45 results about "Protein enrichment" patented technology

The invention relates to a multifunctional double-layer core-shell structure magnetic nano particle. In the invention, a magnetic nano particle with a particle size of 1-300 nm is used as a core and coated with a double-layer shell consisting of a SiO2 layer with a thickness of 1-200 nm and a hydrolyzed silane coupling agent layer is 1-100 nm thick and comprises one or more multifunctional groups; the particle size and the shell layer thickness can be controlled through regulating the volumes, the weight ratios and the reaction time of the magnetic core, a silicon dioxide precursor, a silane coupling agent and a catalyst in a preparation process; the total particle size of the nano particle can be as small as 5-50 nm and as large as 700-800 nm; the nano particle can have superparamagnetism, paramagnetism and ferromagnetism according to the change of the magnetic core particle size; and one or more bioactive molecules can be connected into the shell layer of the magnetic nano particle or to the surface of the shell layer through a chemical method or a physical method. The invention also provides a preparation method of the multifunctional double-layer core-shell structure magnetic nano particle and application thereof. The particle preparation method has the advantages of simplicity, moderate condition, low cost and easy realization of industrial production. The nano particle can obtain different functions through connecting different bioactive molecules and can be applied to the fields of protein enrichment, biological detection, separation and purification, targeted drug carriers, cell imaging and medical imaging.

The invention discloses an integrated proteomic sample pretreatment platform based on an SCX (strong cation exchange)/SAX (strong anion exchange) mixed filling material and application thereof. The proteomic sample pretreatment platform comprises a pipette gun head (1), strong cation exchange resin and strong anion exchange resin mixed filling materials (2) and a solid phase extraction film (3), wherein the solid phase extraction film (3) is loaded in the lower end of the pipette gun head (1); the strong cation exchange resin and strong anion exchange resin mixed filling materials (2) are loaded in the lower end of the pipette gun head (1), and are positioned above the solid phase extraction film (3). The proteomic sample pretreatment platform has the advantages that the steps of sample preenrichment, reduction, alkylation and enzymolysis are completed on the mixed ion exchange filling materials; a reactor is characterized in that the used mixed ion exchange filling materials can realize the protein enrichment at any pH values; in the enzymolysis pH replacement process, the protein loss is little; trypsinase can move back and forth between the two kinds of filling materials; the enzymolysis efficiency is higher.

The invention relates to a sugar protein enrichment purification method, according to the method, hydrazide chemical method for high selective sugar protein enrichment and a centrifugal ultrafiltration device for efficient purification are integrated, hydrazide sugar protein enrichment process can be completed in the centrifugal ultrafiltration device, the whole sugar protein enrichment process can be completed in situ, the sample loss can be effectively reduced, at the same time, by use of the centrifugal ultrafiltration device, a solubilizing agent can be applied in the enrichment process so as to realize sugar protein simultaneous enrichment in soluble protein and / or insoluble membrane protein; by use of the advantages of effective solvent replacement of the centrifugal ultrafiltration device, various types of elution solutions can be introduced into the sugar protein enrichment process to complete effective removal of non sugar protein so as to enhance the selectivity of sugar protein enrichment.

The invention belongs to the technical field of glycopeptide/glycoprotein enrichment materials and analysis, and relates to a sugar chain releasable glycosylated peptide fragment/protein enrichment material based on a hydrazide strategy, a preparation method of the material and an application of the material. The surface of a matrix micro-sphere is bonded with a hydrazide compound containing disulfide bonds to form a functional material with a hydrazide group on the surface, wherein the functional material can be used for glycopeptide enrichment. The disulfide bonds are led to the surface of the material to realize sugar chain releasable enrichment of glycopeptide/glycoprotein, sugar chains are in covalent binding, and the material has the advantage of high enrichment efficiency and overcomes the shortcoming that the sugar chains are not easily released in a traditional hydrazide method and cannot be used for O-glycosylated peptide fragment/protein enrichment. The sugar chain releasable enrichment material is easy to prepare and good in enrichment selectivity, is widely applicable to N-glycosylated and O-glycosylated peptide fragment/protein enrichment and has a high practical value in the fields of glycoproteomics and the like.

The invention relates to the field of drug target protein enrichment, separation and detection, and discloses a flavone glycoside functionalized magnetic nanometer affinity probe, and further providesa preparation of the flavone glycoside functionalized magnetic nanometer affinity probe, applications of the flavone glycoside functionalized magnetic nanometer affinity probe in the drug target protein capture and/or identification of cells, and a method for capturing an intracellular target protein through the flavone glycoside functionalized magnetic nanometer affinity probe, wherein the flavone glycoside functionalized magnetic nanometer affinity probe comprises magnetic nanoparticles, a coupling structure and a flavone glycoside compound supported on the surface of the magnetic nanoparticle, one end of the coupling structure is coupled to the surface of the magnetic nanoparticles, and the other end of the coupling structure is coupled to the flavone glycoside compound. With the flavone glycoside functionalized magnetic nanometer affinity probe of the present invention, the high-specificity capture can be achieved only with a small amount of the whole protein extraction liquid.

