28 results about "Enzymic digestion" patented technology

A system for targeted delivery of a medicament to the gastrointestinal tract (e.g., selectively to the duodenum) of a subject. The system includes a medicament, a mucoadhesive layer at least partially surrounding the medicament, and an enteric coating encasing the medicament and mucoadhesive layer. The enteric coating comprises a material degradable by a pH of the GI tract and a material digestible by a lipase. Also provided are method of making and using the system for targeted delivery of a medicament to the gastrointestinal tract. Also provided are a system for deploying a medicament-containing needle system using an expanding element. The system includes a medicament-containing needle system and an expanding element within a capsule. The capsule is initially coated with an enteric coating. As the enteric coating degrades the expanding element absorbs fluid that enters the capsule and expands to deploy the medicament-containing needle system from the capsule.

The disclosure relates to methods of detecting Host Cell Proteins (HCPs) in biopharmaceutical samples containing a drug product. More specifically, the disclosure relates to methods of assaying HCPs in biopharmaceutical samples using enzymatic digestion protocols, liquid chromatography (LC) gradients, immunoaffinity capture protocols, and High-Resolution Mass Spectrometry (HRMS) to enhance the recovery, detection, and profiling of HCPs, which can impact the efficacy, shelf-life, and therapeutic response of drug products.

The invention discloses a culture medium for identifying gardnerella vaginalis, and relates to the technical field of bacterial culture media, the key points of the technical scheme are as follows: the culture medium comprises the following raw material components by concentration: 10.0 g / L of casein pancreatin digest, 3.0 g / L of cardiac pancreatin digest, 5.0 g / L of yeast extract powder, 1.0 g / L of corn starch, 5.0 g / L of sodium chloride and 13.0 g / L of agar, and the final concentration of defiberized sheep blood is 5%. The pharmaceutical composition is prepared from 1 mg / mL of sodium alginate, 1 mg / mL of sodium alginate, 1 mg / mL of polyanethole Sulfonate (SPS) and 10 [mu] g / mL of vancomycin. According to the invention, through optimal combination of specific components and dosages thereof, rapid identification and screening of gardnerella vaginalis can be realized, and the method has the advantages of accurate result, strong specificity and the like, and has practical popularization and application values.

This invention relates to the interdisciplinary field of cell biology, experimental oncology, and translational medicine, specifically to a cancer-associated fibroblast cell line, its construction method, and its applications. This invention fills the research gap in mouse-derived oral squamous cell carcinoma-associated fibroblast cell line models and provides a method for constructing such a cell line. Through the inverted adherent culture and enzymatic digestion methods of this invention, a mouse-derived oral squamous cell carcinoma-associated fibroblast cell line with a clear origin, high purity, and long-term phenotypic and functional stability can be obtained. This allows for the establishment of a cryopreservable and resuscitable mouse oral squamous cell carcinoma-associated fibroblast cell bank, providing technical support for research related to the tumor microenvironment and immunotherapy.

Described herein are methods for enriching, identifying, quantifying, and / or characterizing a protein of interest in a complex biological sample, such as a serum sample. The protein of interest may be enriched using a library of bead-based peptide ligands. The enriched protein of interest may subsequently be subjected to denaturation, reduction, and / or enzymatic digestion, to generate peptides which can be detected using liquid chromatography-mass spectrometry (LC-MS) techniques, and which are uniquely identifying of the protein of interest.

This invention discloses a method for producing single-stranded DNA using Cas nickase. This method solves the problem of limitations on the enzyme cleavage sites when using nicking endonucleases, and improves the nicking efficiency of plasmids by performing multiple nicking using multiple gRNAs in a single enzyme cleavage system, while also effectively improving the efficiency of subsequent digestion by exonucleases.

The invention relates to a propionibacterium acnes selective culture medium and a preparation method thereof, the culture medium comprises casein pancreatin digest, meat and stomach enzyme digest, heart and pancreatin digest, yeast extract powder, corn starch, sodium chloride, agar and defiberized sheep blood, and the volume is fixed with distilled water. Compared with an existing culture medium, the culture medium has the advantages of being reliable in detection and high in propionibacterium acnes separation rate and has a good application prospect.

