230 results about "Abnormal alkaline phosphatase" patented technology

The invention belongs to the technical field of biological medicine, and discloses a probiotic agent capable of improving the physiological activity function of vitamin D and maintaining bone health and application of the probiotic agent. The invention provides a probiotic agent composed of bifidobacterium adolescentis and bifidobacterium longum, the probiotic agent can significantly improve the bone mineral density, the bone trabecula thickness, the bone trabecula number, the bone trabecula connectivity and the bone volume fraction of an osteoporosis mouse, and meanwhile, the bone trabecula separation degree of the osteoporosis-removed mouse is significantly reduced; the level of vitamin D metabolites such as 25-hydroxy vitamin D and 1, 25-dihydroxy vitamin D in mouse serum is improved; the content of serum calcium and alkaline phosphatase in serum of mice is reduced, and the level of osteocalcin and type I procollagen amino terminal peptide in the serum is improved. The probiotic agent can relieve bone calcium loss, maintain bone health, repair bone injury, increase osteoblasts and reduce the osteoporosis degree, and can be used for preparing drugs for preventing and / or treating osteoporosis or preparing health care products or food beneficial to improving bone mineral density.

The invention relates to a method for improving chemiluminescence immunoassay sensitivity of magnetic particles, a detection kit and application thereof, firstly, magnetic beads are coupled with an antibody, then magnetic beads are sealed to obtain a magnetic bead coated antibody, sealing comprises pre-sealing and second-step sealing, glycine and ethanolamine are added in the pre-sealing process, and in the second-step sealing process, glycine and ethanolamine are added in the second-step sealing process. Casein hydrolysate and dodecyl dimethyl amine ethyl lactone are added; then coupling the antibody with alkaline phosphatase or streptavidin to obtain an alkaline phosphatase labeled antibody or a streptavidin labeled antibody; the magnetic bead coated antibody, the alkaline phosphatase labeled antibody or the streptavidin labeled antibody is used for sample detection. The method is high in sensitivity and wide in detection range, is adaptive to a common immunoassay instrument, does not need expensive equipment, and is also simple to operate; in addition, the AMH concentration with extremely low concentration can be accurately detected.

The invention belongs to the technical field of preparation of polypeptides, and particularly relates to a self-assembled polypeptide capable of responding to alkaline phosphatase and degrading Neuropilin-1 as well as a preparation method and application of the self-assembled polypeptide. The self-assembled polypeptide (formula I) provided by the invention has a self-assembly starting unit 'GNNQQNY (SEQ ID NO: 1)', can be self-assembled in vitro, and can form nanofibers after responding to alkaline phosphatase, so that not only can the critical self-assembly concentration of the polypeptide be reduced, but also the binding capacity with target protein can be improved. The material can degrade NRP1 on the surfaces of tumor cells and Treg cells, and has huge potential in promoting immunotherapy of tumors.

The invention relates to the field of biomedicine, particularly provides a copper-based nano-enzyme as well as a preparation method and application thereof, and aims to solve the problems that in the prior art, clinical treatment on primary sclerosing cholangitis (PSC) is difficult in diagnosis and lacks of effective treatment drugs. The copper-based nano-enzyme comprises a nano-enzyme carrier and a copper-based nano-enzyme, wherein the nano-enzyme carrier is a two-dimensional nanosheet formed by copper ions, gallic acid and ursodesoxycholic acid through coordinate bonds; the probe molecule is a cyanine dye molecule connected to the surface of the nano-enzyme carrier through a covalent bond; wherein the fluorescence intensity of the copper-based nano enzyme is enhanced after the action of the alkaline phosphatase. The three functions of alkaline phosphatase responsive diagnosis, nano-enzyme catalytic treatment and ursodesoxycholic acid hepatic targeting are innovatively and synergistically integrated into one nano platform, the treatment function can be executed while specific imaging diagnosis is performed on diseases, and a new strategy is provided for diagnosis and treatment of diseases such as PSC.

The invention discloses an antibody for detecting indol sulfate, a detection kit and application of the antibody, and the antibody can be specifically combined with indol sulfate and has high combination activity. The detection kit detects the content of indoxyl sulfate in a to-be-detected sample through a competitive binding reaction of an immunomagnetic bead coated indoxyl sulfate antibody, an antigen in the sample and an alkaline phosphatase labeled indoxyl sulfate antigen, and has important economic value and social significance.

