33 results about "Glycoproteomics" patented technology

The present disclosure relates to tailored glycoproteomic methods, and more particularly to methods for the sequencing, mapping and identification of cellular glycoproteins using saccharide-selective bioorthogonal probes. A method is disclosed for saccharide-selective glycoprotein identification (ID) and glycan mapping (GIDmap) that generates glycoproteins tailored with bioorthogonally tagged alkynyl saccharides that can be selectively isolated, allowing for glycoprotein ID and glycan mapping via mass spectromic proteomics, including liquid chromatography-tandmen mass spectroscopy (LC-MS2). LC-MS2 may be used to identify cellular glycans, and more specifically cancer-related glycoproteins.

A cysteine hydrazide nicotinamide (Cyhn) reagent designed for the enrichment of bacterial glycoproteins is provided. Methods for purification of free oligosaccharides and their analysis are also provided.

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 a polymer chain-modified silica gel matrix hydrophilic interaction chromatographic stationary phase and a preparation method and application thereof. Using silica gel particles as the substrate, graft hydroxyl-containing ethylenic monomers and methylenebisacrylamide on the surface of silica gel through the click copolymerization reaction of thiol-ene to form a surface with three-dimensional dendritic polymer chains rich in hydroxyl and amide affinity. Hydrophilic interaction chromatography stationary phase with aqueous functional groups. The introduction of hydrophilic functional groups such as hydroxyl and amides according to the method of the present invention not only overcomes the shortcomings of traditional post-modification methods, such as tedious steps and low reaction efficiency, but also the three-dimensional polymer chain structure on the surface can form a rich hydrophilic layer , has a better advantage as a hydrophilic chromatography stationary phase. The liquid chromatography stationary phase provided by the invention has a novel structure and excellent hydrophilicity, and can be widely used in the separation of various samples and the selective enrichment and separation of glycopeptides, and has great potential in the fields of separation analysis and glycoproteomics. Strong practical value.

The invention relates to preparation of chitosan hydrophilic macroporous monolithic material by freeze-drying method and application thereof. The specific preparation process uses chitosan as a monomer, polyethylene glycol diglycidyl ether as a crosslinking agent, and acetic acid solution as a solvent to prepare the chitosan hydrophilic monolithic material by freeze-drying. The overall material of the present invention is low in cost, green and environmentally friendly, has good specific adsorption effect on glycopeptides in complex samples, and has strong application potential in glycoproteomics research.

A diagnostic method for determining the absence or presence of a disease is provided. The method includes assaying the amount and / or types of glycopeptides in a sample from a subject, and comparing these to the amount and types of reference glycopeptides. The method may include the use of a stable isotope label, affinity selection, immunoaffinity chromatography, and glycoproteomics techniques, to identify and quantify changes in glycosylated peptides or glycosylated proteins associated with cancers such as malignant lymphoma or breast cancer, to monitor patient's response to therapy, and to monitor disease recurrence.

The invention relates to a bonding polysaccharide type hydrophilic chromatographic stationary phase as well as a preparation method and an application thereof. A structure of the bonding polysaccharide type hydrophilic chromatographic stationary phase is shown in the specification, wherein SiO2 is silica gel, saccharide is polysaccharide with molecular weight ranging from hundreds to tens of thousands. The invention provides the preparation method of the stationary phase material, wherein the preparation method comprises the following step: modifying the surface of the silica gel and bonding with the polysaccharide in an aqueous liquor system by taking carbonyl dimidazole (CDI) as a connecting arm to prepare the bonding polysaccharide type chromatographic stationary phase. Compared with the conventional bonding saccharide type stationary phase preparation method, the stationary phase preparation method disclosed by the invention is more efficient, convenient and environment-friendly. And meanwhile, the bonding polysaccharide type hydrophilic chromatographic stationary phase has good separating selectivity, repeatability and stability, has great application potentiality in separating polar compounds of other types, and is widely applied in saccharide separating and saccharide-modifying proteomics.

The invention discloses a lectin simulant preparation method based on a molecular imprinting technique. The preparation method includes the steps: firstly, acquiring a carbohydrate chain on complete glycoprotein as a template molecule by enzyme digestion reaction; secondly, fixing the template molecule on the surface of a boric acid functional substrate material by the aid of a boric affinity function; thirdly, performing condensation reaction by the aid of silanization reagents to form an imprinting layer; finally, removing a template under the acid conditions to form an imprinting cavity to obtain a lectin simulant based on molecular imprinting. The lectin simulant is simple to prepare, low in cost, stable in nature, excellent in specificity and affinity and high in anti-jamming capacity, a recognized target object is more easily released, integrated glycoprotein can be recognized, characteristic fragments such as glycopeptides and glycan of the glycoprotein can be recognized, and the lectin simulant still can keep single-minded recognized capacity in used for complex biological samples and has an excellent application prospect in aspects such as glycoproteomics, metabonomics, glycobiology, disease diagnosis and analeptic inspection.

The invention relates to a preparation method and an application of a hydrophilic chromatographic stationary phase of a cationic polysaccharide coating type, wherein silica gel is provided with a negative charge; saccharide is of a quaternary ammonium polysaccharide provided with a positive charge. According to the preparation method, by utilizing the ionic interaction, the silica gel with a sulfonated surface and the cationic polysaccharide with the positive charge are combined in an aqueous solution system so as to prepare the agglomeration type chromatographic stationary phase. Compared with the traditional bonding sugar type stationary phase, the preparation method of the agglomeration type stationary phase is more efficient, convenient and environment-friendly. In addition, the agglomeration type stationary phase has good separation selectivity, repeatability and stability, thereby having the great application potentiality for the separation of polar compounds of other types. Therefore, the agglomeration type stationary phase can be widely applied to glycobiology and glycoproteomics.