23 results about "A deoxyribonuclease" patented technology

The present application relates to a deoxyribonuclease mutant and its application in preparing PDRN. The amino acid sequence of the deoxyribonuclease mutant is shown as SEQ ID NO. 1, and the nucleotide sequence of the encoding gene is shown as SEQ ID NO. 2. The present application also provides the application of the deoxyribonuclease mutant and / or the encoding gene of the deoxyribonuclease mutant in preparing PDRN, and the specific application method. The present application is based on the deoxyribonuclease expressed by the DNASE1 gene of bovine pancreas tissue, and the amino acids at positions 69, 132 and 215 are selected for iterative saturation mutation. Compared with the wild-type enzyme, the enzyme activity of the deoxyribonuclease mutant A69H-S132R-Q215C is increased by 2.15 times, reaching 4.1x10 6 U / mL, and the DNA extraction rate of salmon PDRN prepared by using the mutant enzyme prepared by the present application is 15.2%, the DNA content is 99.1%, and the molecular weight is 1.05 million Daltons, which is significantly improved compared with the wild-type enzyme, and the PDRN in this molecular weight range has more significant biological activity.

The application provides a kit for extracting RNA and application thereof. In the application, deoxyribonuclease (DNase) is used for DNA digestion in the process of magnetic bead extraction. The reaction environment can meet the optimal reaction environment requirement of the deoxyribonuclease (DNase), and at the same time, a hydrophobic environment is provided when the magnetic beads are adsorbed, so as to ensure the yield of nucleic acid. In addition, an emulsion of an oil is added to increase the total volume during the extraction operation, so as to facilitate the convenience of automatic extraction operation of the instrument. The extracted RNA has high yield and high purity, and the method can easily realize automatic extraction of the instrument. The requirement of extracting high-purity RNA quickly and conveniently in the process of clinical practical application is solved, the difficulty and cost of extracting high-purity RNA are greatly reduced, and the extracted RNA can be well adapted to subsequent RT-PCR and RNA-seq application.

Abstract Described herein, in one aspect, are antigen presenting cells (APCs) comprising an exogenous nucleic acid encoding one or more candidate antigens, wherein the one or more candidate antigens are expressed and presented with MHC class I or MC class II molecules; a molecular reporter of Granzyme B (GzB) activity; and c) an exogenous inhibitor of caspase-activated deoxyribonuclease (CAD)-mediated DNA degradation, a CAD knockout, or a caspase knockout (e.g., caspase 3 knockout). Described herein, in another aspect, is a system for detection of recognized antigen presentation by an antigen presenting cell to a cytotoxic lymphocyte or NK cell. Abstract 2018 / 22761 oM - cell Target ml Target cell cell cell Target Target Target SUBSTITUTE SHEET (RULE 26) cell cell cell Target Target cell cell 1 / 28 my Isolate recognized cell Library of target cells target cells and displaying different Add T cells from sample sequence antigens antigens of interest. CTLs deliver cytotoxic granules to target cells displaying cognate antigen FIG. 1 PCT / US2018 / 036663 20 26 20 53 56 07 J ul 2 02 6 2 0 2 6 2 0 5 3 5 6 0 7 J u l 2 0 2 6 2 0 1 8 / 2 2 7 6 1 o M a n d m y 1 / 2 8 m y L i b r a r y o f t a r g e t c e l l s d i s p l a y i n g d i f f e r e n tA d d T c e l l s f r o m s a m p l e of interest. CTLs deliver c y t o t o x i c g r a n u l e s t o t a r g e t c e l l s d i s p l a y i n g c o g n a t e a n t i g e n P C T / U S 2 0 1 8 / 0 3 6 6 6 3

The present invention provides a method for reducing the amount of DNA remaining in a biopolymer so as to increase the purity of the biopolymer when producing the biopolymer by means of a microorganism. This biopolymer production method includes: an endodeoxyribonuclease treatment step for bringing an endodeoxyribonuclease into contact with biopolymer granules obtained by culturing a microorganism capable of producing a biopolymer; and a pH adjustment step for keeping the biopolymer granules under the condition of a pH of 7.0 or higher at least at one time point after the start of said endodeoxyribonuclease treatment.

The present invention provides compositions that modulate gut microbiota and / or the activity of deoxyribonucleases (e.g., in the gastrointestinal tract) and are useful in the treatment of diseases and disorders related to the activity of deoxyribonucleases including, for example, neurodegenerative, inflammatory, and metabolic diseases and cancer.

