597 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 present disclosure relates to the seminal discovery of a generation and use of genetically engineered Vibrio sp. Provided is the use of the genetically engineered bacteria for the construction, maintenance, manipulation, and / or propagation of DNA constructs; protein expression; protein secretion; vectors and other metabolic tools; metabolic engineering; expression of cellular extracts for cell-free biology; shuttle vectors; cloning vectors; and for synthetic biology applications. The disclosure also relates to the use of the replication machinery of Vibrio sp. as a cloning or expression vector for replication of recombinant DNA constructs. The disclosure also relates to methods of use of the above.

The art does not provide systematic and reproducible methods to personalize dosing of multiple immunosuppressive medications after transplantation. This invention provides a method to systematize multi-drug immuno suppression management in tissue and organ transplantation using an artificial intelligence-based complex systems approach. In embodiments of this invention, immunosuppression drug dose, blood drug concentrations, donor-derived fraction of cell free DNA (dd-cfDNA %), and aspartate aminotransferase are used to indicate allograft status or a proxy for allograft status, to generate a phenotypic response surface to produce individual treatment modalities and dosages using empirically determined unique coefficients. This surface is used to calculate appropriate immunosuppression drug doses associated with the desired outcome for that patient. Embodiments of this disclosure are directed to identifying optimized combinations of inputs for the complex system of the immunosuppressed transplant patient in order to avoid transplant rejection while avoiding unnecessary toxicity and maintaining a robust enough immune response to fight infection.

The invention discloses high-purity recombinant collagen as well as a preparation method and application thereof, the recombinant collagen is composed of a recombinant collagen monomer, and the recombinant collagen monomer has an amino acid sequence as shown in any one of SEQ ID NO: 1-22. According to the present invention, the easy-to-break site and the hydrophobicity of the collagen are optimized, such that the obtained recombinant collagen monomer and the prepared recombinant collagen have characteristics of high cell activity (adhesion, migration and proliferation), high triple helix structure stability, high purity, no cytotoxicity, large-scale production, and production cost reduction.

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.

Provided herein are methods for replenishing dissolved gas to aqueous droplets on a digital microfluidic device. Provided herein are methods of cell-free protein synthesis, optimized cell-free protein synthesis (CFPS) reagents, and methods for optimizing CFPS reagents to increase protein expression yield. The method is suitable for protein expression on microfluidic devices having hydrophobic surfaces by circulating a fill fluid to introduce additional oxygen.

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.

Disclosed herein in some embodiments are methods, compositions, and systems for distinguishing between ulcerative colitis (UC), Crohn's disease (CD) and other Inflammatory Bowel Disorders (IBD) by sequencing cell free nucleic acids. In some embodiments, microbial cell-free nucleic acid sequencing can provide data that can determine whether UC, CD, or other IBD are asymptomatic, in remission, or active. In some embodiments, microbial cell-free nucleic acid sequencing can provide data that can determine whether an active form of UC, CD, or other IBD is mild, moderate, or severe.

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.

A method of identifying a variant as a somatic variant derived from cell free DNA (cfDNA) identifies the variant at a locus based on differences between the nucleic acid sequence for a cfDNA fragment in a plurality of cfDNA fragments and a nucleic acid sequence for the locus in a reference sequence, where the cfDNA fragments are from a liquid biopsy sample from a subject. A set of cfDNA fragments comprising the variant, in the plurality of cfDNA fragments, determines fragment length metrics. A variant allele fraction (VAF) is determined based on comparison of the number of cfDNA fragments having the variant and the total number of cfDNA fragments mapping to the locus. Clonal hematopoiesis prevalence metrics for the variant are obtained. The fragment length metrics, VAF, and hematopoiesis metrics are inputted into a model thereby obtaining, as model output, whether the variant is a somatic variant derived from cfDNA.

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.

The present disclosure provides a method for the preparation and analysis of a biological sample of a xenograft receptor wherein the method comprises extracting fragmented or intact cell-free DNA, RNA (such as mRNA or miRNA) or protein of a sample derived from the xenograft receptor, and measuring the amount of cell-free DNA, RNA (such as mRNA or miRNA) or protein derived from said xenograft enables the assessment of xenograft rejection. The detection of the cell-free DNA or RNA can be carried out by preparing a sequencing library and carrying out whole genome sequencing.

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.

Provided is a method and apparatus for cell staining without cell loss, and more particularly, to a method and apparatus for cell staining without cell loss during treatment of a staining reagent or washing reagent by immobilizing cells to be analyzed in a phase change material before staining. A method for cell staining without cell loss may effectively prevent cell loss during staining and analysis of rare cells that are becoming important in clinical diagnosis, and thus may facilitate observation, analysis and diagnosis thereby even with a trace amount of sample. In addition, as the method for cell staining without cell loss allows immobilization of cells, it makes discrimination and isolation of single cells easy, and may be effectively used in analysis of the isolated cells.

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 present invention discloses a method for cancer pre-screening of a subject. Genetic risks associated with genetic characteristics are determined by sequencing and analyzing cell free DNA ("cfDNA") fragments present in a blood sample of the subject. The clinical risk is determined based on factors such as age, gender, and race. In some cases, clinical factors specific to certain cancers, such as smoking conditions, are combined. By incorporating clinical risks into genomic risk analysis, improved lung cancer pre-screening results are provided, enabling the same number of positive cancer detections to be achieved using a small number of LDCT lung cancer screening times.

The present invention provides a large Stokes shift near-infrared fluorescent probe for detecting HClO, its preparation method and biological imaging application, belonging to the field of bioanalysis technology. Based on the strategy of modifying the substituents at the 3,6 positions of xanthene, the present invention constructs a large Stokes shift near-infrared fluorescent probe for specifically recognizing HClO. Optical experiments show that the emission wavelength of the probe for detecting HClO is 776 nm, the Stokes shift reaches 148 nm, with strong selectivity, high sensitivity, and a detection limit as low as 4.95 nM, which can greatly reduce the interference of background fluorescence in biological imaging applications. The fluorescent probe of the present invention has almost no cytotoxicity and can be applied to the detection and imaging of HClO in living cells MCF-7 and rat liver tissues, with a tissue penetration depth of 140 μm.

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.