155 results about "Phosphorylated Peptide" patented technology

The invention discloses a synthesis method of a metal organic framework composite material which has a sandwich structure, comprises magnetic microsphere ferroferric oxide surface coating polydopamine and takes zirconium ion as central metal ion and application of the material. The synthesis method comprises the following steps: firstly synthesizing ferroferric oxide magnetic microspheres by using a hydrothermal synthesis method; dispersing the magnetic microspheres in an aqueous solution of polydopamine hydrochloride to coat the surfaces of the magnetic microspheres with polydopamine layers; dispersing the magnetic microspheres coated with the polydopamine in an N,N-dimethylformamide mixed solution of zirconium chloride and terephthalic acid to prepare the magnetic metal organic framework composite material with a sandwich structure. The synthesis method is simple and quick; the magnetic metal organic framework composite material obtained by synthesizing is high in specific surface area, good in biocompatibility and applicable to selective enrichment of phosphorylated peptide and MALDI-TOFMS detection in biological samples.

The invention provides a synthetic method and application of a metal-organic framework (MOF) composite nanomaterial. The method comprises the following steps: dispersing ferriferrous oxide magnetic spheres which are synthesized through a traditional hydrothermal technology in a weakly alkaline solution of dopamine hydrochloride to carry out self-polymerization of dopamine on the surfaces of the magnetic spheres; and sequentially dispersing polydopamine coated magnetic spheres in a dimethylformamide solution of zirconium chloride and a dimethylformamide solution of 2-amino-terephthalic acid to obtain the MOF composite nanomaterial with the magnetic sphere surfaces coated with polydopamine and modified with an amino group and with zirconium as a center metal ion. The material has the advantages of large specific surface area, good hydrophilicity and suitable pore structure, can be applied to further researches of the proteomics, and can specifically enrich Which can specifically enrich phosphorylated peptide segments and glycopeptides; the synthetic method is simple and quick; and the synthesized material has good hydrophilicity and biocompatibility, and can be used for selectively enriching endogenous phosphorylation peptide segments and glycopeptide in complex biological samples.

The present invention describes novel tumor-specific phosphorylated peptides, nucleic acids encoding those peptides, and antibodies generated against said peptides. The genes, peptides, and antibodies described herein may be used as diagnostic indicators of the presence of cancer and / or used in therapeutics to treat cancer.

The invention provides a method for specifically separating and enriching a phosphorylated peptide and a glycosylated peptide. The method concretely comprises the following steps: preparing a dispersion from a hydrophilic magnetic meso-porous titanium dioxide material, adding the dispersion and a target phosphorylated peptide and glycosylated peptide solution into a sample introduction buffer solution, performing incubation at 37 DEG C for 30-60 min, washing a material with the sample introduction buffer solution, using ammonia water with the volume ratio of 5-20% as an elution buffer solution, performing dot targeting on the obtained eluate, and performing mass spectrometry. The method realizes the simultaneous enrichment of the low-abundance phosphorylated peptide and glycosylated peptide by controlling enriching and eluting conditions, can realize simultaneous large-scale identification of a phosphorylated protein and a glycosylated protein by MALDI-TOF MS or nano-LC-MS / MS, and hasa broad application prospect in post-translational modification proteomics.

There are provided a novel compound which captures a multisite phosphorylated peptide or protein specifically to a phosphorylation site and a method for detecting a multisite phosphorylated peptide or protein using the same. In particular, there are provided a compound which specifically detects an excessively phosphorylated tau protein observed in the brain affected by Alzheimer's disease and a method for diagnosing Alzheimer's disease in vitro or in vivo using the compound. By bringing a metal complex compound having two dipicolylamine (Dpa) moieties and a spacer including a chromogenic or luminescent functional group or atom group into contact with a multisite phosphorylated peptide or protein, the compound recognizes the distance between phosphate groups and specifically binds to the peptide or the protein, and a multisite phosphorylated peptide or protein or a kinase activity is optically detected by measuring the change, or a multisite phosphorylated peptide or protein or kinase activity is imaged by an optical imaging method applying the change in the luminescence.