Featured are polypeptides that include soluble alkaline phosphatases, mutants, fragments, fusion proteins thereof, and methods of use thereof, for treating bone mineralization disorders, such as hypophosphatasia (HPP), and symptoms thereof. The polypeptides include a soluble alkaline phosphatase (sALP) or fragment thereof, which is derived from a naturally occurring alkaline phosphatase (ALP).

The invention discloses an eggshell membrane peptide for enhancing bone mineral density as well as a preparation method and application thereof, belongs to the technical field of egg by-product processing, and particularly relates to a preparation method of the eggshell membrane peptide for enhancing bone mineral density, which comprises the following steps: carrying out vacuum freeze drying and crushing on an eggshell membrane to obtain eggshell membrane powder; mixing the eggshell membrane powder with deionized water to obtain an eggshell membrane solution; adding keratinase, performing enzymolysis, and taking supernate; adding collagenase, carrying out enzymolysis, and centrifuging to take supernate, so as to obtain primary enzymatic hydrolysate; adding alkaline phosphatase, performing enzymolysis and centrifugation, and taking supernate to obtain enzymatic hydrolysate; and performing ultrafiltration and drying to obtain the eggshell membrane peptide. According to the eggshell membrane peptide for enhancing the bone mineral density, keratin cross-linking is destroyed through enzymolysis of keratinase, active peptide capable of stimulating osteoblast proliferation is released through enzymolysis of collagenase, phosphopeptide calcium binding sites are further activated through enzymolysis of alkaline phosphatase, calcium salt deposition is promoted, and the eggshell membrane peptide for enhancing the bone mineral density is provided.

The invention discloses a nanofiber based on a polypeptide derivative and an enzymatic self-assembly method and application of the nanofiber based on the polypeptide derivative, the sequence of the polypeptide derivative is Nap-Gly-Phe-Phe-pTyr-Lys-Trp-Tyr-Gln-Asn-Met-Ile-Arg, and the Gly-Phe-Phe-pTyr is of an L configuration or a D configuration. The self-assembled polypeptide derivative disclosed by the invention contains an alkaline phosphatase restriction enzyme cutting site, and the self-assembled polypeptide derivative is self-assembled at a specific part with high enzyme specificity expression through enzyme catalysis to form nanofibers. The nanofiber hydrogel obtained through enzymatic treatment is carried out under physiological conditions, high temperature or ultraviolet irradiation is not needed, damage of high temperature to bioactive substances is avoided, the nanofiber hydrogel is more suitable for a temperature-sensitive in-vivo environment, toxic chemical cross-linking agents or photoinitiators do not need to be introduced, the cytotoxicity of the material is remarkably reduced, active enzymes or drugs can be embedded in situ, and the nanofiber hydrogel can be used for in-situ treatment. The biological function is maintained.

The invention discloses a probe set, a kit and a nucleic acid signal amplification in-situ hybridization detection method, and relates to the technical field of molecular biology, the probe set comprises: a labeled extension probe, the sequence of which is as shown in SEQ ID No.9-24; and the sequence of the closed probe is as shown in SEQ ID No. 25-27. The kit comprises the probe set, a signal amplification precursor, a signal amplification body and an alkaline phosphatase labeled probe. According to the present invention, the partial probe sequence introduces the non-natural bases, and can be used for detecting the HPV18 infected cervical cancer Hela cells, the experiment results show that the introduction of the non-natural bases does not affect the normal positive value, the matching degree between the non-natural bases is more specific than the matching of the natural bases, and the detection sensitivity is improved.

The present invention discloses a type I semi-cyanine photosensitizer, a type I targeted photosensitizer, and a preparation method and application thereof. The semi-cyanine photosensitizer can rapidly generate a large amount of O2 under light and hypoxic conditions. ·‑ , thereby achieving the effect of photodynamic therapy, and having good biocompatibility and biodegradability, it is an effective type I photosensitizer. In addition, the above-mentioned type I photosensitizer is reacted with phosphorus oxychloride in the presence of water to prepare a type I photosensitizer with tumor targeting. It can specifically target alkaline phosphatase, a biomarker highly expressed in tumors. Under the action of alkaline phosphatase, the photosensitizer can be enriched in the tumor area, thereby achieving precise tumor imaging and photodynamic therapy, which can effectively reduce the side effects of the photosensitizer and improve the inhibitory effect of the photosensitizer on tumors.