The invention belongs to the technical field of protein enrichment, and relates to a specific reversible newborn protein enrichment method which comprises steps: marking cells with AHA, and extractingtotal proteins; taking cell total proteins and an alkynyl resin to be connected through click reaction; washing non-specifically adsorbed proteins with different buffer solutions to remove the non-specifically adsorbed proteins; carrying out enzyme digestion by trypsin to release a target peptide fragment; collecting samples for mass spectrometry. Compared with the prior art, according to the method, high-selectivity enrichment of a newly synthesized low-abundance protein is achieved through biolabeling, selective covalent bonding, strict elution conditions and selective dissociation, large-scale analysis and identification of protein can be achieved by combining nano-LC-MS/MS, and the newly-generated protein can be further identified according to the added mass label. The method providesan efficient method for efficient enrichment and accurate identification of the newborn protein, and is widely applied to the research field of low-abundance new proteomics.

The present invention relates to a novel silica gel material capable of recognizing N-phosphopeptides and proteins under neutral conditions and application thereof in N-phosphopeptide and protein enrichment. Non-porous silica microspheres are prepared by a seed growth method, a core-shell microsphere initial layer is formed by a template-guided dissolution and re-deposition method, and finally sub-2-micron core-shell silica gel with vertical pores can be obtained by acid reflux. N-tert-butyloxycarbonyl-L-tyrosine and N, N'-dipicolylamine are reacted to form a support molecule, and a phosphaterecognition functional molecule is formed by complexation of a metal ion on the support molecule. Finally, the novel phosphoric acid recognition functional silica gel material is obtained by covalently bonding the phosphoric acid recognition functional molecule with the sub-2-micron core-shell silica gel having the vertical pores through an amide bond. The novel phosphoric acid recognition functional silica gel material can rapidly and specifically enrich the N-phosphopeptides and proteins in a neutral buffer system.

Proposed is concentrated milk with the improved taste, and the concentrated milk can be obtained by: (a) separating raw milk or whole milk in a method known per se to obtain a skimmed milk fraction and a cream fraction; (b) subjecting the skimmed milk fraction obtained from step (a) to nanofiltration and obtaining a mineral-enriched permeate P1 and a partially mineral-removed and protein-enrichedretentate R1; (c) mixing the retentate R1 with at least part of the cream fraction obtained from step (a); (d) optionally dehydrating the mixture obtained from step (c) to obtain a concentrate; (e) mixing the concentrate obtained in step (d) or the mixture obtained in step (c) with a buffer containing or consisting of at least one phosphate and at least one hydroxycarboxylate to obtain a mixture;(f) homogenizing the mixture obtained in the step (e) to obtain a homogenized product; and (g) pasteurizing the homogenate obtained from step (f).

The invention discloses a pretreatment method, a preservation method, an automatic treatment system and a detection method of a urine sample, and relates to the technical field of biological detection.The pretreatment method comprises the steps that the urine sample subjected to protein splitting is subjected to reductive alkylation treatment, and then protein enrichment and enzymolysis are conducted; in the protein enrichment step, a PVDF (Polyvinylidene Fluoride) filter plate is adopted to carry out protein enrichment on the sample subjected to reductive alkylation treatment. The invention further provides an automatic urine sample treatment system and an automatic sample treatment method. The treatment system greatly reduces the labor intensity of people, facilitates the improvement of the treatment efficiency of urine sample treatment, meets the high-flux and automatic proteomics pretreatment requirements, and adapts to the reproducibility and flux of the current clinical requirements.

The invention discloses a protein enriching device. The protein enriching device mainly comprises a shell, a water drawing pump and a foam separation device, wherein a stirring device, a centrifugingdevice and a ball milling device are arranged in the shell and are used for extracting protein; the foam separation device is used for enriching the protein. Through the protein enriching device, liquid supernatant can be obtained by only adjusting a control device and a driving device to crush and centrifuge cells in raw liquid; the process is simple; the manpower and the time are saved; meanwhile, protein enriching liquid obtained by a foam separation method is excellent in effect, simple in treatment process, high in treatment capacity and low in cost and low in energy consumption.

