51 results about "Arylsulfatase" patented technology

The present invention pertains to a process for production of recombinant arylsulfatase A in a cell culture system, the process comprising culturing a mammalian cell capable of producing rASA in liquid medium in a system comprising one or more bio-reactors; and concentrating, purifying and formulating the rASA by a purification process comprising one or more steps of chromatography. Other aspects of the invention provides a pharmaceutical composition comprising rASA, which is efficiently endocytosed via the mannose-6-phosphate receptor pathway in vivo as well as a rhASA a medicament and use of a rhASA for the manufacture of a medicament for reducing the galactosyl sulphatide levels within target cells in the peripheral nervous system and / or within the central nervous system in a subject. A final aspect of the invention provides a method of treating a subject in need thereof, said method comprising administering to said subject a pharmaceutical composition comprising a rhASA and thereby obtaining a reduction in the galactosyl sulphatide levels in target cells within said subject.

Provided herein are methods and compositions for treating a subject suffering from a deficiency in arylsulfatase A in the CNS. The methods include systemic administration of a bifunctional fusion antibody comprising an antibody to a human insulin receptor and an arylsulfatase A.

Provided herein are methods and compositions for treating a subject suffering from a deficiency in arylsulfatase A in the CNS. The methods include systemic administration of a bifunctional fusion antibody comprising an antibody to a human insulin receptor and an arylsulfatase A.

The present invention provides, among other things, compositions and methods for CNS delivery of lysosomal enzymes for effective treatment of lysosomal storage diseases. In some embodiments, the present invention includes a stable formulation for direct CNS intrathecal administration comprising an arylsulfatase A (ASA) protein, salt, and a polysorbate surfactant for the treatment of Metachromatic Leukodystrophy Disease.

A purified nucleic acid molecule capable of expressing a lysosomal enzyme, e.g., iduronidase or arylsulphatase, is provided. The molecule includes a promoter of the phosphoglycerate kinase gene to facilitate expression of the lysosomal enzyme in the brain. Expressing the lysosomal enzyme in the brain of patients with lysosomal storage diseases, such as mucopolysaccharidosis, provides treatment for the neurological aspects of these diseases. The invention also provides a method of gene therapy for lysosomal storage diseases by administering the nucleic acid molecule in the form of a vector.

A method for the stable expression of an introduced exogenous gene in a plant or plant cell is provided. Stable expression of an exogenous gene that was introduced was achieved by operably linking an upstream sequence of sea urchin arylsulfatase gene as an insulator.

The invention relates to a strain for producing aryl sulfatase and application thereof, and particularly provides a strain with preservation number of CCTCC NO: M 2013613, application of the strain in production of aryl sulfatase, a method for producing the aryl sulfatase by use of the strain and a method for producing agar with high gel strength and low electroosmosis. The screened strain can effectively produce the aryl sulfatase which can be used for carrying out enzymatic hydrolysis on agar powder and specifically removing sulfate radicals of agaropectin polysaccharide, and the yield of the agar with high gel strength and low electroosmosis can be greatly improved, and therefore, the strain has important application value in industrial production.

The invention belongs to the technical field of organic pollutant analysis, and discloses a solid-phase extraction-liquid chromatography triple quadrupole mass spectrometry isotope dilution method foronline determination of hydroxyl polycyclic aromatic hydrocarbon in urine. The method comprises the following steps: adding an isotope internal standard into urine, and then adding beta-glucuronidase-aryl sulfatase; carrying out enzymolysis on a constant-temperature shaking table, adding an organic solvent to precipitate protein after enzymolysis, carrying out refrigerated centrifugation, transferring the centrifuged supernatant, carrying out online solid-phase extraction through automatic valve switching, and then carrying out quantitative determination through liquid chromatography triple quadrupole mass spectrometry; and carrying out data processing and quantitative calculation to obtain the content of the hydroxyl polycyclic aromatic hydrocarbon in the urine. According to the invention, rapid and accurate detection of hydroxyl polycyclic aromatic hydrocarbon metabolites in human urine is realized, and an effective method is provided for human exposure health risk assessment of polycyclic aromatic hydrocarbon.

An arylsulfatase and a preparation method and applications thereof, relate to an arylsulfatase with a new gene sequence and preparation thereof. Provided is arylsulfatase Ary423 originated from the Flammeovirga Pacifica H2. Preparation methods of arylsulfatase gene Ary423 and arylsulfatase Ary423 are also provided. Gene Ary423 is cloned, recombinant plasmid carrying with the gene Ary423 is constructed and then converted to the escherichia coli BL21, the positive clones are selected and cultured in a culture medium, then the cultured positive clones are re-suspended in a lysis buffer, after the processes of cracking and centrifugation, the supernatant is collected and homogeneously mixed and purified with Ni-NTA Agarose. Arylsulfatase Ary423 can stably express in escherichia coli, recombinase maintains high catalysis efficiency in a relatively wide temperature range and pH range, so that the shortages of low vigor of arylsulfatase and tedious processes of separating and purifying in the prior art are overcome.

