109 results about "Sialidase" patented technology

The present invention relates to eukaryotic host cells which have been modified to produce sialylated glycoproteins by the heterologous expression of a set of glycosyltransferases, including sialyltransferase and/or trans-sialidase, to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. Novel eukaryotic host cells expressing a CMP-sialic acid biosynthetic pathway for the production of sialylated glycoproteins are also provided. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities (such as those involved in sialylation) to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation.

The invention relates to the field of biotechnologies, and provides a construction method for a sialidase gene knockout mouse model and application thereof. An in vitro and vivo targeted mouse genomeNPL is used for genetically recombining a CRISPR knockout plasmid vector, and a diploid double-knockout NPL mouse knockout model is successfully constructed through an oosperm microinjection technology. The NPL has the important function in a pathophysiological process of a body. Vicious transformation of a cancer cell is always along with overexpression of sialic acid. Now, the overexpression ofthe sialic acid has become one of important markers of the malignant transformation and transferring of a tumor. The model is capable of constructing a new research platform for researching a processof cell canceration, beneficial to a key step of researching generation and transferring of the tumor, and providing a foundation for the research and development of a novel anti-tumor drug.

The invention provides a sialidase detection kit. The kit comprises a reagent 1 and a reagent 2, wherein the reagent 1 comprises the components of buffer solution, neuraminidase, lactic dehydrogenase, surfactant, stabilizer and preservative; and the reagent 2 comprises the components of buffer solution, reducing coenzyme, neuraminic acid aldolase, stabilizer and preservative. The kit has accurate detection and low cost and is convenient, applicable, stable and reliable.

The present invention provides compositions and methods for the production of glycoproteins with enhanced sialylation. In particular, the invention provides cell lines comprising disrupted sialidase expression and methods of using the cell lines to produce glycoproteins with enhanced sialylation.

This invention discloses a sialic acid enzyme test reagent for testing sialic acid enzyme activity in vagina secreta outside the body, containing substrate on the carrier named N-acetyl neuraminate and its salt which can be its derivant or thymolphthaleic N-acetyl neuraminate and its salt, 5-Br-4-Cl-3-indolyl-alpha-D-N-acytylneuraminate and its salt. We can diagnose the bacteriogenic vagina deseases quickly and simple by testing sialic acid enzyme to vagina searate with this sialic acid ester reagent which can be used as an independent diagnosis target with good stability, extremely excellent accuracy and simple operation (one step test).

This disclosure provides reagent to treat tumor and cancer, and methods of using the same for treating tumor and cancer. One type of the reagent is pro-antibody, which is antibody that can be activated in tumor. Another type of the reagent is a conjugate of sialidase with affinity ligand that can bind to immuno cell surface or a conjugate of sialidase with affinity ligand that can bind to another antibody, therefore provide a sialidase based cancer immuno therapy strategy.

The invention provides a lung cancer monitoring kit and a use method thereof. The kit includes: a reagent A, which is prepared by adding 5% of SDS to a 10 mM ammonium bicarbonate solution; a reagent B, which is prepared by adding not less than 2.2 unit/[mu]L of exoglycosidase to NP40 being 3.3% in concentration; a reagent C, which is a mixture liquid composed of, in equal volume, a solution of 20 mM of APTS dissolved in 1.2 M of citric acid and a solution of 1M of an organic reducing agent dissolved in DMSO; a reagent D, which includes 0.25 [mu]L of NH4Ac with concentration being 100 mM and pH value being 5, 0.2 [mu]L of sialidase, and 2.55 [mu]L of H2O2. The kit can evaluate lung cancer patients with detection on a fingerprint spectrum of an oligosaccharide chain in blood glycoprotein as a diagnosis index, thereby diagnosing the tumor stage of the lung cancer. The kit allows numerous lung cancer patients to take regular and non-invasive detection, and helps a medicinal worker to detect the lung cancer and monitor progression of the lung cancer timely.

A preparation method of sialidase comprises the following steps: preparation of a sialidase gene fragment; cloning of the sialidase gene fragment into pMD-18T to obtain a connection product, conversion of the connection product into E.coliJM109, and selection of a positive clone for sequencing to obtain a gene sequence as shown in SEQIDNO:1; connection of the positive clone and a prokaryotic expression vector to obtain a recombinant expression vector; conversion of the recombinant expression vector to obtain recombinant engineering bacteria; fermentation, induced expression and purification of the recombinant engineering bacteria to obtain the sialidase. The preparation method of the sialidase comprises bacterial genome DNA extraction, PCR amplification, construction of the recombinant expression vector, conversion of the recombinant expression vector into host cells for construction of recombinant expression cells, culture and induced expression of the recombinant expression cells, isolation and purification of protease from the cultured host cells, and analysis of protease enzymatic properties. The sialidase prepared by the preparation method has high temperature resistance and Ph resistance, and provides more basic materials and scientific bases for industrial and social application.

The invention relates to a diagnostic kit for early liver cancer. The diagnostic kit comprises the following components of G1: 5% of SDS (sodium dodecyl sulfate); G2: 2.2 microgram/muL glycanase; G3A: 100mM trisulphonate fluorescent marker; G3B: 1M organic matter reducing agent; G4: sialidase; GW: deionized water. The invention also discloses a use method of the diagnostic kit. The diagnostic kit has the advantages that the problem of high missed diagnosing rate of alpha fetoprotein on the liver cancer is solved; the liver cancer can be quickly, simply, conveniently and accurately detected, and the therapy effect of the liver cancer can be tracked.

The invention provides a method for measuring sialic acid content of protein. The method comprises the steps of 1, using sialidase to conduct digestive treatment on a protein sample, so that digestive juice containing sialic acid is obtained; 2, conducting sedimentation treatment on the digestive juice, and separating supernatant containing sialic acid; 3, using an ultra-high performance liquid chromatography-mass spectrometry method to analyze the supernatant, so that a mass spectrogram is obtained; 4, based on the mass spectrogram, determining the sialic acid content of protein. In the method for measuring the sialic acid content of protein, on the condition that there is no need to conduct derivatization on sialic acid, the total sialic acid content in the protein sample is subjected to rapid quantitative detection, the method is easy to operate, the sensitivity is high, the accuracy is high, the elapsed time is short, and the method can be used for intermediate quality control over biomacromolecule production or quality inspection of final products.

The present invention describes compounds of Formula I or a pharmaceutically acceptable salts or derivatives thereof. Compositions comprising compounds of Formula I are also described. The present invention further relates to a method of producing non-2-enonate compounds.

A vaccine against the Chagas disease, capable of stimulating the immune response against the trans-sialidase virulence factor of the Trypanosoma Cruzi parasite, which is a multicomponent vaccine comprising: (a) an immunogenic portion formed by one or more recombinant or synthetic polypeptides or fractions of thereof and (b) one or more polynucleotides including the regions that codifying one or more immunogenic polypeptides, or a monocomponent comprising at least one component selected among an immunogenic portion formed by one or more recombinant or synthetic polypeptides or fractions of them and a group of polynucleotides including the regions codifying one or more immunogenic polypeptides derived from Trypanosoma Cruzi and pharmaceutical compositions containing said multicomponent and monocomponent vaccines, the procedures for obtaining the immunogen portion of said vaccines and the nucleic acid used in the procedure.