The invention relates to polydopamine modified magnetic balls, a synthetic method of nanomaterial by fixing Ti<4+> on the surfaces and an application thereof. The synthetic method comprises the following steps: dispersing magnetic balls in a dopamine-contained water solution, quickly adding a trismetyl aminomethane buffer solution in the dopamine-contained water solution in which the magnetic balls are uniformly dispersed under the mechanical stirring condition, mechanically stirring for 10 to 16 hours at a room temperature, and washing and carrying out the magnetic separation to obtain nanomaterial which is good in dispersivity in the water and provided with the polydopamine modified magnetic balls on the surfaces, and dispersing the material in a 100 mmol / L Ti(SO4)2 solution, mechanically stirring for 2 to 3 hours at the room temperature, and washing and carrying out the magnetic separation to obtain polydopamine modified magnetic balls, and nanomaterial by fixing Ti<4+> on the surfaces of the magnetic balls.. The method for fixing Ti<4+> is very simple, a layer of polydopamine is modified on the surface, so that the material has excellent environmental stability and biocompatibility, and is good in dispersivity in the water, and the material can fix a large amount of Ti<4+>, and takes the magnetic balls as a core to cause the material to be simple to operate and carry out magnetic separation, the synthetic method is simple, and low in cost, and can enrich phosphorylated peptide by a high-flux MALDI-TOFMS.

The present invention is a method, and a kit for carrying out the method, of separating phosphorylated peptides from a mixture of phosphorylated and unphosphorylated peptides. The method involves reacting a collection of peptides, in which some of the peptides have one or more phosphate groups, with a first resin. The peptides are then reacted with a capture ligand which reacts with the phosphate group of the phosphorylated peptides to form a bond. The capture ligand is used to separate the phosphorylated peptides from the unphosphorylated peptides.

The invention relates to a preparation method of a hydrophilic metal ion immobilization affinity magnetic bead, which comprises the following steps: the surface of an amino magnetic bead is subjected to amino activation through glutaral; the activated magnetic bead is connected with an adenosine disodium triphosphate molecule through a surface modification reaction; the surface of the activated magnetic bead is bonded with a hydrophilic functional molecule containing a plurality of phosphate and hydroxyl radicals, so as to be beneficial for subsequent metal ion immobilization; at last, the immobilization of a metal ion is realized in a solution containing the metal ion; and the preparation of the hydrophilic metal ion immobilization affinity magnetic bead is accomplished. The hydrophilic metal ion immobilization affinity magnetic bead has the advantages that the preparation method is simple; compared with other metal ion immobilization affinity chromatographic columns or magnetic beads, the hydrophilic metal ion immobilization affinity magnetic bead prepared by the method is high in hydrophily, good in selectivity, good in preparation repeatability and stable in performance, and has a good specific concentration ability to phosphorylated peptides; the operation is simple; the complicated centrifugation operation can be avoided; and the hydrophilic metal ion immobilization affinity magnetic bead has wide application prospects in proteomics, especially in phosphorylated proteomics.

A set of phosphorylated peptides are presented by HLA A*0101, A*0201, A*0301, B*4402, B*2705, B*1402, and B*0702 on the surface of melanoma cells. They have the potential to (a) stimulate an immune response to the cancer, (b) to function as immunotherapeutics in adoptive T-cell therapy or as a vaccine, (c) to facilitate antibody recognition of the tumor boundaries in surgical pathology samples, and (d) act as biomarkers for early detection of the disease. Phosphorylated peptides are also presented for other cancers.

We characterized a total of 175 HLA-DR-associated phosphopeptides using sequential affinity isolation, biochemical enrichment, mass spectrometric sequencing and comparative analysis. Many were derived from source proteins which may have roles in cancer development, growth and metastasis. Most were expressed exclusively by either melanomas or transformed B cells, suggesting the potential to define cell type-specific phosphatome “fingerprints”. We generated HLA-DRβ1*0101-restricted CD4+ T cells specific for a phospho-MART-1 peptide identified in two melanoma cell lines. These T cells showed specificity for phosphopeptide-pulsed antigen presenting cells as well as for intact melanoma cells. MHC II-restricted phosphopeptides recognizable by human CD4+ T cells are potential targets for cancer immunotherapy.

Provided are compositions containing amorphous calcium carbonate (ACC), and at least one phosphorylated peptide which stabilizes the amorphous form of said calcium carbonate. Particularly, the peptide can be selected from crustacean proteins, also provided by the invention, namely GAP65, GAP22, GAP21, and GAP12 (also indicated herein as GAP10). The compositions are useful in pharmaceutical and nutraceutical formulations.