The application discloses a kind of nanoparticles for removing lung mucus and its preparation method and application, it is related to the field of biological medicine.The nanoparticles of the application are composed of inner core and shell coated on the surface of the inner core, wherein the inner core contains citric acid and photoluminescent polymer-aliphatic polyester copolymer;The shell contains deoxyribonuclease and hydrophilic polymer.The nanoparticles provided by the application have excellent biocompatibility and multifunctional integration characteristics, which realizes effective penetration of lung mucus barrier, specific degradation of DNA network, down-regulation of pro-inflammatory factors and inhibition of mucus production in sequence through cascade response mechanism, breaking through the limitations of traditional single therapy.In addition, the in vivo experimental results show that it can significantly reduce the thickness of lung mucus and the protein concentration of lavage fluid, and effectively relieve airway obstruction, and can be used for removing lung mucus.

The invention provides a preparation method of decellularized omentum majus, and relates to the technical field of biomedical materials.The method comprises the steps that in a closed container, omentum majus subjected to freeze-drying treatment is placed in a mixed solvent to be subjected to degreasing treatment, and a first intermediate is obtained; performing vacuum pulse treatment on the closed container in the degreasing treatment process; cleaning the first intermediate to obtain a second intermediate; performing circulating oscillation treatment on the second intermediate for at least one time to obtain a third intermediate; the third intermediate is cleaned in a phosphate buffer solution and then placed in a compound enzyme solution containing pancreatin and deoxyribonuclease I to be subjected to oscillation treatment, and a fourth intermediate is obtained; the fourth intermediate is sequentially washed with deionized water and a phosphate buffer solution and then placed in a sucrose solution to be soaked, then freeze-drying and sterilization are sequentially conducted, and the decellularized omentum majus is obtained. By adopting the method disclosed by the invention, the decellularized omentum majus with high purity, low residue and complete structure can be obtained.

Some embodiments of the present disclosure relate to one or more compositions that upregulate the production of one or more sequences of mRNA. The sequences of mRNA may encode for translation of a target biomolecule, thereby causing an increase in bioavailability of the target biomolecule within a subject that is administered the one or more compositions. In some embodiments of the present disclosure, the target biomolecule is a fusion protein with an Fc fragment, such as a toll-like receptor 3-Fc (TLR3-Fc). In some embodiments of the present disclosure, the target biomolecule is toll-like receptor 9-Fc (TLR9-Fc). In some embodiments of the present disclosure, the target biomolecule is deoxyribonuclease I-Fc (DNAse I-Fc). In some embodiments of the present disclosure, the target biomolecule is neural growth factor-Fc (NGF-Fc). In some embodiments of the present disclosure, the target biomolecule is insulin-Fc.

The present invention provides compositions that modulate gut microbiota and / or the activity of deoxyribonucleases (e.g., in the gastrointestinal tract) and are useful in the treatment of diseases and disorders related to the activity of deoxyribonucleases including, for example, neurodegenerative, inflammatory, and metabolic diseases and cancer.

This invention belongs to the medical field and provides a calcium carbonate manganese nanoprobe for in situ imaging detection of vascular epidermal growth factor, its preparation method, and its application. In this strategy, a DNA strand (P1 and P2) containing a protein recognition antibody, a protein recognition sequence, and a deoxyribonuclease, along with a molecular beacon DNA strand (H1), is stirred in a DMEM cell culture medium containing calcium chloride and manganese chloride to prepare the calcium carbonate manganese nanoprobe. In acidic tumor cells, this nanoprobe releases P1, P2, H1, and manganese ions. P1 and P2 can simultaneously recognize proteins, forming a complete deoxyribonuclease center, which catalyzes the cleavage of H1 under the action of the cofactor manganese ions, generating a fluorescent signal. This strategy achieves specific in situ dynamic imaging of low-abundance proteins in vivo and has great potential in biomedical research.

Therapeutic compositions for treating a subject, the compositions including a polymeric nanoparticle, an active agent encapsulated within the polymeric nanoparticle, and a delivery vehicle for targeted delivery of the polymeric nanoparticle and encapsulated agent to a target tissue in the subject. The therapeutic compositions can be designed for targeting the destruction of a Nuclei neutrophil extracellular trap (NET) in a venous thrombus in a subject. The active agent can be an enzyme, including deoxyribonuclease 1 (DNase 1) enzyme. Methods of treating a subject using the therapeutic compositions are provided, including methods of treating venous thrombus, deep vein thrombosis, or a related blood clotting disease and / or condition.