The invention belongs to the technical field of organic pollutant analysis, and discloses a method for on-line determination of phthalate metabolite content in urine and application, the method comprises the following steps: adding an isotope internal standard into the urine, then adding beta-glucuronidase-aryl sulfatase, carrying out enzymolysis on a constant-temperature shaking table, adding an organic solvent to precipitate protein after enzymolysis, freezing and centrifuging, transferring the centrifuged supernate, switching through an automatic valve to carry out online solid-phase extraction, and then carrying out quantitative determination through liquid chromatography and triple quadrupole mass spectrometry; and carrying out data processing and quantitative calculation to obtain the content of the phthalate metabolite in the urine. According to the invention, the rapid (20-28min) accurate detection of the phthalate metabolite in the urine of the human body is realized, and an effective method means is provided for human body exposure health risk assessment of phthalate.

The invention discloses a method for measuring residual quantities of growth promoting agents such as zilpaterol, cimbuterol, clenproperol and bambuterol in beef. The method includes hydrolyzing beef samples by the aid of beta-glucuronidase / aryl sulfatase; centrifuging the beef samples under the condition of pH (potential of hydrogen) of 1.0+ / -0.2; regulating pH of supernatant until the pH reaches 9.5+ / -0.2; sequentially extracting the supernatant by the aid of ethyl acetate and tert-butyl methyl ether; combining organic phases with one another; drying the organic phases by means of blowing; dissolving the organic phases by the aid of formic acid; enabling the organic phases to pass cation exchange solid-phase extraction columns; sequentially leaching the organic phases by the aid of 2% formic acid, water and methanol; eluting the organic phases by the aid of ammonium methanol; drying eluent by means of blowing; dissolving the organic phases by the aid of formic acid and methanol aqueous solution; filtering the organic phases by the aid of films; feeding filter liquid in liquid chromatograph / mass spectrometers to obtain the residual quantities of the growth promoting agents in the beef according to standard curves. The method has the advantages that the four growth promoting agents including the zilpaterol, the cimbuterol, the clenproperol and the bambuterol can be quickly detected by the aid of the method, and residues of the four growth promoting agents which are four types of medicine in the beef can be monitored.

The invention belongs to the field of medicine, and discloses an aryl sulfatase and glucuronidase inhibitor, a preparation method thereof and application of the inhibitor. The preparation method for the aryl sulfatase and glucuronidase inhibitor comprises the following steps: adding NO or a chemical reactant capable of producing the NO by a reaction into a solvent to obtain a solution, namely thearyl sulfatase and glucuronidase inhibitor. Compared with conventional medical treatment inhibitors STX64, D-Saccharic acid,1,4-lactone and the like, the aryl sulfatase and glucuronidase inhibitor hasthe advantages of low effective inhibition concentration, good effect, simple operation, low cost, rapidness and the like, can perform inhibition of different degrees as needed, and has a wide application prospect in the medical field.

The invention relates to a Rhodococcus and its application in degrading lignin, belonging to the technical field of microorganisms; the strain is obtained by separating and purifying the sea mud of the South China Sea, and is a variant of the genus Rhodococcus named (Rhodococcussp. ) L17, which was identified as Gram-positive, non-conidia-forming, aerobic, catalase-positive, and arylsulfatase-negative; and its 16S rDNA sequence has been submitted to the NCBI database with accession number KM215139 , and was preserved on March 20, 2014 in the General Microorganism Center of China Microbiological Culture Collection Management Committee, the preservation number is CGMCC8943; it has been verified by experiments that the strain has the ability to degrade lignin, and the degradation rate can reach 90%‑100% , which can be applied in the field of environmental protection, especially in the biodegradation of lignin.

The invention belongs to the field of medicine, and discloses an arylsulfatase and beta-glucuronidase inhibitor as well as a preparation method and application thereof. The preparation method of the arylsulfatase and beta-glucuronidase inhibitor comprises the following steps of adding peracetic acid into a solvent to obtain a solution, namely the arylsulfatase and beta-glucuronidase inhibitor. Compared with conventional medical treatment inhibitors such as STX64 and D-Sacchuric acid, 1, 4-lactone and the like, the arylsulfatase and beta-glucuronidase inhibitor has the advantages of being goodin effective inhibition concentration effect, capable of achieving inhibition of different degrees according to needs, easy to operate, low in cost, rapid and the like, and has a certain application prospect in the medical field.