The invention provides inter alia a method for enzymatic digestion of an extracellular matrix protein in a tissue sample, comprising the steps of:—providing an aqueous medium that comprises (i) a matrix metalloproteinase and (ii) a cation that is Ca2+ at a concentration of at least 2 mmol / L;—contacting said aqueous medium with a tissue sample that comprises an extracellular matrix protein under conditions that allow for enzymatic digestion of said extracellular matrix protein; wherein said tissue sample is a cartilage tissue sample.

The invention relates to the technical field of drug detection, in particular to immobilized protease K and application thereof in hair drug detection. The immobilized protease K comprises zirconia ceramic balls and protease K which is covalently connected with the zirconia ceramic balls through APTES amination; wherein the loading capacity of the protease K is 1-5mg / g. The specific detection method comprises the following steps: carrying out milling and enzyme digestion on to-be-detected hair and the immobilized protease K zirconia ceramic ball, and carrying out fluorescence immunochromatography on the obtained supernatant to detect the narcotics and metabolites thereof. According to the application, covalently solidified protease K is adopted, the operation stability is excellent, 50 DEG C high temperature and mechanical oscillation can be resisted, 85-95% efficient release of traditional difficult samples such as weak positive hair and dark hair can be realized, the detection rate and the detection accuracy of drugs and metabolites thereof are improved, and the false negative risk is reduced; the requirements of laboratory high-throughput detection and on-site rapid screening are met.

The present disclosure relates to a method of determining the presence of at least one messenger RNA (mRNA) molecule in a composition, the method comprising: (a) reverse transcription of the at least one mRNA molecule to obtain a template comprising at least one first strand of complementary DNA (cDNA); (b) generating at least one double-stranded cDNA molecule from the template; (c) amplifying the at least one double-stranded cDNA molecule; (d) digesting the at least one amplified double-stranded cDNA molecule with at least one restriction enzyme to obtain a test cDNA fragment; (e) separating the test cDNA fragments so as to form a test cDNA fragment map; and (f) comparing the test cDNA fragment map to a control cDNA fragment map wherein the at least one mRNA molecule is present in the composition when the test cDNA fragment map comprises the control cDNA fragment map. Also disclosed are methods of quantifying and / or assessing the integrity of at least one mRNA molecule in a composition, processes of making a composition comprising at least one mRNA molecule and a primer pair for use in the methods of the invention.

The present disclosure provides a method and composition for preparing a nucleic acid library.In some embodiments, the nucleic acid comprises cell-free DNA, including cfDNA that requires analysis, for example, by sequencing.The method can include restriction enzyme digestion, adapter ligation, and subsequent amplification, and can provide an improved approach to reduce the number of adapter-dimers generated during the process.In some aspects, the method for preparing a nucleic acid library can include: digesting DNA molecules with restriction enzymes to generate DNA fragments; ligating adapters to the DNA fragments by incubating with ligase to generate a mixture of adapter-ligated DNA fragments and adapter-dimers; amplifying the adapter-ligated DNA fragments to generate amplified adapter-ligated DNA fragments; and reducing the amount of adapter-dimers by distinguishing between the junction between adapter and DNA fragment and the junction between adapter and another adapter. TIFF2022530289000002.tif98128

To enable high-sensitivity detection of peach that might cause allergies contained in food ingredients and products even if the amount is minute.SOLUTION: A method of detecting peach is provided, comprising extracting protein from a sample, obtaining an enzyme digest of the extracted protein using a protease, and analyzing the enzyme digest to detect at least one peptide selected from a group consisting of SEQ ID NOs: 1 through 3 using a mass spectrometer so as to qualitatively or quantitatively determine the presence or absence of peach protein in the sample.SELECTED DRAWING: Figure 1