The invention provides a multicolor visual semi-quantitative detection method for miRNA in skin interstitial fluid, which comprises the following steps: extracting a skin interstitial fluid sample in a minimally invasive manner by combining a microneedle patch with a vacuum negative pressure technology; the method comprises the following steps: incubating a skin interstitial fluid sample and a magnetic nanoprobe of which the surface is immobilized with a hairpin probe H1, opening H1 in the presence of target miRNA, triggering a subsequent hybridization chain reaction of a hairpin probe H2 and H3, and constructing a biotin-rich DNA compound on the surface of a magnetic bead; the method comprises the following steps: carrying out enzyme labeling by using an alkaline phosphatase-streptavidin conjugate, catalyzing ascorbic acid phosphate to generate ascorbic acid to reduce potassium iodate, and generating an iodine elementary substance to etch a gold nano bipyramid, so that the color of a solution is changed, and high-sensitivity detection on target miRNA is realized. The multi-color visual detection method provided by the invention is combined with the gold nano bipyramid localized surface plasmon resonance property change, so that near-full spectrum color change can be generated, and the portability of the sensor and the accuracy of visual analysis can be improved.

The invention provides an alkaline phosphatase color developing solution and a preparation method thereof, and particularly relates to the technical field of immunodetection, the alkaline phosphatase color developing solution is formed by dissolving chromogen dry powder in a color developing buffer solution, and the concentration is 1-3 mg / ml; wherein the chromogen dry powder is composed of naphthol AS-MX phosphate and fast red-TR; the preparation method comprises the following steps: S1, dissolving naphthol AS-MX phosphate and fast red TR with purified water to obtain a solution; wherein the concentration of naphthol AS-MX phosphate in the solution is 0.5 to 1.5 mg / ml, and the concentration of fast red TR is 0.5 to 1.5 mg / ml; s2, freezing and storing the solution in a refrigerator, and condensing the solution into ice blocks; s3, the ice blocks are put into a freeze dryer, vacuumizing is carried out, water molecules in the ice blocks are sublimated into water vapor under the negative pressure condition, the water vapor is discharged, and chromogen dry powder can be obtained; s4, the chromogen dry powder is dissolved in the chromogenic buffer solution, and an alkaline phosphatase chromogenic solution can be obtained; the prepared developing solution has the core advantages of high sensitivity, rapid developing, multicolor compatibility, digital adaptation and the like, and an efficient and reliable tool is provided for pathological research and clinical diagnosis.

The invention provides application of linariin-6-O-glucoside in preparation of a product for preventing and treating osteoporosis, and relates to the field of biological medicine. According to the invention, dyeing experiments of alkaline phosphatase, alizarin red and tartaric acid resistant acid phosphatase prove that the sesaminin-6-O-glucoside has affinity for FPPS receptors, has the capability of promoting bone marrow mesenchymal stem cells to form bone cells, is superior to that of bisphosphate, and can inhibit the generation of osteoclasts; microCT (micro computed tomography) detection results show that the sesaminin-6-O-glucoside has the effect of relieving the osteoporosis and can promote the formation of bone trabecula. According to the application disclosed by the invention, it is proved for the first time that the sesaminin-6-O-glucoside has the effect of resisting osteoclast formation, and meanwhile, the sesaminin-6-O-glucoside also has a better osteogenesis effect, so that a basis is provided for developing the sesaminin-6-O-glucoside as a potential medicine for treating osteoporosis.

The invention discloses a cyclosporine detection kit and application thereof, the cyclosporine detection kit comprises a reagent R1 comprising a magnetic bead coated cyclosporine antibody and a Tris buffer solution I, a reagent R2 comprising an alkaline phosphatase labeled cyclosporine antigen and an AP stabilizer, a reagent R3 comprising an MES buffer solution, and a reagent R4 comprising an MES buffer solution. The sample treating agent comprises a Tris buffer solution III containing copper sulfate pentahydrate, sodium sulfite, a preservative, methanol and ethylene glycol. According to the cyclosporin detection kit and the application thereof, the R1, R2 and R3 reagents are combined and applied to the to-be-detected sample treated by the specific Tris buffer solution I sample treating agent, the specific detection of cyclosporin is improved through the synergistic effect of the specific Tris buffer solution I, the AP stabilizer and the specific MES buffer solution, the detection accuracy and stability are good, the repeatability is good, and the kit is suitable for large-scale popularization and application. The problems of cross reaction and insufficient stability during detection of a to-be-detected sample are solved.

