50 results about "Immobilized protein" patented technology

A method for the preparation of an immobilized protein ultrathin film reactor by immersing a solid support alternately into an aqueous solution of protein, and into an aqueous solution of polyion charged oppositely to said protein and preparing a structurally controlled ultrathin film of mono- or multi- protein layers on a said solid support with precision at molecular level. And a method for chemical reaction of a substrate by preparing an immobilized protein ultrathin film reactor composed of multiple layers of protein on a solid support by above mentioned method, and initiating a chemical change of the substrate molecules using obtained immobilized protein ultrathin film reactor.

The invention provides method of detecting interactions of small molecules with target molecules.

The invention belongs to the field of immobilization of proteins, and relates to preparation and application of a dopamine functional magnetic nano-carrier. The preparation comprises the following steps: firstly, dissolving FeSO4.4H2O and FeCl3.6H2O into distilled water, high-speed stirring and under protection of nitrogen, adding ammonia water of 25% NH4OH, and adding oleic acid; secondly, repeatedly cleaning black precipitates obtained by a stirring reaction and then using a strong magnet to perform precipitation separation, and drying to obtain oleic acid magnetic nano-particles; and finally, adding the oleic acid magnetic nano-particles into a dopamine hydrochloric acid solution, after the reaction is over, using a magnet to separate, cleaning and drying obtained deep brown precipitates, and then the dopamine functional magnetic nano-carrier can be obtained. According to the preparation and application of the dopamine functional magnetic nano-carrier provided by the invention, dopamine hydrochloride is adopted for modifying magnetic oleic acid nano-particles, so that the surface of a magnetic nano-particle has a plentiful of quinonyls and amidogen functional groups, thus proteins can be effectively immobilized; besides, the surface of the carrier has a layer of soft poly dopamine oxide films, the surface area is larger, and the preparation method is simple.

The present embodiments provide systems, kits and methods suitable for performing dry or substantially dry electro-blotting analyses on immobilized protein or nucleic acid samples. Electro-blotting performed according to the presently described embodiments may include a step whereby detection of one or more immobilized proteins or nucleic acids is electrophoretically accelerated. Methods for performing electro-blotting of immobilized proteins or nucleic acids may include applying an electric voltage to one or more reagents typically used in protein or nucleic acid blotting procedure. The one or more reagents may be absorbed on a suitable carrier matrix. Electro-blotting performed in accordance with the systems and methods described herein may be performed under substantially dry conditions (i.e., with little or no aqueous buffers).

The invention discloses a method for purifying and directionally immobilizing histidine tag protein and application. According to the method disclosed by the invention, tannic acid, metal ions and a hydrophilic anti-protein adhesion polymer are self-assembled and modified on the surface of a carrier to obtain a material surface which is efficiently chelated with cobalt ions, so that one-step purification and immobilization of the histidine tag protein is realized; a ligand of trivalent cobalt has inertia and divalent cobalt ions are oxidized into trivalent cobalt ions so that the stability of the immobilized protein can be improved. The method disclosed by the invention has the advantages of simple process and low price; a separation and purification or immobilization operation process for the histidine tag protein of the method is simple and effective, so that the technology has a wide application prospect.

Methods and reagents for site-selective functionalization of peptides and proteins. The methods most generally involve the reaction of a thioester with hydrazine. Reagents include bifunctional reagents of formula:

H₂N—NH—CH₂-M-L-FG

and salts thereof where M is a single bond or a chemical group carrying a non-bonding electron pair, such as —C(O)NR′—, where R′ is H, or an alkyl or aryl group; L is an optional linker group as described above; and FG is a functional group having reactivity that is orthongonal to that of the hydrazine group. FG can, among others, be an azide, alkenyl, alkynyl, nitrile (—CN) or triazole group and is preferably an azide group (—N₃). Methods and reagents can, for example, be combined with intein-mediated protein splicing to link proteins or fragments thereof to various chemical species or to a surface. Surface immobilization of proteins via the methods herein results in immobilized proteins which substantially retain biological activity and is thus useful for the generation of peptide or protein microarrays. Kits for functionalization and/or immobilization of peptides and proteins are provided as well as microarrays of peptides, proteins or both.