The present invention belongs to the field of biomedical technologies and discloses the use of arylsulfatase K in the preparation of drugs for preventing, delaying or treating chronic obstructive pulmonary disease. The present invention has for the first time discovered the role of ARSK (arylsulfatase K) in inhibiting mitophagy and delaying the senescence of airway epithelial cells, and further determined the key role of ARSK in the occurrence and development of COPD, which can provide a new target for the prevention and treatment of COPD. The present invention has determined that ARSK can slow down the process of lung senescence by alleviating the process of mitophagy, inhibiting the senescence of airway epithelial cells, thereby providing a new treatment strategy for COPD.

The invention provides compositions and methods for the treatment of lysosomal storage diseases such as metachromatic leukodystrophy and related disorders. A significant unmet clinical need still exists with the current clinical lentiviral vectors due to the high integration requirement and currently insufficient levels of ARSA expression to correct onset of early symptomatic disease. Methods and compositions for producing a protein such as sulfatase and for treating a disease or disorder such as metachromatic leukodystrophy and related disorders are disclosed.

The invention discloses a method for separating salbutamol in pig urine by using the SLE method, which comprises the following steps: Step 1: sample pretreatment; collect pig urine, first add β-glucuronidase / arylsulfatase to enzymatically After enzymatic hydrolysis, add trichloroacetic acid to precipitate protein in pig urine, add phosphate buffer solution with 4 times the volume of pig urine with a pH range of 6.5 to 7.0, mix with a vortex mixer and centrifuge; take the supernatant after centrifugation; Step 2: After adding the supernatant to the SLE column, let it stand for 2 to 4 minutes; Step 3: Use the binary eluent to elute the SLE column at least twice, receive the eluate, and obtain the eluate containing albuterol . The object of the present invention is to provide a kind of method that adopts SLE method to separate albuterol in pig urine with high separation efficiency, simple operation and low detection limit.

The invention discloses a method for measuring the contents of ractopamine, clenbuterol and salbutamol in swine urine at high precision. The method comprises the following steps of 1, pretreating a sample, namely collecting swine urine, firstly adding beta-glucuronidase hydrochloride / arylsulfatase to carry out enzymolysis, and then, extracting a centrifuged supernatant; 2, adding the supernatant into an SLE column, and then, standing for 2-4 minutes; 3, eluting the SLE column at least twice by using a binary eluting agent, and receiving an eluant to obtain an eluant containing ractopamine, clenbuterol and salbutamol; and 4, detecting the eluant obtained in the step 3 by using a high-performance liquid chromatography. The invention aims at providing the method for measuring the contents of ractopamine, clenbuterol and salbutamol in the swine urine at high precision.

Methods, reagents, and kits for assaying for arylsulfatase A activity for diagnosing conditions associated with arylsulfatase A deficiency, such as multiple sulfatase deficiency (MSD) and metachromatic leukodystrophy (MILD).

The present invention provides, inter alia, methods of treating metachromatic white matter dystrophy disease (MLD) and compositions comprising a recombinant arylsulfatase A (ASA) protein using enzyme replacement therapy.

The invention belongs to the field of medicine and discloses an aryl sulfatase and glucoside aldehydic acid enzyme inhibitor, a preparation method therefor and application of the aryl sulfatase and glucoside aldehydic acid enzyme inhibitor. The preparation method for the aryl sulfatase and glucoside aldehydic acid enzyme inhibitor comprises the following step: adding hypochlorous acid or hypochlorite into a solvent so as to obtain a solution, thereby preparing the aryl sulfatase and glucoside aldehydic acid enzyme inhibitor. Compared with the conventional medical-treatment inhibitor STX64 andinhibitors such as D-Sacchuric acid and 1,4-lactone, the aryl sulfatase and glucoside aldehydic acid enzyme inhibitor has the advantages that the effective inhibition concentration effect is good, inhibition of different degrees can be carried out as required, and the inhibitor has the advantages of simplicity in operation, low cost, rapidness and the like and has a certain application prospect inthe field of medicine.

The present invention pertains to a novel treatment of pathologies caused by an increased synthesis or accumulation of sulfolipids such as sulfatide. The invention provides mutated arylsulfatase A (ARSA or ASA, EC 3.1.6.8) enzymes with increased activity towards sulfatide metabolization. The invention provides nucleic acids encoding the mutant ARSA, the use of the proteins and nucleic acids, as well as pharmaceutical compositions comprising them, in the treatment of lysosomal storage disorders (LSDs) such as metachromatic leukodystrophy (MLD).

The present invention relates to a kit for assaying the activity of the human arylsulphatase or its isozymes, including 4-MUS and 4-MU. The kit can be used for initial screening, therapeutic effect monitoring and recurrence inspecting of human cancers.