The present application relates to the technical field of biotechnology, and particularly relates to a D-dimer chemiluminescence assay kit. The present application provides a D-dimer chemiluminescence assay kit, which comprises a magnetic particle suspension (immunomagnetic beads) coated with D-dimer antibody 1, alkaline phosphatase-labeled D-dimer antibody 2 (enzyme-labeled antibody) and a calibrant. The immunomagnetic beads and the enzyme-labeled antibody are prepared by chemical coupling and stored in a buffer containing a soluble high molecular material, which can effectively improve the detection sensitivity and repeatability. The kit can be used in combination with a substrate solution containing a chromogenic substrate such as adamantane and its derivative AMPPD or APS-5 for a full-automatic immune test system, and the content of D-dimer in plasma or whole blood can be determined by a chemiluminescence instrument. The kit has high detection sensitivity and good repeatability.

Featured are polypeptides that include soluble alkaline phosphatases, mutants, fragments, fusion proteins thereof, and methods of use thereof, for treating bone mineralization disorders, such as hypophosphatasia (HPP), and symptoms thereof. The polypeptides include a soluble alkaline phosphatase (sALP) or fragment thereof, which is derived from a naturally occurring alkaline phosphatase (ALP).

The invention provides acellular matrix hydrogel loaded with animal acellular omentum particles as well as a preparation method and application of the acellular matrix hydrogel loaded with the animal acellular omentum particles, and relates to the technical field of tissue engineering and regenerative medicine. Comprising a decellularization treatment agent, a decellularization matrix, functional particles, an enzymolysis agent, a cross-linking stabilizer, a functional additive and a solvent, the functional additive is prepared from PDA coated Fe3O4 magnetic nano particles, phosphorylated polyglycerol sebacate and liposome coated silver nano particles. The PDA-Fe3O4 magnetic nanoparticles of the functional additive generate mechanical signals under the action of an external magnetic field, a Piezo1 mechanical gating cation channel is activated to promote calcium ion internal flow, and the autonomous mineralization ability of phosphorylated polyglycerol sebacate is combined, so that the activity of the bone differentiation marker alkaline phosphatase is effectively improved, and the bone differentiation marker alkaline phosphatase is prepared. The combination of the PDA coated Fe3O4 magnetic nanoparticles and the phosphorylated polyglycerol sebacate integrates a magnetic response mineralization system.

Bispecific antibody constructs comprising a first binding domain, which binds to human alkaline phosphatase, placental type (ALPP) and / or alkaline phosphatase, germ cell type (ALPPL2), on the surface of a tumor cell and a second binding domain, which binds to human CD3 on the surface of a T cell.

The invention relates to a calcitonin luminescence detection kit and a detection method and application thereof. Specifically, the invention provides the high-sensitivity calcitonin luminescence detection kit, the high-sensitivity calcitonin luminescence detection kit comprises a first reagent and a second reagent, and the first reagent comprises magnetic beads connected with first protein; the second reagent comprises alkaline phosphatase connected with the second protein. The kit improves the detection accuracy and the sample coincidence rate.

The invention provides a preparation method of an alpha-2, 3 sialic acid glycosyl transferase mediated sugar chain fixed-point coupling antibody drug conjugate, which comprises the following steps: S1, dissolving an antibody, adding UDP-Gal, Mn < 2 + > and beta-1, 4 galactosyl transferase for reaction, and purifying after reaction to obtain an antibody I with galactose at the tail end of a sugar chain; s2, dissolving the antibody I, adding an azide-modified CMP sialic acid derivative, alpha-2, 3 sialic acid glycosyltransferase and alkaline phosphatase for reaction, and purifying after reaction to obtain an antibody II of which the tail end of a carbohydrate chain is connected with the sialic acid derivative; s3, dissolving toxin molecules in DMSO, adding the dissolved toxin molecules into the antibody II for reaction, and performing ultrafiltration after the reaction to obtain ADC molecules; or the ADC molecule is directly prepared by adopting a one-pot enzyme method. The screened ST3Gal3 is used for preparing the carbohydrate chain fixed-point coupling ADC, the DAR value is controllable, batch-to-batch stability is achieved, uniformity is high, repeatability is high, quality control and CMC are better, and then the drug effect of the ADC can be better improved.

