits increased catalytic efficiency for nicotinamide-based unnatural cofactor.

A method of sequencing a nucleic acid of interest (NAOI) is provided. In some embodiments, the method may comprise providing a sample from a patient, the sample having cell-free DNA molecules comprising NAOIs, amplifying the NAOIs, ligating adapters to the NAOIs, and sequencing the NAOIs. A method of labelling a NAOIs is also provided. In some embodiments, the method may comprise providing a sample from a patient having the NAOIs, amplifying the NAOIs, contacting the amplified NAOIs with a pool of oligonucleotides, attaching oligonucleotides to each of the amplified NAOIs to label the NAOIs. The method may further comprise sequencing the labelled NAOIs and grouping the resulting sequencing reads.

Compositions and methods disclosed herein relate to improved in vitro cell-free systems for various biological reactions. In one aspect, a composition of the present disclosure includes: a cell-free extract derived from an organism; a nucleic acid; and an organelle that is heterologous to the organism.

Provided herein are methods of cell-free protein synthesis, optimized cell-free protein synthesis (CFPS) reagents, and methods of optimizing CFPS reagents to increase protein expression yield using additional NTPs and / or additional energy sources added with nucleic acid templates. The methods are suitable for protein expression on microfluidic devices having hydrophobic surfaces by combining droplets on the device for parallel screening of a series of expression compositions.

The invention relates to the field of biological medicine, and discloses application of an acellular fat extract in preparation of a medicine for accelerating chronic wound healing. Researches show that CEFFE can improve healing of chronic wounds of diabetic mice. This is achieved by the pro-angiogenesis and anti-inflammatory activity of CEFFE. The existence of various growth factors in the CE FFE can explain the influence of the growth factors on the functions of epidermal cells, endothelial cells, fibroblasts and macrophages. CEFFEE may be used as a therapeutic drug for treating chronic wounds of diabetic patients, and is expected to improve the overall prognosis of the diabetic patients and improve the life quality of the patients.

The present disclosure provides methods for detecting the presence or absence of cancer in a subject. In some embodiments, samples are obtained from the subject, and a panel capable of detecting multiple biological molecules, such as RNA, cDNA, and cell-free DNA (cfDNA) is utilized to processing a ample to determine tumor variants in the first sample. A mixture of these biological molecules are detected via oligonucleotide probes in order to detect as a particular sensitivity within a sequencing read budget, further including exon related probe designs for RNA.

The present disclosure includes, among other things, methods, kits, and systems for determining a status of lung cancer. In various embodiments, the present disclosure relates to the use of one or more histone modifications, chromatin accessibility, binding of one or more transcription factors, and / or DNA methylation as features of a status of lung cancer. In some embodiments, differential modifications and / or differential accessibility are detected and quantified at one or more genomic loci in a biological sample, e.g., cell-free DNA (cfDNA), from a liquid biopsy sample obtained or derived from a subject having lung cancer. In various embodiments, the determined status can be used, e.g., to select a treatment for lung cancer and / or to treat lung cancer.