The invention belongs to the field of medicine, and discloses an aryl sulfatase inhibitor as well as a preparation method and application thereof. Dodecyl sulfate is added into a solvent, so that a solution can be obtained, namely the aryl sulfatase inhibitor can be prepared. Compared with a conventional medical treatment inhibitor STX64, the inhibitor STX64 has the advantages of being low in effective inhibition concentration, capable of achieving inhibition of different degrees according to needs, easy to operate, low in cost, rapid and the like, and has a bright application prospect in thefields of environment and medicine.

The present invention provides, inter alia, improved methods for purifying arylsulfatase A (ASA) proteins that are recombinantly produced for enzyme replacement therapy. The present invention is based, in part, on the surprising discovery that recombinant ASA proteins can be purified from untreated biological materials (e.g., ASA-containing cell culture media) using processes that include only four chromatographic columns and only one step of post-chromatographic column ultrafiltration / diafiltration.

The invention discloses a method for measuring the content of ractopamine, clenbuterol and albuterol in pig urine with high precision, comprising the following steps: Step 1: sample pretreatment; collecting pig urine, first adding β-glucuronidase hydrochloride / arylsulfatase for enzymatic hydrolysis; take the supernatant after centrifugation; step 2: add the supernatant to the SLE column and let it stand for 2 to 4 minutes; step 3: use binary eluent to clean the SLE column Elute at least twice, receive the eluate, and obtain the eluate containing ractopamine, clenbuterol, and salbutamol. Step 4: Use high performance liquid chromatography to detect the eluate obtained in step 3. The object of the present invention is to provide a method for measuring the contents of ractopamine, clenbuterol and albuterol in pig urine with high precision.

The invention provides a treatment method, a detection method and application of beta-receptor stimulant residues in animal tissues, and belongs to the technical field of food detection. The method for treating the beta-receptor stimulant residues in the animal tissues comprises the following steps: inserting an ultrasonic probe into a first mixed solution containing a sample, an internal standard substance, a buffer solution and beta-glucuronidase / aryl sulfatase, and performing ultrasonic probe-assisted enzymolysis for 1-5 minutes under the condition of output power of 700-1100W. According to the method for treating the beta-receptor stimulant residues in the animal tissues, the extraction efficiency of traditional 16-hour enzymolysis can be achieved within 5 minutes through ultrasonic probe assisted enzymolysis, the treatment time of the beta-receptor stimulant residues in the animal tissues is greatly shortened, and the method has the advantages of being more efficient, more accurate and more convenient.

The invention belongs to the technical field of analysis of toxic organic pollutants, and discloses a method for synchronously and quantitatively determining the concentration of various aromatic compounds and metabolites in urine. The method comprises the following steps: sequentially adding an isotope internal standard substance, a buffer solution and beta-glucuronidase-aryl sulfatase into the urine, carrying out constant-temperature enzymolysis, adding an organic solvent after enzymolysis, extracting, centrifuging, transferring supernatant, concentrating to a constant volume, adding into a polar solution, and carrying out quantification and data processing to obtain the concentration level of various target substances in the urine. By optimizing the scheme, synchronous detection and analysis of various aromatic compounds and metabolites with large property differences are realized, the experimental steps are simplified, the cost is reduced, the sensitivity is improved, and the method can be effectively applied to detection and analysis of large-batch urine samples.

The claimed group of inventions relates to the field of medicine, more precisely to gene therapy for a disease that is incurable as of the date of submission of the application materials - metachromatic leukodystrophy (MLD), which is a severe hereditary neurodegenerative disease that occurs due to deficiency of the lysosomal enzyme arylsulfatase A (ARSA). The essence of the technical solution is a recombinant genetic construct containing CMV / chicken β-actin hybrid enhancer promoter and the codon-optimized ARSA gene sequence represented by SEQ ID NO: 1. Adeno-associated virus of serotype Olig001 for the treatment of metachromatic leukodystrophy, containing a recombinant genetic construct according to claim 1.

The invention discloses a method for effectively purifying aryl sulfatase, which comprises the following steps: (1) adding an alkaline solution into an enzyme solution containing aryl sulfatase and beta-glucuronidase, and adjusting the pH value of the enzyme solution to 11; and (2) adjusting the pH value of the enzyme solution obtained in the step (1) to be neutral by using an acidic solution to obtain a purified aryl sulfatase solution, and almost completely removing residual beta-glucuronidase in the enzyme solution. The method is easy to operate and low in cost, beta-glucuronidase can be specifically removed, meanwhile, more than 85% of aryl sulfatase activity is reserved, non-specific hydrolysis of the estrogen-glucuronate combination can be effectively avoided, the accuracy of the determination result of the estrogen-glucuronate combination in the urine is ensured, and the method is suitable for popularization and application. The method has important application value in medical diagnosis and environmental monitoring research.