65 results about "Protein glycosylation" patented technology

The invention provides methods and compositions for the rapid and sensitive detection of post-translationally modified proteins, and particularly of those with post-translational glycosylations. The methods can be used to detect O-GlcNAc posttranslational modifications on proteins on which such modifications were undetectable using other techniques. In one embodiment, the method exploits the ability of an engineered mutant of β-1,4-galactosyltransferase to selectively transfer an unnatural ketone functionality onto O-GlcNAc glycosylated proteins. Once transferred, the ketone moiety serves as a versatile handle for the attachment of biotin, thereby enabling detection of the modified protein. The approach permits the rapid visualization of proteins that are at the limits of detection using traditional methods. Further, the preferred embodiments can be used for detection of certain disease states, such as cancer, Alzheimer's disease, neurodegeneration, cardiovascular disease, and diabetes.

The invention relates to methods for identifying compounds which affect cellular stress. In particular, the method relates to identifying compounds which inhibit protein advanced glycation end product formation, where the compounds are carbonyl scavengers which inhibit the formation. The assay involves combing the substance of interest with histone H1 and ADP-ribose, and then measuring fluorescence and protein cross linking. Various inhibitors ofprotein AGE glycation have been identified, using this assay.

The invention relates to a method for analyzing protein O-glycosylation sites. The method comprises the following steps: (1), sequentially carrying out endonuclease digestion, endoglycosidase and exoglycosidase digestion on a proteome sample or a single protein sample to prepare a peptide and non-glycopeptide protein sample or single protein sample with a GalNAc label; (2) loading the sample on a durian agglutinin chromatographic column and separating to prepare a proteomic sample; (3) separating the single protein sample or the proteomic sample through with a C18 reversed-phase chromatographic column, and then detecting by a high-definition mass spectrometer in a cation mode to obtain a high-definition mass spectrogram; (4) processing the obtained mass spectrometric data and obtaining protein glycosylation site information though the searched result. The site information can be more rapidly, more accurately and more comprehensively obtained through establishing the method for detecting the O-glycosylation sites of glycoproteins in the single protein sample and the proteomic sample.

The present invention relates to methods and compositions for the identification of cancer markers. In particular, the present invention provides methods and compositions for the identification of glycosylated proteins and protein glycosylation patterns. The present invention further provides cancer markers identified using the described methods.

The invention discloses a kit for detecting protein glycosylation subtype and a detection method thereof. The detection kit comprises agglutinin, confining liquid, a biotin labeled antibody, horse radish peroxidase labeled streptavidin, a 3',3',5',5'-tetramethyl benzidine substrate developing solution, a stop solution and a cleaning buffer solution. The detection method comprises the following steps: fixing the agglutinin onto a polystyrene solid phase carrier to be used for recognizing and binding glycoprotein in the sample based on specific recognition of agglutinin and an oligosaccharide chain, recognizing the glycoprotein needing to be detected by virtue of specific binding of the protein and the antibody thereof, and realizing semi-quantitative analysis of the glycoprotein. According to the detection kit and the detection method disclosed by the invention, semi-quantitative detection of single protein glycosylation subtype can be realized, and the kit is easy and convenient to operate and low in cost.

The invention belongs to the field of biotechnologies, and relates to a method for applying recombinant expression separated and purified human-derived protein Fn3 (fibronectin type III domain) mutants as N-glycosylation efficiency detection model proteins. The method includes steps of constructing Fn3-Gly-loop recombinant protein gene expression vectors of the Fn3 mutants; jointly transforming the constructed expression vectors into Escherichia coli engineering strains CLM37 by means of electric shock processes; screening the expression vectors by the aid of antibiotics to obtain positive clones. Recombinant proteins contain Fn3-Gly-loop proteins which are about to be modified by recombinant glycosyl, and the Fn3-Gly-loop fusion protein glycosylation efficiency can be detected by the aid of Western Blot processes. The method has the advantages that the skeleton proteins Fn3 in Escherichia coli are used as receptor proteins, accordingly, the model receptor proteins suitable for N-glycosylation modification efficiency research can be constructed, and the recombinant protein glycosylation efficiency can be easily, quickly and efficiently detected in the Escherichia coli.

The invention improves glycoprotein production and protein glycosylation engineering in eukaryotes, specifically the production of human-like complex or hybrid glycosylated proteins in lower eukaryotes such as yeasts. The invention provides glycosylation modified eukaryotic host cells capable of producing glycosylation optimized proteins useful as immunoglobulins and other therapeutic proteins, and provides cells capable of producing glycoproteins having glycan structures similar to glycoproteins produced in human cell. The invention further provides proteins with human-like glycan structures and novel compositions thereof producible by these cells.

The disclosure relates to an oligosaccharyltransferase polypeptide and the production of glycosylated recombinant protein in a microbial host cell; and including vaccines comprising glycosylated recombinant antigens.

