23 results about "High mannose" patented technology

The invention discloses an alpha-glucan phosphorylase (alpha GP) mutant. The amino acid sequence of the alpha-glucan phosphorylase mutant is as shown in SEQ ID NO. 3. The invention also discloses an application of the mutant in enzymatic synthesis of hexose, especially mannose. The alpha-glucan phosphorylase mutant provided by the invention has higher catalytic activity, can significantly improve the production efficiency of mannose, and can be used for industrial production of mannose.

The invention relates to the technical field of biology, in particular to a hexanose-2-epimerase mutant and application thereof. According to the hexaldehyde aldose-2-epimerase mutant and the application of the hexaldehyde aldose-2-epimerase mutant in the embodiment of the invention, the hexaldehyde aldose-2-epimerase mutant generates mutation beneficial to improvement of hexaldehyde aldose-2 hydroxyl isomerization catalytic activity at at least one key site in an amino acid sequence as shown in SEQ ID NO: 19; through the mode, the hexaldehyde sacchar-2-epimerase mutant shows excellent hexaldehyde sugar C-2 hydroxyl isomerization catalytic activity, can efficiently convert D-glucose into D-mannose, and is beneficial to improving the synthesis efficiency of the D-mannose.

Provided are compositions comprising glucocerebrosidase having increased phosphorylation of High Mannose and Hybrid N-linked Oligosaccharides. Also provided herein are methods of treating Gaucher disease comprising administering to a subject the compositions of the disclosure.

A high-mannose antibody-specific affinity material, a preparation method therefor and a use thereof. Provided are a lectin variant comprising (a) a histidine tag and (b) a cysteine-containing segment, added to the C-terminus of a reference lectin; a nucleic acid molecule encoding the lectin variant, and a vector and a cell containing the nucleic acid molecule; a composition and a matrix comprising the lectin variant; and a use of the lectin variant for reducing or removing a high-mannose antibody from a sample. The lectin variant has a specific affinity for high-mannose antibodies, and can effectively bind to high-mannose antibodies formed during antibody production, to remove high-mannose byproducts contained in recombinant antibodies, thereby effectively promoting and achieving required antibody purification in downstream stages of production.

Pharmaceutical compositions comprising anti-interleukin 12 (IL-12) interleukin 23 (IL-23) antibodies are described. The pharmaceutical compositions may comprise molecular attributes, such as oxidation of light chain W32, isoaspartic acid at heavy chain position D55; high molecular weight species such as dimers and / or oligomers, high mannose, sialylation, or acidic peak species. Method comprising administering the pharmaceutical compositions are described. Methods of manufacturing the pharmaceutical compositions are described.

The invention belongs to the technical field of biological medicines, and particularly relates to application of alpha-1, 2-high mannose type CD133 as a tumor stem cell sorting marker of intrahepatic cholangiocarcinoma. The lectin Cyanovirus-N (CVN) used in the invention is a 11kDa protein, is derived from Nostoc ellipsoporum, and has a specific recognition function on a terminal alpha-1, 2-Man part on high mannose (Man-8 or Man-9) glycan, the carbohydrate chain structure difference between intrahepatic cholangiocarcinoma tumor stem cells and tumor non-stem cells is studied, and the specific recognition function on the terminal alpha-1, 2-Man part on the non-stem cells of the intrahepatic cholangiocarcinoma tumor stem cells and the non-stem cells of the intrahepatic cholangiocarcinoma tumor non-stem cells is realized. The invention provides a novel marker alpha-1, 2-high mannose type CD133 protein for precise targeting of tumor stem cells of intrahepatic cholangiocarcinoma, and provides application of a novel precise marker for targeting treatment of tumor stem cells by recognizing sugar chain change through lectin.

The present application relates to a method for reducing the level of high mannose type glycoprotein produced by a mammalian cell expression system and its application, the method comprising any one of the following ways or a combination of the two: (a) adding N-acetyl-D-mannosamine to the subculture medium of the previous generation of mammalian cell expression system before inoculation, then inoculating into the production medium after cultivation, and continuing cultivation; (b) adding N-acetyl-D-mannosamine to the culture medium during cultivation after inoculating the mammalian cell expression system from the subculture medium into the production medium. This method can not only effectively reduce the level of high mannose type glycosylation in the produced glycoprotein, but also does not affect the protein yield, and the cell growth and metabolism are also not affected, and other quality results such as protein purity, acid-alkali peak, etc. will not change significantly, so it is significant for the research and preparation of new antibody drugs.