Preparation of a product comprising stromal vascular fraction cells includes washing human biological material comprising adipose in a container apparatus having an internal filter, which 5 divides an internal containment volume of the container apparatus into a tissue retention volume on one side of the filter and a filtrate volume on an opposite side of the filter, and a mixing device with at least one rotatable mixing member disposed in the tissue retention volume. The washing includes operation of the mixing device to rotate the mixing member through the human biological material within the tissue retention volume, and the washing is followed by digesting washed material within 10 the internal containment volume with added enzyme, centrifuging of the container apparatus to prepare a centrifuged pellet in the filtrate volume, selectively removing material of the pellet and preparing a product with a mixture of stromal vascular fraction cells of removed pellet material and an aqueous suspension liquid.

The invention relates to the technical field of modern biomedicine, in particular to a method for promoting differentiation of human umbilical cord mesenchymal stem cells based on an EVs composite material, which comprises the following steps: 1, preparing a double-layer composite scaffold; 2, preparation and characterization of EVs: primary cartilage cell culture, enzymatic digestion of cartilage tissue, digestion termination and cell collection, EVs extraction, and EVs characterization identification; 3, loading the EVs; 4, separation, identification and differentiation induction of the hUC-MSCs; and 5, differentiation effect detection. The method has the characteristics that the differentiation efficiency and stability are good, the EVs delivery efficiency is remarkably improved, the biocompatibility and safety are good, the operation is simple and convenient, the cost is controllable, and the application scene is flexible and wide.

The invention discloses application of cornua cervi pantotrichum tissue-derived exosome-like nanoparticles in preparation of a medicine for preventing and treating senile cardiovascular diseases. A sterile, stable and high-purity cornua cervi pantotrichum tissue exosome is obtained by adopting an enzyme digestion-differential centrifugation-ultra-high-speed centrifugation combined method, the physiological efficacy of the cornua cervi pantotrichum tissue exosome in heart protection is studied, and it is found that the cornua cervi pantotrichum tissue exosome can significantly improve cardiac dysfunction of acute aging mice induced by D-galactose (D-Gal). The cardiac ejection function, the cardiac fibrosis and the inflammation invasion level are remarkably improved. The compound has treatment potential in preparation of medicines for preventing heart protection and treating heart diseases, and provides a new strategy and a treatment method for treating cardiovascular diseases of old people.

The field of this invention relates to immunohistochemistry (IHC) and in situ hybridization (ISH) for the targeted detection and mapping of biomolecules (e.g., proteins and miRNAs) in tissues or cells for example, for research use and for clinical use such by pathologists (e.g., biomarker analyses of a resected tumor or tumor biopsy). In particular, the use of mass spectrometric imaging (MSI) as a mode to detect and map the biomolecules in tissues or cells for example. More specifically, the field of this invention relates to photocleavable mass-tag reagents which are attached to probes such as antibodies and nucleic acids and used to achieve multiplex immunohistochemistry and in situ hybridization, with MSI as the mode of detection / readout. Probe types other than antibodies and nucleic acids are also covered in the field of invention, including but not limited to carbohydrate-binding proteins (e.g., lectins), receptors and ligands. Finally, the field of the invention also encompasses multi-omic MSI procedures, where MSI of photocleavable mass-tag probes is combined with other modes of MSI, such as direct label-free MSI of endogenous biomolecules from the biospecimen (e.g., tissue), whereby said biomolecules can be intact or digested (e.g., chemically digested or by enzyme).

The invention belongs to the technical field of aquatic organisms, and discloses a sycp3 molecular identification method for screening large yellow croaker oogonial stem cells and application of the sycp3 molecular identification method. The method comprises the following steps: taking out a fish ovary, and carrying out enzyme digestion and physical separation to obtain ovary; the number of the ovum is rapidly counted through AO-EB double-staining kit detection and AI software point camera artificial intelligence technology; and based on the obtained result, carrying out dual molecular identification of the stemness by means of the sycp3 gene. The sycp3 gene is used as a specific marker of the oogonial stem cells of the pseudosciaena crocea, and the sensitivity is improved by 50% or above compared with that of traditional vasa gene detection.