The present invention provides methods and materials for isolating, purifying, and / or enriching the concentration of compounds having one or more phosphate groups and / or derivatives thereof, including but not limited to phosphorylated peptides and / or phosphorylated proteins. In some aspects, the present invention provides nanostructured enrichment materials, such as metal oxide mesoporous materials, that selectively and reversibly bind with phosphorylated compounds with high specificity and are capable of controlled release of phosphorylated compounds bound to their active surfaces. Mesoporous materials of the present invention also provide enrichment materials having large active surface areas that provide for higher loading capacities for phosphorylated peptides and proteins relative to conventional affinity based methods. Nanostructured metal oxide mesoporous enrichment materials of the present invention are also compatible with implementation via a variety of separation platforms including flow through separation systems, elution based separation systems, column chromatography and affinity chromatography.

The invention relates to the enrichment and purification of phosphopeptide in phosphoproteome, in particular to the application of ZrO2 in the enrichment and purification of phosphopeptide. ZrO2 nano material shows high specificity, selectivity, binding capacity and sensitivity at a phosphopeptide section; meanwhile, the affinity of the ZrO2 nano material to the specificity of phosphopeptide ensures that the material can be used in phosphoproteome during protein post-translational modification. Due to the specificity combination of ZrO2 with the phosphopeptide section in complex proteolysis sample, large-scale identification of phosphorylated protein and determination of phosphorylation sites can be realized through combining with MALDI-TOF MS and nano-LC MS / MS.

The invention provides a peptide identification method by using a mesoporous silica composite combined with mass spectrum. A composite with perfluoroalkyl group modified mesoporous silica covering on the surface of Fe3O4 magnetic microsphere is used in matrix-assisted laser desorption ionization-time of flight mass spectrometric analysis of perfluoroalkyl group derived peptide. The material shows extremely strong affinity with the fluorous phase derived phosphorylated peptides, while maintaining good dispersion in aqueous solution. The Fe3O4 magnetic core in the material simplifies the process of enrichment and separation. A solid phase microextraction method based on the composite with perfluoroalkyl group modified mesoporous silica covering on the surface of Fe3O4 magnetic microsphere achieves selective enrichment of non-phosphorylated peptides, and can be successfully applied to the enrichment of endogenous phosphopeptides in human serum.

Disclosed is a fluorescent sensor for phosphate ion and phosphorylated peptide, which comprises a phosphate anion-selective fluorescent compound expressed by the following general formula (1). In the formula (1), FL represents a fluorescent functional group or atomic group having an aromatic ring or heterocyclic ring (for example, dimethylanthlyl group), and X represents a functional group or atomic group which will be liberated in an aqueous solution to form an anion (for example, NO3):

The invention discloses a preparation method for a fluorescent molecular imprinted probe. Cadmium telluride quantum dots are taken as a fluorescent element, and serve as a carrier after being subjected to silicon coating treatment, template molecules are replaced by phenyl phosphonic acid, and surface imprinting and antigen determinant methods are used for synthesizing the molecular imprinting probe. The method comprises the following steps of preparing a cadmium telluride quantum dot solution; preparing quantum dot silicon-coated microspheres; preparing a composite material taking the quantum dot silicon-coated microspheres as a core and a molecular imprinted layer as a shell; and preparing the fluorescent molecular imprinting probe. The method has the advantages that high sensitivity of the quantum dots and high selectivity of molecular imprinting are combined by the fluorescent molecular imprinted polymer, the surface of the carrier is modified by the molecular imprinted layer, and recognition sites are close to the surface, so that target molecules can be easily and selectively recognized; the fluorescent probe synthesized by the antigen determinant method can be used for recognizing the target molecules tyrosine phosphorylated peptide, and fluorescence enhancement effects are achieved.

A method for the separation and assay of phosphorylated peptides or proteins from complex mixtures of proteins and peptides is described. The method uses a diazo moiety linked by an organic group to a substrate. Kits for performing the assay are also described. The method and kits are particularly useful for detecting changes in phosphorylates produced in living cells.

The invention relates to preparation of an immobilized metal ion affinity chromatographic material, and particularly relates to a method for preparing an immobilized metal affinity ion chromatographic monolithic column. In the method, a monolithic column material is prepared by using an organic-inorganic hybrid silica gel monolithic material of which the surface carries aldehyde groups as a vector; a phosphate buffer solution containing aminomethylphosphonic acid and sodium cyanotrihydroborate NaCNBH3 flows through the monolithic column material continuously to prepare a monolithic material vector of which the surface carries aldehyde chelation groups; and an aqueous solution of transition metal ions flows through the material vector to prepare the immobilized metal ion affinity chromatographic monolithic column. High-selectivity enrichment of phosphorylated peptide in a hybrid peptide section can be realized by utilizing the immobilized metal ion affinity chromatographic monolithic column. The invention has the advantages that the monolithic column has good selectivity to the phosphorylated peptide, can be recycled, and has the characteristics of large enrichment capability and high recovery rate; and the preparation method has the characteristics of simplicity, convenience, good permeability and small back pressure, and the prepared immobilized metal ion affinity chromatographic monolithic column has good mechanical strength and strong acid and alkali resistance.

