66 results about "Bifunctional reagent" patented technology

The invention provides a new method for double quantifying the protein group marked on the basis of stable oxygen isotope <18>O, which is characterized in that: based on the fact that the N end peptide section and the C end peptide section are generally labeled, and by means of combining the <18>O chemical labeling and enzyme digestion, the quantifying strategy for two <18>O markings of the same sample can be accomplished. The method relates to the reductive alkylation and enzyme digestion of protein, dual-functional reagent modifying and <18>O marking of peptides, multi-dimensional liquid phase chromatogram separation and multistage Tandem Mass Spectrometry quantification and identification. A first mass spectrum peak area is used to quantify; and the identification of the protein is conducted through a second stage mass spectrum and by searching the database. The invention also provides a reagent box for facilitating the implementation of the method.

The invention provides a magnetic nano-particle immobilized basic protease, the magnetic nano-particle immobilized basic protease is obtained by utilizing a silylating reagent of 3-aminopropyl-triethoxysilane to modify Fe3O4 nano-particles so as to improve water solubility and stability of the Fe3O4 nano-particles, introducing a free amino terminal onto the surface of the Fe3O4 nano-particles, and then using a bifunctional reagent of p-phenylene diisothiocyanate to fix the basic protease into the Fe3O4 nano-particles modified by the silylating reagent, and the magnetic nano-particles with surface functionalization are taken as a vector for immobilizing the basic protease so as to be applied in green transformation of biomass. The preparation method is simple and easy to operate, and a biological catalyst can be rapidly and conveniently recycled through an externally applied magnetic field, thereby being conductive to realization of continuous large-scale production through biological catalytic reaction.

The invention provides a method for preparing an enzyme labelling antibody. The method comprises the following steps of: concentrating a colloidal gold solution, and then adjusting the pH value of the concentrated colloidal gold solution to be slightly greater than the pH value of an isoelectric point of an enzyme for labelling; adding an aqueous solution of the enzyme for labelling into the colloidal gold solution, mixing the solutions, and then connecting the antibody to molecules of the enzyme on the surface of colloidal gold by a bifunctional reagent; and carrying out centrifugation to obtain an enzyme labelling colloid gold compound antibody. In the method, enzyme labelling on the colloidal gold is accomplished in a direct-absorption simple mode; the separation of the enzyme and a coupling agent and separation purification after connection of the antibody can be finished only by the simple centrifugation without the complicated and high-loss expensive steps of gel filtration, dialysis or affinity chromatography; by concentration operation prior to enzyme absorption by the colloid gold, the connection amount of a connected reagent can be increased, and the concentration of gold colloid particles in the enzyme labelling colloid gold compound antibody can be increased simultaneously; and the enzyme labelling colloid gold compound antibody is accordant with the characteristic.

The present invention provides one kind of nanometer bacterial magnetic corpuscle carrying primary amino group medicine and its preparation process. The nanometer bacterial magnetic corpuscle consists of three structural units, including homotype double-function reagent, primary amino group medicine and nanometer bacterial magnetic corpuscle. The nanometer bacterial magnetic corpuscle carrying primary amino group medicine has high medicine carrying capacity, less aggregation, easy separation and purification, anticancer and antitumor effect similar to that of pure primary amino group medicine, but much lower toxicity.

The invention belongs to the field of biological catalysis, and particularly relates to a novel immobilized enzyme preparation, and a preparation method and application thereof. The preparation method of the novel immobilized enzyme preparation comprises the following steps: combining nano magnetic chitosan microspheres and a metal-organic framework material having so many properties such as multiple pores, large specific area, multiple metal sites and the like through graft copolymerization to obtain a nano magnetic chitosan-metal organic framework material having a core-shell structure with a porous network structure; and mixing the nano magnetic chitosan-metal organic framework material used as a carrier and an enzyme solution, and immobilizing at 0-25 DEG C for 1-12 hours to obtain the novel immobilized enzyme preparation. The method can immobilize enzymes such as lipase, proteinase, peroxidase and the like, and has the advantages of low cost, high immobilization efficiency, favorable enzyme stability, high enzyme loading amount and high enzyme recovery rate; no bifunctional reagent is used in the immobilization process; and the method is simple to operate, low in cost and mild in reaction.