The present invention relates to a cell-free enzyme-catalyzed process for producing glycoproteins of general formula (I) from a lipid-linked oligosaccharide and a peptide. Further, said process includes the construction of the lipid-linked oligosaccharide from a mannose trisaccharide containing core structure. Particularly, the lipid-linked oligosaccharide is a high mannose-, complex-, or hybrid-type N-glycan.

A nucleic acid construct comprising a first polynucleotide encoding MGAT1 gene or a homologous gene having at least 85% sequence identity thereof, a second polynucleotide encoding MGAT2 gene or a homologous gene having at least 85% sequence identity thereof, and a third polynucleotide encoding MAN2A2 gene or a homologous gene having at least 85% sequence identity thereof. A nucleic acid construct combination comprising a first, a second, and a third nucleic acid constructs, which comprise a first polynucleotide encoding MGAT1 gene or a homologous gene having at least 85% sequence identity thereof, a second polynucleotide encoding MGAT2 gene or a homologous gene having at least 85% sequence identity thereof, and a third polynucleotide encoding MAN2A2 gene or a homologous gene having at least 85% sequence identity thereof respectively. A method of regulating the high mannose glycoform content of a recombinant protein during a mammalian cell culture process is also provided, which comprises an expression of the recombinant protein and an overexpression of three proteins that are involved in an N-glycosylation pathway in a mammalian host cell. A highly effective method for reducing the high mannose glycoform content of recombinant proteins in CHO cells.

The present invention relates to methods of upregulating the high mannose glycoform content of a recombinant protein during a mammalian cell culture by manipulating the mannose to total hexose ratio in the cell culture media formulation.

The present invention relates to methods of modulating (e.g., reducing) the mannose content, particularly high-mannose content of recombinant glycoproteins.

The present invention relates to methods of upregulating the high mannose glycoform content of a recombinant protein during a mammalian cell culture by manipulating the mannose to total hexose ratio in the cell culture media formulation.

A method is provided for determining the relative unpaired glycan content of an IgG antibody composition. In an exemplary embodiment, the method comprises (b) treating the IgG antibody composition with two enzymes to form a mixture of Fab fragments and Fc fragments, wherein one enzyme cleaves the antibody heavy chain at the N-terminal site of the hinge region disulfide bond and the other enzyme cleaves the β1,4 bond between core GlcNAc residues to form Fc fragments, each containing a pair of coreglycan structures; (b) separating the Fab fragments from the Fc fragments; and (c) quantifying the abundance of (i) paired non-fucosylated Fc fragments, (ii) unpaired non-fucosylated Fc fragments, (iii) paired high-mannose Fc fragments, and / or (iv) unpaired high-mannose Fc fragments to determine the relative unpaired non-fucosylated (AF) glycan content and the relative unpaired high-mannose (HM) glycan content.

Methods of determining glycan pairing content of a protein composition are provided. In exemplary embodiments, the method comprises (a) treating a sample of the protein composition with (i) a mannosidase and (ii) a β1-4 endoglycosidase, to produce a mixture of components of the protein, wherein at least some of the components comprises one or more digested glycan structures; and (b) separating the components of the mixture based on molecular weight; and (c) quantifying the abundance of glycan pairs, optionally, quantifying (i) paired afucosylated glycans, (ii) unpaired afucosylated glycans, (iii) paired high mannose glycans, and / or (iv) unpaired high mannose glycans, of the mixture. In exemplary aspects, the method comprises quantifying the relative unpaired afucosylated (AF) glycan content and / or relative unpaired high mannose (HM) glycan content of the mixture.

To provide a method for producing anti-IL-36R antibodies, spesolimab.SOLUTION: To provide a method for producing spesolimab using a serum-free cell culture medium in a fed-batch culture in the presence of a reduced copper concentration and an elevated iron concentration. In addition, the present invention provides compositions comprising reduced levels of basic species of Spesolimab and / or reduced levels of Spesolimab species with high mannose structures.SELECTED DRAWING: None

This invention belongs to the field of biotechnology and relates to the application of an endonuclease in the hydrolysis of high-mannose N-glycoproteins. The endonuclease can recognize and hydrolyze high-mannose N-glycoproteins; the endonuclease is Endo-LB, and its amino acid sequence is shown in SEQ ID NO.1; the hydrolysis temperature is 4–60℃, and the hydrolysis pH is 4.0–10.0. Studies have shown that Endo-LB has the activity of recognizing high-mannose N-glycoproteins, and structurally contains a MucBP domain, exhibiting broad temperature and pH adaptability.