The invention relates to a photo-induced multifunctional cross-linking agent as shown in a general formula (I), wherein the photo-induced multifunctional cross-linking agent is mainly applied to biomacromolecule interaction, such as protein-protein interaction and protein-nucleic acid interaction. According to the invention, the cross-linking agent can be used in biological samples or cells of cell lysis solutions to capture protein-protein interaction or protein-nucleic acid interaction, and can be applied to subsequent protein enrichment and protein cross-linking mass spectrometry; and the photo-induced multifunctional cross-linking agent has important application potential and practical value in interaction research of biomacromolecules.

The invention discloses puffed black fungus powder and a preparation method thereof. The preparation method comprises the following steps of cleaning black fungi, and mashing into small blocks; drying the black fungi at the drying temperature of 40 to 60DEG C for 30 to 50 minutes, and enabling the water content to be 8 to 30 percent; performing vacuum microwave puffing treatment on the black fungi, wherein the microwave power is 650 to 780W, the puffing time is 5 to 12 minutes, and the vacuum degree is 0.05 to 0.10MPa; feeding the puffed black fungi into a crusher for crushing to 100 meshes, thus obtaining the black fungus powder. The puffing rate of the black fungi is 250 to 450 percent. According to the puffed black fungus powder and the preparation method thereof disclosed by the invention, firstly, the black fungi are puffed and are then crushed, so that microstructures of the black fungi are changed, the enrichment ratio of protein as well as the dissolution rate, the solubility, the medicinal effect and the bioavailability of functional substances such as polysaccharide and the like are remarkably improved, and quick absorption and intake of multiple nutrients by a human body are facilitated.

Research of secretory proteomics has wide application in the aspects of early diagnosis of diseases and treatment of diseases. A traditional secretory protein enrichment method mostly adopts serum-free culture, but the culture mode has various defects. The invention provides a protein enrichment process for two-step peptide fragment layer enrichment, and the specificity of the method is greatly improved. Meanwhile, according to the invention, an unnatural amino acid, namely Azidohomoalanine (AHA), which can be used for replacing methionine is introduced to mark the protein synthesized by the cells per se. And in combination with a click chemical reaction which is widely applied in recent years, alkynyl modified biotin (biotin) can be efficiently added to an insertion site of AHA. With the help of biotin as a marker, separation, purification and identification of secreted protein in a serum-containing culture medium can be realized.

The invention belongs to the technical field of medicines, and particularly relates to a method for enriching sulfydryl nitroso protein in serum. According to the method, firstly, high-abundance interference proteins (albumin and immune globulin) in a serum sample are selectively removed, and then nitrosolated proteins in serum are further enriched through a specific labeling and affinity chromatography combined method. The method disclosed by the invention is good in specificity and simple to operate, and the problems of complex operation, large impurity influence and low enrichment efficiency of the existing mercaptonitrosylated protein enrichment are solved.

The invention discloses construction and application of a membraneless organelle in prokaryotic escherichia coli. According to the method, an expression vector of recombinant spider silk protein or arthropod-like elastin of protein consisting of a tandem mass of repeated peptide segment monomers of nephila cavaleriei dragline silk protein MaSp1 shown as Seq ID No. 1 and protein consisting of a tandem mass of MaSp2 repeated peptide segment monomers shown as Seq ID No. 2 is constrcuted, and is introduced into an expression host; after induced expression, a soluble protein enrichment phase, namely a membrane-free chamber, is formed; and component protein in the membrane-free area exists in a soluble state, and the biological activity of the component protein is reserved to the maximum extent.According to the method, 1,3-propane diamine is produced through the membraneless organelle in escherichia coli for the first time, and the nanoparticles are synthesized in the membraneless organellefor the first time, so that the method is milder and more environment-friendly than a chemical synthesis method.

Proposed is a concentrated milk with the improved stability, and the concentrated milk can be obtained by: (a) separating raw milk or whole milk by means of a method known per se to obtain a skimmed milk fraction and a cream fraction; (b) subjecting the skimmed milk fraction obtained from step (a) to nanofiltration and obtaining a mineral-enriched permeate P1 and a partially mineral-removed and protein-enriched retentate R1; (c) mixing the retentate R1 with at least part of the cream fraction obtained from step (a); (d) optionally dehydrating the mixture obtained from step (c) to obtain a concentrate; (e) mixing the concentrate obtained from step (d) or the mixture obtained from step (c) with at least one hydrocolloid and at least one buffer to obtain a mixture; (f) homogenizing the mixture obtained in the step (e) to obtain a homogenized product; and (g) pasteurizing the homogenate obtained from step (f).

The present invention relates to a method for protein enrichment of a biomass of red unicellular algae such as Galdieria and to the biomass thus obtained.