The invention discloses a method for preparing beta-carotene uniform emulsion from whey separation protein glycosylation reaction products and the uniform emulsion. The method comprises the followingsteps: 1) preparing the whey separation protein glycosylation reaction products; 2) preparing a glycosylated protein-polyphenol compound; and 3) preparing the beta-carotene uniform emulsion. The goodemulsifying property and interface property of the glycosylated protein-polyphenol nano-compound effectively inhibit the coalescence phenomenon among oil drops, the emulsion drops show good uniformity, and the emulsion drop uniformity is directly related to the storage stability. Meanwhile, the glycosylation reaction enables protein to form a denser and thicker interface layer outside the oil drops loaded with active substances, so that the interaction between the active substances and an oxidant (metal ions, free radicals and dissolved oxygen) is inhibited.

The invention discloses a vaccine for preventing and treating diseases caused by Acinetobacter baumannii and a preparation method thereof. The vaccine for preventing and treating the diseases caused by the Acinetobacter baumannii provided by the invention has an active component which is an Acinetobacter baumannii glycoprotein obtained through glycosylation of a protein composed of bases from site20 to site 156 of SEQ ID No. 1 or bases from site 1 to site 156 of SEQ ID No. 1. According to the invention, Acinetobacter baumannii glycoprotein modified rCTB4573 prepared by a genetic engineering method is used for preparation of an Acinetobacter baumannii glycoprotein protein conjugate vaccine; uniformity of the vaccine can be improved; production efficiency of the vaccine is improved; the cost is reduced; extensive application prospect is achieved; and the vaccine can be used for preventing the diseases caused by pathogenic Acinetobacter baumannii.

The present invention relates to methods for glycosylating a protein in which the protein is modified to include an alkyne and/or an azide group. The invention further relates to a protein glycosylated by these methods.

The present disclosure provides methods for improving batch-to-batch consistency of proteins produced by eukaryotic cells, as well as proteins produced thereby.

The present invention relates to pancreatic cancer markers. In particular, the present invention provides methods and compositions for the identification of protein glycosylation patterns associated with pancreatic cancer.

Methods for producing isotopically-labelled peptides are provided. Aspects of the method include: contacting a sample including a metabolically tagged protein with a cleavable probe to produce a probe-protein conjugate; separating the probe-protein conjugate from the sample; digesting the probe-protein conjugate to produce a probe-peptide conjugate; and cleaving a cleavable linker to release an isotopically labelled peptide. The method may further include: identifying a predetermined isotopic pattern in a mass spectrum; determining an amino acid sequence of the isotopically labelled peptide; and identifying the site of protein glycosylation based on the determined amino acid sequence. Also provided are cleavable probes for practicing the subject methods, described by the Formula: A-L-(M-Z) where A is an affinity tag, L is a cleavable linker, M is an isotopic label and Z is a chemoselective tag capable of cross-linking a metabolically tagged protein. Compositions and kits for practicing the subject methods are also provided.

The invention relates to an artemisia selengensis straw extractive, a preparation method and a function of inhibiting protein glycosylation. The preparation method of the extractive comprises the following steps of: by taking the artemisia selengensis straw as the raw material, carrying out reflux extraction by using 75% of ethanol with the solid-liquid ratio of 1:15 at 90 DEG C, filtering, carrying out decompression concentration on the filtrate, extracting and decoloring with petroleum ether, further purifying macroporous resin AB-8 column chromatography, eluting the chromatography column by 10% of water, 30-50% of water and 95% of ethanol, collecting 30-50% of eluted component of ethanol for decompression concentration and carrying out vacuum drying to obtain solid powder. The total flavone content of the artemisia selengensis extractive is not less than 80% and the main components comprise rutin and luteolin; the artemisia selengensis straw extractive has the activities of capturing active dicarbonyl compound (methylglyoxal MGO and glyoxal GO) and inhibiting the protein glycosylation (AGEs).

The application provides an immune composition and a genetically engineered subunit protein vaccine. The vaccine includes newcastle disease virus HN protein encoded by the nucleic acid molecule shownin SEQIDNO:1 or nucleic acid molecule with the nucleic acid sequence has more than 95% similarity with that of the nucleic acid molecule shown in SEQIDNO:1, and newcastle disease virus F protein encoded by the nucleic acid molecule shown in SEQIDNO:3 or nucleic acid molecule with the nucleic acid sequence has more than 95% similarity with that of the nucleic acid molecule shown in SEQIDNO:3. The vaccine adopts CHO cell eukaryotic expression, protein glycosylation is thorough, the antigen protein immunogenicity is good, the expression quantity is very high and up to (2-3) g/L, recombinant cellscan be subjected to large-scale suspension culture, the complexity of vaccine preparation is reduced greatly and the production cost is reduced.