The present invention relates to a method for producing the anti-IL36R antibody spesolimab. More specifically, the present invention relates to a method of producing spesolimab with a serum-free cell culture medium in a fed-batch culture in the presence of reduced copper and increased iron concentrations. In addition, the present invention relates to a composition comprising low levels of basic species of spesolimab and / or low levels of spesolimab species with high mannose structures.

The present invention relates to a method for producing the anti-IL36R antibody spesolimab. More specifically, the present invention relates to a method of producing spesolimab with a serum-free cell culture medium in a fed-batch culture in the presence of reduced copper and increased iron concentrations. In addition, the present invention relates to a composition comprising low levels of basic species of spesolimab and / or low levels of spesolimab species with high mannose structures.

Recombinant human alpha glucosidase (rhGAA) compositions derived from CHO cells are disclosed, comprising a more optimal glycan composition consisting of rhGAA with higher amounts of N-glycans bearing mannose-6-phosphate (M6P) or bis-M6P, along with low amounts of non-phosphorylated high mannose glycans, than conventional rhGAA, along with low amounts of terminal galactose on complex oligosaccharides. Compositions comprising the rhGAA are described, as are methods of use.

Recombinant human alpha glucosidase (rhGAA) composition derived from CHO cells that contains a more optimized glycan composition consisting of a higher amount of rhGAA containing N-glycans carrying mannose-6-phosphate (M6P) or bis-M6P than conventional rhGAAs, along with low amount of non-phosphorylated high mannose glycans, and low amount of terminal galactose on complex oligosaccharides. Compositions containing the rhGAA, and methods of use are described

The application discloses a method for improving expression of exogenous glycoprotein by yeast, comprising the following steps: without changing endogenous genes of a yeast host cell and without introducing exogenous sugar chain processing enzymes, an exogenous N-acetylglucosamine transferase I (GnT1) gene from different species is integrated into a genome of a Pichia pastoris host bacterium for expressing glycoprotein, and a Pichia pastoris engineering bacterium for expressing the exogenous GnT1 is constructed, so that the expression amount of the exogenous glycoprotein in the Pichia pastoris engineering bacterium is significantly improved, the proportion of high mannose type N-glycan is reduced, and the proportion of complex type N-glycan is increased. The method is suitable for biosynthesis of polysaccharide glycosylation protein, high isoelectric point protein and complex folding protein, and has good popularization and application prospect in the fields of biological pharmacy, functional protein, nutritional and healthy raw material and industrial biological manufacturing.

The application discloses a kind of based on konjak powder's mannose extraction method, its extraction system includes: konjak powder pretreatment, preparation extraction solvent, separation and filtration, chromatographic separation, mannose purification, crystallization and drying and finished product detection preservation, the preparation extraction solvent includes konjak powder dissolution, enzymatic extraction, the crystallization and drying include mannose crystallization, crystalline separation and crystalline drying.The application can effectively reduce the starch impurities in enzymatic extraction of mannose by the method, thereby effectively improve the extraction quality of mannose, by separation and filtration, chromatographic separation and mannose purification step, can effectively remove the impurities in solution, by chromatographic technique is separated, to purify the mannose therein, by reverse phase high performance liquid chromatography column to the separated mannose is purified, to obtain high-purity mannose, so as to effectively improve the extraction quality of mannose.

The invention discloses a detection kit and a detection method for specifically recognizing a glycosylated MDA5 autoantibody. The method comprises the following steps: adopting a group of recombinant human MDA5 proteins which respectively present specific glycosylated modifications such as a high mannose type, a composite type, a hybrid type and the like as capture antigens; respectively reacting with three antigens to obtain three independent antibody subgroup signals; a matched data processing system integrates three independent signal values into a multi-dimensional glycosylation antibody spectrum (GAP) feature vector through standardized calculation. According to the invention, accurate risk stratification and early warning of high-risk patients with rapid progression type interstitial lung diseases and the like can be realized.