The present invention provides a human bone marrow organoid and its construction method and application. The construction method includes the following steps: extracting microvascular fragments from human adipose tissue by enzymatic digestion; preparing pre-vascularized cartilage tissue by dynamic culture: mechanically preparing human micro-adipocyte tissue, adding human micro-adipocyte tissue and microvascular fragments to a bioreactor in a predetermined ratio, adding culture medium for dynamic culture to obtain pre-vascularized self-aggregated adipose tissue; transplanting the pre-vascularized cartilage tissue subcutaneously into a target animal; after a predetermined period of time, the pre-vascularized cartilage tissue develops into humanized bone marrow organoids with specific functions in the target organism. The human bone marrow organoids obtained by the construction method provided by the present invention have three complete structural components: the endosteal niche, the vascular niche, and the hematopoietic cell niche, and maintain the physiological function of human hematopoietic stem cells.

The invention provides a novel nasal mucosa-derived pluripotent stem cell, a separation method and application, and belongs to the technical field of stem cell biotechnology and tissue engineering. According to the method, the technical scheme of'no enzyme digestion + continuous two-stage factor induction + three-dimensional sphere screening 'is creatively adopted, and the novel nasal mucosa-derived pluripotent stem cells are efficiently separated and enriched from nasal mucosa tissues. The method is simple to operate, reduces the risk of cell injury, can accurately screen out novel nasal mucosa-derived pluripotent stem cells expressing CD109 + and CD55 + surface markers and Nanog / Oct4 / Sox2 / Klf4 pluripotent core transcription factors, and solves the problems of low purity and instable dryness in the traditional method. Meanwhile, the nasal mucosa tissue is convenient to obtain, trauma is small, and compared with BMSCs (bone marrow mesenchymal stem cells) needing bone marrow puncture and ADMSCs (adipose mesenchymal stem cells) needing liposuction, the clinical application threshold is greatly lowered.

Methods are provided for isolating and expanding gastrointestinal (GI) tract stem cells from nonhuman primates (NHP)s. The method includes recovering a portion of GI tissue from a NHP, enzymatically digesting the tissue in a digestion medium comprising a tissue dissociation enzyme, and isolating cells through filtration and centrifugation. The isolated cells are cultured in a two-dimensional or three-dimensional (3D) culture system using an expansion medium comprising a basal formulation supplemented with one or more growth factors, vitamins, amino acids, and pathway modulators. In some embodiments, the cells are suspended in a basement membrane extract and plated as a 3D structure for organoid formation. The resulting stem cells or organoids may be analyzed using microscopy, flow cytometry, or transcriptomic profiling, and may be used for preclinical drug testing, disease modeling, or toxicity screening. The methods reduce reliance on live animal testing and improve translational relevance of in vitro gastrointestinal models.

The utility model discloses a bioreactor microcarrier adherent cell pancreatin digestion device, which belongs to the technical field of cell pancreatin digestion, and comprises a pancreatin digestion unit connected with a bioreactor, and the pancreatin digestion unit is connected with a bottom valve of the bioreactor through a second connecting pipe; the micro-carrier adherent cells are digested by pancreatin in the bioreactor. The bioreactor microcarrier adherent cell pancreatin digestion device provided by the utility model can be used for observing the cell digestion state in real time and terminating digestion in time, carrying out data statistics and analysis on cell counting, calculating the recovery rate of digested cells, and observing the cell attachment rate and the cell growth condition of the cells in a next-stage bioreactor, so that the digestion efficiency of the cells in the next-stage bioreactor is improved. Compared with culture parameter data of a production batch for cell digestion by using a butterfly-shaped pancreatin digester before improvement, the recovery rate and the adherence rate are stably increased, and the working efficiency and the product yield are better improved.

The present disclosure provides, for example, double stranded DNA (dsDNA) molecules comprising a chemically modified uridine nucleotide. In some embodiments, the dsDNA molecules comprise a therapeutic load sequence. In some embodiments, the dsDNA molecules are resistant to endonuclease digestion and / or to immunosensor recognition, and support the expression of a therapeutic load encoded in the dsDNA molecules. The present disclosure also provides, for example, pharmaceutical compositions comprising dsDNA molecules comprising a chemically modified uridine nucleotide.