The invention discloses a preparation method of pichia pastoris expression secreting type deglycosylation alkaline phosphatase, and belongs to the technical field of biological engineering. According to the invention, the GS115-BIAP II pichia pastoris genetically engineered bacterium capable of secreting and expressing bovine intestine alkaline phosphatase BIAP II is prepared by using a conventional method. The method comprises the following steps: carrying out site-directed knockout on an ALG3 gene for coding alpha-1, 3-mannosyl transferase and an OCH1 gene for coding alpha-1, 6-mannosyl transferase by using a CRISPR-Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats-associated Protein 9) technology to prepare a glycosylation gene defect type engineering bacterium GS115-delta ALG3-delta OCH1-BIAP II. The secretory expression BIAP II is deglycosylated alkaline phosphatase, and the specific activity of the secretory expression BIAP II is close to that of natural BIAP II. The deglycosylated alkaline phosphatase can improve the antibody coupling efficiency and marking uniformity under specific conditions. The method has remarkable advantages in chemiluminescence diagnosis, high-sensitivity immunodetection and development of multifunctional biological coupling reagents.

The invention relates to an anti-ovarian antibody chemiluminiscence kit and a preparation method thereof. The kit comprises amino magnetic beads coated with anti-ovarian antibody recombinant protein, anti-human immune globulin labeled by alkaline phosphatase, an anti-ovarian antibody calibrator and a chemiluminiscence substrate. The method has the beneficial effects that the 7-day acceleration stability of the AOAb coated amino magnetic bead is improved to 95% or above, the signal-to-noise ratio is high, the correlation coefficient is greater than 0.999, the test repeatability CV is less than 3.5%, the reagent correlation and the magnetic bead stability can be better improved, and the test result is ensured to be accurate. By optimizing the amino magnetic bead cross-linking agent, 60 clinical samples are tested and compared with the result of a comparison manufacturer, the total coincidence rate is as high as 98%, and false positive can be effectively reduced. Meanwhile, a detection system is formed by matching with a Guilin superior chemiluminescence immunoassay analyzer IA-260, the testing amount per hour reaches 130 tests, and compared with a traditional colloidal gold immunochromatography method and a traditional enzyme-linked immunosorbent assay method, the detection speed is greatly increased, and the result optimization accuracy is greatly improved.

This invention relates to the field of chemiluminescence detection technology for in vitro diagnostic reagents, specifically to an S100-β chemiluminescence detection kit and its preparation method. It provides a chemiluminescence detection kit for central nervous system-specific proteins using magnetic microparticles. The kit comprises: S100-β calibrator, S100-β monoclonal antibody conjugated with carboxyl magnetic beads, S100-β monoclonal antibody labeled with alkaline phosphatase, AMPPD chemiluminescence substrate solution acted upon by alkaline phosphatase, and concentrated washing buffer. This kit achieves the following: no sample pretreatment required, easy operation, high automation, high sensitivity, automatic result output in 16 minutes, good stability of calibrator and enzyme label, and low cost. It can meet the clinical needs for auxiliary diagnosis and treatment of acute ischemic brain injury.

Provided are a method for quantifying an amount of capped messenger RNA (mRNA) in an mRNA sample comprising contacting the mRNA with two or more of a nuclease, an alkaline phosphatase, and a polynucleotide kinase and separating the capped mRNA and the uncapped mRNA occurs using chromatography.