According to the present invention, phosphorylated peptides and / or phosphorylated proteins are specifically separated. A sample containing a phosphorylated peptide and / or a phosphorylated protein is supplied to a separation unit filled with a metal oxide in the presence of an aliphatic hydroxycarboxylic acid. Upon separation of a phosphorylated peptide and / or a phosphorylated peptide with the use of a separation unit filled with a metal oxide, adsorption of carboxylic acid to an acidic peptide can be prevented in the presence of aliphatic hydroxycarboxylic acid. In addition, aliphatic hydroxycarboxylic acid does not inhibit adsorption of a phosphorylated peptide and a phosphoric acid group in the phosphorylated peptide to a metal oxide.

The present invention relates to an N-phosphorylated peptide enrichment method. According to the method, O-phosphorylated peptide interference in a peptide sample is removed through beta-elimination, and the N-phosphorylated modified peptide in the sample is enriched by using a magnetic mesoporous titania composite nanometer material; the O-phosphorylated modification on serine and threonine is removed through a beta-elimination method under an alkaline condition so as to form stable dehydroalanine and beta-methyl dehydroalanine; a magnetic Fe3O4 nanometer material is synthesized through a solvent method, titanium dioxide is wrapped on the surface through a sol-gel method, and hole amplification is performed through a hydrothermal method to obtain the magnetic mesoporous titania composite nanometer material; and the magnetic mesoporous titania composite nanometer material can specifically enrich the N-phosphorylated peptide under a weak acid buffer system. According to the present invention, the beta-elimination and specific enrichment combined method is used for enriching the N-phosphorylated peptide in the sample.

The invention discloses a preparation method of a flexible metallic oxide nanofiber phosphorylated peptide enrichment material. The preparation method of the flexible metallic oxide nanofiber phosphorylated peptide enrichment material includes the steps that 1, metal salt is added into corresponding solvent, stirring is conducted to generate metal ions, then, a chelating agent is added, stirring is conducted again, and a precursor solution is obtained, wherein the ratio of the metal salt to the solvent is 1 g:(10-80) ml, and the molar ratio of the metal salt to the chelating agent is 1:(0.01-0.4); 2, the obtained precursor solution is subjected to electrostatic spinning to obtain a precursor nanofiber membrane; 3, the precursor nanofiber membrane is calcined in air, the calcining temperature gradually increases to 500-1200 DEG C from room temperature, calcining is kept for 30-120 min at the highest calcining temperature, and then the flexible metallic oxide nanofiber membrane is obtained. The preparation method of the flexible metallic oxide nanofiber phosphorylated peptide enrichment material is simple in process, and phosphorylated protein and phosphorylated peptide fragments canbe effectively enriched and purified efficiently and repeatedly.

The invention relates to the field of material analysis chemistry and organic chemistry. The invention provides a phosphopeptide enriching material and a preparation method and application thereof. The phosphopeptide enriching material comprises a substrate and a double-ingredient copolymer layer formed on the surface of the substrate, wherein the thickness of the double-ingredient copolymer layer is 10 to 80nm. The surface initiation-atom transfer free radical polyreaction mechanism is utilized; the double-ingredient copolymers are grafted on the surface of the substrate. The material and column solid phase extraction mode or dispersion solid phase extraction mode are organically combined; the enriching with high selectivity, high repetitiveness and high flux of phosphopeptide in a complicated mixture can be realized, so that the selective enriching of the multi-phosphopeptide and mono-phosphopeptide can be realized; the phosphorylated protein identification number can be obviously improved. Therefore the phosphopeptide enriching material is hopeful to be widely applied to the aspects of phosphopeptide enriching, further large-scale separation of phosphorylated protein and the like.