The invention provides a method for separating and enriching rabbit carotid artery stem cells, and belongs to the technical field of animal cell culture. The method specifically comprises the following steps: putting a rabbit carotid artery body of which surface blood and impurities are removed into a culture plate containing a carotid artery body stem cell culture medium for adherent culture, and inducing cells to climb out; and removing rabbit carotid artery tissues, carrying out digestion passage on the crawl-out cells, carrying out suspension culture by using a carotid artery stem cell culture medium, and inducing the cells to form balls. The invention provides a method based on combination of tissue adherent culture and suspension balling. The method is used for efficiently separating and enriching stem cells from adult rabbit carotid body tissues and inducing to form stem cell spheres. Meanwhile, the method provided by the invention does not need enzymatic digestion, is high in balling efficiency, is suitable for obtaining the carotid stem cells of the rabbit model, and has definite innovativeness and wide application prospects.

Previously, seasonings made from sake lees have been known to enhance umami and richness, as well as reduce the odors of meat and seafood. However, there is no knowledge about its effect in making meat and seafood feel moist and tender. [Solution] We offer a seasoning that uses digested sake lees (sake lees) as an ingredient, obtained by digesting (breaking down) sake lees with koji and enzymes, resulting in a moist and tender texture for meat and seafood.

A low-input RNase footprinting approach is described for the rapid quantification of ribosome-protected fragments with 1,000-100,000 cells. The assay uses a simplified procedure for capturing ribosome-RNA complexes based on optimized RNase digestion. It simultaneously maps cytosolic and mitochondrial translation with single-nucleotide resolution.

The invention provides an IGH rearrangement detection reverse transcription primer group, a quantitative screening detection kit and application. The reverse transcription primer group is specifically designed aiming at common IGH breaking sites, and a T basic group close to the 3'end in a reverse transcription primer is not modified, so that the normal function of reverse transcriptase is not influenced; a T basic group at the 5'end of the reverse transcription primer is replaced by a U basic group, so that the reverse transcription primer can be digested by uracil-N-glycosylase (UNG); after reverse transcription is finished, residual reverse transcription primers can be partially digested by UDG, reverse transcription is performed through a reverse transcription primer group to obtain IGH gene cDNA and different types of fusion gene cDNA, and quantitative detection of any one of the three types of rearrangement can be realized by combining specific primers and probes designed at the upstream of DUX4, EPOR, CRLF2 and ABL1 genes respectively. The kit has significant application value in clinical auxiliary diagnosis.

The present invention relates to a method for ethanol-free messenger RNA (mRNA) purification, and specifically provides a method for purifying mRNA by subjecting a preparation comprising in vitro synthesized mRNA to one or more steps of enzymatic digestion with a proteolytic enzyme, and optionally further steps of low dT affinity chromatography. Also provided are mRNA purified by the methods of the invention.

The application discloses a sequencing library for removing human-derived nucleic acid and a construction method and application thereof, and the construction method comprises the following steps: fragmentizing nucleic acid and adding sequencing adapters, constructing a DNA or RNA library, or constructing a RNA and DNA co-library, then taking the library with the added sequencing adapters as a template, performing denaturation annealing on the library and specific human-derived hybridization probes, and performing digestion on the library by using DSN enzyme, and in the renaturation process, human-derived nucleic acid with a higher content is preferentially annealed with the human-derived hybridization probes to form double strands, which are degraded by the DSN enzyme, pathogenic microorganism with a low content is slowly renatured and remains in a single-stranded state, and cannot be digested by the DSN enzyme, and the human-derived nucleic acid is effectively removed, and the nucleic acid of the pathogenic microorganism is further enriched through subsequent PCR. The method can effectively reduce the proportion of human-derived nucleic acid in the library, and maximize the retention of nucleic acid information of the pathogenic microorganism.