Some embodiments of the present disclosure relate to one or more compositions that upregulate the production of one or more sequences of mRNA. The sequences of mRNA may encode for translation of a target biomolecule, thereby causing an increase in bioavailability of the target biomolecule within a subject that is administered the one or more compositions. In some embodiments of the present disclosure, the target biomolecule is a fusion protein with an Fc fragment, such as a toll-like receptor 3-Fc (TLR3-Fc). In some embodiments of the present disclosure, the target biomolecule is toll-like receptor 9-Fc (TLR9-Fc). In some embodiments of the present disclosure, the target biomolecule is deoxyribonuclease I-Fc (DNAse I-Fc). In some embodiments of the present disclosure, the target biomolecule is neural growth factor-Fc (NGF-Fc). In some embodiments of the present disclosure, the target biomolecule is insulin-Fc.

The application provides a PDRN and an extraction method and application thereof, and belongs to the technical field of PDRN extraction. The extraction method of the PDRN comprises the following steps: subjecting plant tissues to stress treatment, so that the cells are in an irreversible damage state; subjecting the plant tissues subjected to the stress treatment to incubation treatment, so that DNA is selectively partially degraded under the action of endogenous deoxyribonuclease; and subjecting the plant tissues subjected to the incubation treatment to crushing treatment and extraction and purification treatment, to obtain the PDRN. The extraction method selectively degrades DNA under mild conditions by using endogenous enzymes (such as DNase) of plants, and has the advantages of high yield, high efficiency, low cost and green environmental protection; meanwhile, the PDRN extracted by the extraction method also has the advantages of high purity, high physiological activity and high safety.

The invention discloses a sensor for detecting cadmium, the sensor comprises a reference electrode, a counter electrode, a working electrode and an electrolyte, the working electrode comprises a substrate, the surface of the substrate is coated with nanogold modified graded porous carbon Au-coated HPC, the surface of the substrate is modified with an oligonucleotide chain sS1, and after reaction, the obtained working electrode is HCR / rS1 / MCH / H1 / Au-coated HPC / substrate. According to the sensor for detecting cadmium, the sensor is prepared by combining Cd < 2 + >-dependent deoxyribonuclease (DNAzyme), hybrid chain amplification reaction (HCR) and hierarchical porous carbon (HPC). Obvious advantages are shown in key performances such as sensitivity and detection range. The method is used for detecting national standard substances of rice and soil, the result is well within a marked range, the adding standard recovery rate ranges from 98.4% to 107.0%, the method has good consistency with the detection result of an atomic absorption graphite furnace (AAS) method, and the method has good application value in ultrasensitive accurate analysis of Cd < 2 + > in the fields of food, environment, sanitation and the like.

The application discloses a kind of ternary co-modified engineering velvet MSCs exosomes and preparation method and application thereof, belong to the biomedicine technical field, it includes velvet MSCs exosomes, polyarginine polypeptide-deoxyribonuclease I complex located on the surface of velvet MSCs exosome, and RGD polypeptide with Arg-Gly-Asp sequence modified on the surface of velvet MSCs exosome.The purpose is to solve the problem that the "integration" treatment mode of multiple pathological links cannot be realized when existing RA is treated, for the first time, three active components polyarginine polypeptide (P-Arg), deoxyribonuclease I (Dnase I) and RGD polypeptide are modified on the surface of velvet MSCs exosome, to form surface engineered modified exosome complex, multiple functions of immune regulation, cfDNA removal, targeted delivery and tissue protection can be realized in RA lesion, and a new, systematic treatment method is provided for rheumatoid arthritis.

This application belongs to the field of bioanalytical detection technology, specifically disclosing a detection system and method for live Salmonella based on deoxyribonuclease activation. The detection system includes a Sub probe, a deoxyribonuclease probe, a hairpin probe, hyperbranched dendritic nanomolecules, a crRNA / Cas12a binary complex, a fluorescent probe, and magnetic beads. The hyperbranched dendritic nanomolecules include substrate A, substrate B, and a trigger strand B probe. Substrate A consists of an A-F strand probe, an A-Q strand probe, and a helper strand A probe, while substrate B consists of a B-F strand probe, a B-Q strand probe, and a helper strand B probe. This detection system specifically recognizes ribonuclease H2 released only by the metabolism of live Salmonella and utilizes it to activate the Sub-Dz substrate, ensuring from the source that only live bacteria can trigger the subsequent reaction. This effectively solves the problem of interference from dead bacteria, achieving highly sensitive, highly specific, and rapid detection of live Salmonella.

The present invention relates to a cfDNA standard and a preparation method therefor. The method comprises the following steps: a1) constructing a vector expressing DNA fragmentation factor subunit beta (DFFB) or deoxyribonuclease 1 like 3 (DNASE1L3); a2) culturing cells to be transfected; b) transfecting a vector having a DFFB gene and a vector having a DNASE1L3 gene into said cells and culturing same; and c) for the successfully transfected cells, collecting culture supernatant and extracting cfDNA.