The invention discloses a phosphorylated peptide adsorption enrichment method comprising (1) a non specific adsorption inhibitor and part of a to-be-tested sample are added a sample loading buffer, and an enrichment filter column is used for phosphorylated adsorption enrichment; (2) enrichment products are collected, the abundance of 20 kinds of amino acids in non phosphorylated peptide can be calculated as first abundance, the abundance of the non phosphorylated peptide in the 20 kinds of amino acids can be calculated as second abundance, at least one amino acid with higher ratio of first abundance to second abundance can be selected as an additive; (3) the sample loading buffer added with the additive is used for phosphorylated adsorption enrichment of the to-be-tested sample. By use of the competitive adsorption effects of amino acids and peptides, adsorption of the non phosphorylated peptide can be reduced, adsorption selectivity of the phosphorylated peptide enrichment can be increased, and the method lays the foundation for improving the accuracy and stability of phosphorylation detection.

The invention relates to an immobilized metal ion affinity chromatography (IMAC) functionalized hybrid monolithic material with a high specific surface area, prepared through a sol-gel technology, andan application thereof in phosphorylated peptide enrichment. A hybrid monolithic material containing phosphate functional groups on the surface is prepared through a sol-gel reaction by taking tetraethoxysilane (TEOS) and diethylphosphatoethyltriethoxysilane (DPTS) as precursors, the hybrid monolithic material is hydrolyzed in concentrated hydrochloric acid (HCl) to generate phosphoric acid functional groups on the surface, and the hydrolyzed hybrid monolithic material is chelated with Ti<4+> to obtain the Ti<4+>-IMAC hybrid monolithic material. The Ti<4+>-IMAC hybrid monolithic material canadsorb micromolecular pyridoxal 5-phosphate under the action of metal chelation, and can also be used for carrying out high-sensitivity and high-selectivity enrichment on phosphorylated peptide fragments in beta-casein and human cervical carcinoma cell (HeLa) enzymatic hydrolysate.

The invention discloses a sequential separation and mass spectrum identification method of multi-site phosphorylation peptide. By using spinel and inverse-spinel type magnetic nano ferrite materials as a separation substrate, the method is used for realizing the fast separation from a complex sample background under the action of an external magnetic field by using the difference of the coordination property between metal ions on an octahedron binding site in the materials and different phosphorylation peptides and the inherent magnetism of the materials. The nano ferrite materials related by the invention can be used for separating a complex polypeptide mixture into non-phosphorylation peptide, mono-phosphorylation peptide and multi-site phosphorylation peptide, so that the signal inhibiting between the molecules of different samples is reduced or eliminated, in-vivo or in-source mono-phosphorylation peptide can be distinguished, and high signal-to-noise ratio and low interference are realized. The method is simple and can be used for effectively enriching low-abundance phosphorylation peptides without using a complex apparatus and realizing the sequential separation and the mass spectrum identification of the multi-site phosphorylation peptide; and the sample analysis and operation are simple without complex sample pre-treatment.

Embodiments of the present invention are directed to articles of manufacture, devices, methods and apparatus for performing liquid chromatography featuring a chromatographic sorbent having one or more pentafluorophenyl groups, wherein said one or more pentafluorophenyl groups are a bonded phase on a sorbent selected from the group comprising silica, organic polymers or hybrid organic diene material and said pentafluorophenyl groups are in a mono-, bi-, and tridentate forms.

The invention relates to peptides including DEDE(SSD)nDEG indicated by SEQ NO. 1, RRRDEDE(SSD)nDEG indicated by SEQ No. 2, RRRGDEDE(SSD)nDEG indicated by SEQ No. 3, and LKKLKKLDEDE(SSD)nDEG indicated by SEQ NO. 4, wherein n is an integer from 2 to 20. The invention also relates to phosphorylating these peptides at multiple amino acid sites by employing casein kinases. These phosphorylated peptides may be used in various applications such as forming mineralized collagen fibrils and biomimetic composites for use in tissue repair and regeneration.

Liposomes containing tau peptides, preferably phosphorylated tau peptides, and conjugates containing tau peptides, preferably phosphorylated tau peptides, conjugated to an immunogenic carrier are described. Pharmaceutical compositions and uses of the liposomes and / or conjugates for treating or preventing a neurodegenerative disease or disorder, such as Alzheimer's Disease, are also described.

The invention relates to high-selectivity enrichment of phosphorylated peptide, in particular to a method for preparing polymer materials for enriching phosphorylated peptide fragments. The method adopts monomers provided with phosphate groups to perform polymerization, so as to directly form the polymer materials provided with the phosphate groups. The materials are acted by a solution containing zirconium ions to generate phosphate ester zirconium groups, so that the materials can specially enrich and purify the phosphorylated peptide from a complex proteolysis solution. The method adopts the polymerization method to directly synthesize the polymer materials provided with functional groups, does not need to additionally perform chemical modification, and has the characteristics of simplicity and convenience.