The invention discloses the preparation of a tin dioxide-three-dimensional graphene (SnO2-3DGR) nanocomposite material immobilized protein-modified electrode and the application research of electrochemical sensing, which belongs to the technical field of biosensors. The method provided by the present invention is to prepare SnO2‑3DGR nanomaterials, which are drop-coated on the surface of ionic liquid carbon paste electrode (CILE), and then myoglobin (Mb) is drop-coated on the surface of CILE modified by SnO2‑3DGR. After natural drying, chitosan (CTS) solution was added dropwise and dried to prepare the third-generation electrochemical enzyme sensor. The sensor prepared by the invention has the characteristics of simple process, convenient operation, low cost, high detection sensitivity, strong electrical signal and simple electrode pretreatment, and can be used to detect organic small molecule trichloroacetic acid (TCA), with a detection limit of 1.67 mmol L -1.

The present invention reates to a method for detecting activated ras protein. The method includes immobilizing a protein on a solid support, incubating the immobilized protein with lysates from cultured cells, where the lysates include activated ras protein, and determining the amount of activated ras protein bound to the immobilized protein. The present invention also relates to a method of detecting ras oncogenic related malignancy in a human subject.

The invention discloses a method for screening and characterizing a metabolite-protein interaction system based on metabolomics and a structural mass spectrometric technique; a heterologously expressed tagged target protein is immobilized through an affinity immobilization method, then, the target protein is incubated with a metabolite extract, and finally, after being separated, denatured and eluted, the target protein-metabolite complex is subjected to non-targeted metabolomics analysis, and the complex is compared with a control group (an affinity vector without immobilized protein) so as to screen a metabolite with potential interaction with the target protein. Afterwards, the potential interaction system is directly verified using structural mass spectrometry. The binding method enables high-throughput screening of metabolites with potential interaction with the target protein, and also, direct characterization of the structural mass spectrometry can be used to screen out false positive interaction results that are probably introduced during indirect characterization. The method is a reliable high-throughput research tool for protein-metabolite interactions.

The invention relates to the technical field of gene engineering, in particular to CD86-based membrane fixed protein, and a preparation method and application thereof. CD86 comprises a signal peptide encoding gene, an extracellular region encoding gene, a transmembrane region encoding gene and an intracellular region encoding gene; and the membrane fixed protein is to replace the extracellular region encoding gene of the CD86 with an expressible exogenous gene. Lentivirus-containing host cells can be used for cultivating cells, and can supply nutritional substances for the cells continuously, additional nutritional substances are not needed, and convenience and rapidness are realized.

It is an object of the present invention to provide a method of conveniently efficiently and easily immobilizing a protein on a substrate, a method of detecting a substance capable of interacting with the protein at a high sensitivity, and a protein and a protein-immobilized substrate which are used in these methods. In the method of detecting a substance capable of interacting with a protein using the immobilized protein of the present invention, a protein bound to a compound having a group capable of binding onto a substrate for immobilizing a biomolecule or a carrier provided on the substrate and immobilized onto the substrate, is used as the immobilized protein.

The present invention provides a method for detecting modified LDL, abnormal cells or bacteria using an intermolecular interaction analysis method, in which a region involved in ligand recognition by a receptor is expressed, without modification or as a biotinylated protein, in cells or in a test tube, and thereafter, the expressed region or the expressed biotinylated protein is immobilized via avidin or streptavidin to a solid phase while the orientation thereof is maintained, and the immobilized protein is utilized; and a kit for detecting the modified LDL or the like.