A method for collecting cells from a bioreactor is provided wherein the bioreactor comprises cells to be collected, at least one inlet and / or outlet, and a culture chamber in which the cells are attached to a matrix. The method comprises providing a wash solution having a cell dissociation reagent (CDR) into the culture chamber; and after providing the washing solution, removing and capturing the detached cells from the culture chamber via the at least one inlet and / or outlet. The cell dissociation agent includes an endonuclease, in particular a deoxyribonuclease, such as deoxyribonuclease I (DNase I). The wash solution may further include a divalent metal ion, such as Mg + 2, Ca + 2, or Mn + 2.

The application belongs to the technical field of biological medicine, and particularly relates to a thymus tissue dissociation reagent, a kit and application thereof in obtaining thymus epithelial cells. The thymus tissue dissociation reagent at least comprises collagenase D, dispersinase II, deoxyribonuclease I, dissociation enzyme and a dissolving reagent; and collagenase II and / or collagenase IV and / or trypsin. The cell survival rate of the thymus tissue dissociation reagent after dissociating the thymus tissue is maintained at a high level of 95.7% to 99.6%, which is significantly higher than that of the prior art; and after sorting, the number of thymus epithelial cells enriched and obtained from each gram of thymus tissue is 1.38e 5 ~1.06e 6 , and the success rate of enriching and obtaining thymus epithelial cells is as high as 95%, which is significantly higher than that of the prior art; and the proportion of epithelial stem cells in the thymus epithelial cells is 65.7%, which is significantly higher than that of the prior art.

The inventors have now succeeded in efficiently conjugating clusters of ultrasmall particles of iron oxide (USPIO) and polycathecolamine or polyserotonine with an activator of fibrinolysis, in particular a tissue plasminogen activator (tPA). Such conjugates are useful for detecting microthrombi in a condition requiring thrombolysis, in particular acute ischemic stroke, by magnetic resonance imaging (MRI), and for treating a condition requiring thrombolysis, in particular acute ischemic stroke. The present invention relates to conjugate comprising a suspension of biocompatible particles comprising ultrasmall particles of iron oxide embedded in a polycathecolamine or polyserotonine matrix, an activator of fibrinolysis or a fragment thereof and optionally a deoxyribonuclease, and its use in an in vivo method of detection of microthrombi in a condition requiring thrombolysis, in particular acute ischemic stroke, by magnetic resonance imaging (MRI) and in the treatment of a condition requiring thrombolysis, in particular ischemic stroke.

Some embodiments of the present disclosure relate to one or more compositions that upregulate the production of one or more sequences of mRNA. The sequences of mRNA may encode for translation of a target biomolecule, thereby causing an increase in bioavailability of the target biomolecule within a subject that is administered the one or more compositions. In some embodiments of the present disclosure, the target biomolecule is a fusion protein with an Fc fragment, such as a toll-like receptor 3-Fc (TLR3-Fc). In some embodiments of the present disclosure, the target biomolecule is toll-like receptor 9-Fc (TLR9-Fc). In some embodiments of the present disclosure, the target biomolecule is deoxyribonuclease I-Fc (DNAse I-Fc). In some embodiments of the present disclosure, the target biomolecule is neural growth factor-Fc (NGF-Fc). In some embodiments of the present disclosure, the target biomolecule is insulin-Fc.

This invention discloses a mineralization enzyme composite powder for disrupting biofilms, its preparation method, and its antibacterial applications. The composite powder is a hybrid nanoparticle prepared by a one-step biomineralization process. The composite powder contains nucleases and bactericidal enzymes; the nucleases are one or more of deoxyribonucleases, ribonucleases, or non-specific nucleases in any proportion; the bactericidal enzymes are single or multiple enzymes, wherein the multi-enzyme cascade generates hydroxyl radicals. This invention achieves a synergistic effect of biofilm degradation and antibacterial activity by integrating multiple natural enzymes. It not only significantly improves the stability of nucleases but also enhances the cascade catalytic efficiency of the bactericidal enzymes, thereby effectively disrupting biofilms and increasing the generation efficiency of hydroxyl radicals. This invention also provides the antibacterial applications of the composite powder in vitro and in vivo, exhibiting good stability and catalytic performance, suitable for various anti-infection scenarios.

The invention relates to methods for immunomodulation of immunosuppressive tumor cell microenvironment and prevention of tumor microbiome effects through multiple pathways with DNase enzyme therapy. Methods for immunomodulation of immunosuppressive tumor cell microenvironment comprising administering an effective amount of deoxyribonuclease (DNase) enzyme, either alone or in combination with other immunomodulating agent.