The present invention discloses a split-type photoelectrochemical biosensing analysis method based on a Bi2S3 / Ag3PO4 heterojunction composite material. First, a Bi2S3 / Ag3PO4 heterojunction is prepared by multi-step hydrothermal method, and then modified on the surface of a fluorine-doped tin oxide electrode to form a Bi2S3 / Ag3PO4 photoelectrode; when a target is present, the capture aptamer on the surface of the magnetic bead can capture the triggering aptamer chain, thereby triggering the HCR reaction. With the help of the specific recognition effect between streptavidin and biotin, alkaline phosphatase is introduced on the surface of the magnetic bead to catalyze the substrate to form ascorbic acid as an electron donor, thereby enhancing the current. Carcinoembryonic antigen is quantitatively analyzed by photocurrent signal. This analysis method combines hybridization chain reaction and enzymatic signal amplification strategy, and has the advantages of simple preparation, high sensitivity, and reusability.

The present disclosure features methods for treating neutralizing antibodies that reduce the efficacy of soluble alkaline phosphatase therapy (e.g., asfoenzyme alpha), which is an enzyme replacement therapy, such as for the treatment of bone mineralization disorders, e.g., hypophosphoesterase (HPP). The methods include diagnosing the subject for the presence of a neutralizing antibody that affects the efficacy of the treatment, administering a treatment suitable for reducing the deleterious effects of the neutralizing antibody, and continuing an alkaline phosphatase treatment.

The invention provides a composite growth factor composition capable of promoting proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, a method and application. The invention provides the composite growth factor composition by simulating a natural secretion mode of a decalcified bone matrix of the mandible of an SD (Sprague Dawley) rat on the seventh day, the composite growth factor composition comprises BMP-2 (400-600 pg / ml), TGF-beta1 (5-7 ng / ml) and IGF-1 (150-250 ng / ml), and the optimal concentrations are 500 pg / ml, 6 ng / ml and 200 ng / ml respectively. The composition is remarkably superior to a single factor and a high-dosage combination in the aspects of promoting BMSCs proliferation, improving the activity of alkaline phosphatase (ALP) and forming mineralized nodules, meanwhile, the use dosage of growth factors is remarkably reduced, and the risk of side effects such as heterotopic ossification is reduced. The composite growth factor composition has good biocompatibility and safety, and is suitable for the fields of bone tissue engineering, bone defect repair and related regenerative medicine.

This application relates to targeting peptides and engineering AAV capsids. In some embodiments, the targeting peptides and engineered AAV capsids are capable of interacting with ALPL (Tissue Non-Specific Alkaline Phosphatase) to mediate delivery to a cell or tissue.

The invention relates to a gold nanoparticle probe modified with a fluorescent molecule responsive to alkaline phosphatase and a preparation method thereof, belonging to the field of probe technology. The probe is hCy‑ALP@AuNPs, and the composite structure is mainly composed of gold nanoparticles of about 20 nm, and the surface is modified with a fluorescent molecule responsive to alkaline phosphatase through a bioorthogonal reaction of thiol-polyethylene glycol-diphenylcyclooctyne. The fluorescent molecule consists of a recognition group of alkaline phosphatase and a near-infrared fluorescent dye IR700. hCy‑ALP@AuNPs can achieve the effect of fluorescence onset under the hydrolysis of the tumor marker alkaline phosphatase. Compared with a single fluorescent probe, the grafted gold nanoparticles of the present invention have a high retention (EPR) effect in solid tumors, which can enhance their accumulation effect in the tumor site, increase the imaging window time, and achieve the purpose of long-retention imaging. The present invention has important application value in the fields of drug delivery carriers, bioimaging, medicine, etc.

The application discloses a kind of self-assembly nanomaterial and its application in ROS clearance.The self-assembly nanomaterial of the application is denoted as TPP-FFYp-O, and its structural formula is shown as follows:TPP-FFYp-O of the application is specifically cut to generate TPP-FFY-O by alkaline phosphatase (ALP) overexpressed after entering inflammatory cell, TPP-FFY-O is enriched near mitochondrion by the targeting effect of TPP, and is self-assembled to form nanoparticle and stays in inflammatory cell by π-π stacking.Nanoparticle surface enrichment of a large number of TEMPO can convert superoxide radical anion (O2 ·‑ ) in mitochondrion into hydrogen peroxide (H2O2), and then decompose into oxygen and water, thereby significantly reducing the ROS level in inflammatory cell.This enzyme-controlled self-assembly nanoparticle improves the stability of TEMPO and the ability of ROS clearance, and has good application prospect.