The invention provides methods to assess protein stability and to obtain sizing information. In one aspect, the screen comprises a 94 detergent panel and a series of MWCO filtered microplates. A protein of interest is bound to an affinity matrix and aliquoted into a 96-well microplate. Wells containing the immobilized protein are washed in the new detergent and then eluted in the new detergent into a collection plate. Protein not stable in the new detergent is precipitated on the resin and not present in the elutions. Half of the elution is passed through a high (i.e., 300 kDa) MWCO microplate and the other half through a low (i.e., 100 kDa) MWCO microplate. Elutions from the microplates are spotted on a nitrocellulose membrane, visualized by Western analysis (or by some other method), and quantified. The high MWCO provides stability readout and the ratio of low/high kDa provides sizing information.

The invention belongs to the field of protein immobilization and discloses preparation of a functionalized graphene oxide carrier and an application method of the functionalized graphene oxide carrier in rapid immobilization of proteins. According to the technical scheme, the invention has the key points of preparing and modifying functionalized graphene oxide; P-beta-sulfatoethylsulfonyl aniline (SESA) is adopted to activate the graphene oxide (GO), diazotization is carried out on the activated carrier, and finally, the carrier is used for rapidly immobilizing the proteins. The functionalized graphene oxide carrier has a lamellar structure and is large in surface area and simple in preparation method. According to the preparation method and the application provided by the invention, three different proteins in different shapes are used for immobilization, the immobilization can be rapidly finished with the functionalized graphene oxide as an immobilization carrier, and in addition, the immobilization yield and the immobilization efficiency can both reach 80-90 percent. Therefore, the method of preparing the functionalized graphene oxide carrier for immobilizing the proteins provides a new thinking to study rapid immobilization and has certain study significance.

The invention relates to the field of immobilized protein, in particular to a method for preparing heterologous protein wrapped polyhedrosis. The bombyx mori cytoplasmic polyhedrosis protein, a fusion foreign protein with a bombyx mori cytoplasmic polyhedrosis virus structural protein VP5 tag and a protease specific cleavage site are co-expressed in bombyx mori culture cells or bombyx mori by using a baculovirus expression system through a genetic engineering method, so that the polyhedrosis wrapping the foreign protein is formed. The formed polyhedron can be purified through simple differential centrifugation; the polyhedrin has protection and slow release effects on foreign proteins wrapped by the polyhedrin. After the purified polyhedrosis is cracked under an alkaline condition, the pH value is adjusted to a polyhedrosis protein isoelectric point by using a hydrochloric acid solution, the polyhedrosis protein can be precipitated through centrifugation, and the fusion foreign protein is retained in a supernatant, so that the fusion foreign protein can be quickly and conveniently obtained; the fusion foreign protein is subjected to enzyme digestion through corresponding protease to remove a VP5 tag, so that a foreign protein closer to the nature can be conveniently obtained.

The invention discloses a method for immobilizing a protein by using a biofilm. The microbial biofilm is used as an initial material, and chemical modification with a polysaccharide is performed to obtain a biofilm material for immobilization of a target object, wherein the target object comprises a free amino-containing protein and /or an amino-containing nanoparticle. The biofilm material disclosed by the invention can be used to immobilize proteins with catalytic activity or non-catalytic activity, and free amino-containing nanoparticles, and the stability of the immobilized protein can besignificantly improved. In addition, the method is universal and can be extended into the use of universal type biofilms, and the utilization range of biofilm materials is greatly improved.

The invention discloses a method for immobilizing protein by oriented affinity of a conducting polymer modified vector. According to the method disclosed by the invention, the vector is modified by a conducting polymer, and oriented immobilized protein is constructed on the basis of biological affinity between prosthetic group and apoprotein. The method disclosed by the invention has main advantages that the conductive vector is constructed by virtue of the conducting polymer; in addition, the protein is highly oriented, which is more conducive to the contact between an active center of the protein and a reaction substrate; and the protein, which is immobilized on the basis of the oriented affinity, is obviously greater than protein, which is immobilized by virtue of a normal adsorption method, in activity; therefore, a feasible method is provided for further constructing a high-efficiency and low-cost bio-catalytic system.