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-FGand 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 relates to a method for labeling and colorimetric detecting the biology molecule by surface fictionalized gold or silver nanometer particle. The probe molecule for identifying the biology molecule and the nail molecule for connecting the gold or silver nanometer particle first through the gluglucosan whose double function agent is covalence reformatted macromolecule to form the deformation coated the gold or silver nanometer particle, it uses the self-component on the surface of the gold or silver nanometer particle by the nail group of the deformation to form gluglucosan molecule coated gold or silver nanometer particle after purifying and quantifying the deformation.

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-FGand 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.

A process is disclosed for coating the inner surface of a solid support (e.g. a capillary or a microchip channel) to reduce the analyte adsorption to the surface for separating components in a fluid stream in contact with the surface. The process comprises (a) covalently binding a bi-functional reagent to the support surface, (b) mixing the monomer with a radical initiator under an oxygen-removed environment; and (c) rapidly bringing the above mixture solution to the functionalized surface under an oxygen-free environment.

The invention discloses a preparation method of three-dimensional nitrogen-doped graphene, which adopts a dual-function reagent thioacetamide solution process to synthesize the three-dimensional nitrogen-doped graphene (3D G-N). The method comprises the following steps: thioacetamide is decomposed into NH3 and H2S during hydrolysis, and the NH3 is used as a nitrogenous component to implement nitrogen doping of the graphene (G-N); when Zn<2+> exists in the solution, S<2-> and the Zn<2+> form a ZnS particle precipitate which is dispersed among the nitrogen-doped graphene lamellae; and dilute hydrochloric acid is utilized to dissolve out the ZnS to obtain the porous three-dimensional nitrogen-doped graphene. The preparation method is simple and environment-friendly, and can greatly widen the application of the graphene in the fields of optics, electricity, magnetics and the like.

This invention describes a first report on the synthesis of certain 124I-labelled photosensitizers related to chlorines and bacteriochlorins with long wavelength absorption in the range of 660-800 nm. In preliminary studies, these compounds show a great potential for tumor detection by positron emission tomography (PET) and treatment by photodynamic therapy (PDT). The development of tumor imaging or improved photodynamic therapy agent(s) itself represent an important step, but a dual function agent (PET imaging and PDT) provides the potential for diagnostic body scan followed by targeted therapy.

The biologically degradable packing film is prepared through enzymolyzing konjaku powder with alpha-amylase to prepare hydrosol, esterifying the hydrosol with acetic anhydride to prepare gelatin with partially enclosed hydroxy in konjaku glucomannan, adding plasticizer to prepare konjaku fluid, mixing with PVA fluid, adding double-functional reagent to prepare homogeneous crosslinked gelatin, stoving and filming in a casting machine. The packing film has certain water tolerance, acid and alkali resistance and adequate strength, and used film can be decomposed by environmental microbes into CO2 and water.

The invention discloses an amperometric enzyme electrode based on an electrostatic spinning fibrous membrane and a method for preparing the same. The method uses dimethyl formamide as solvent, a platinum electrode, a gold electrode, a glassy carbon electrode or a carbon paste electrode as a receiving electrode and acrylonitrile copolymer as raw materials, and deposits a copolymer fibrous membrane layer on the surface of the electrode through the electrostatic spinning method. The method is as follows: firstly, a monolayer of redoxase is constructed on the surface of fibers through the covalent method; secondly, bifunctional reagent is added into enzyme solution, and enzyme congeries is driven to be covalently fixed on the enzyme membrane on the surface of the electrode. In the amperometric enzyme electrode, due to large specific surface area and high porosity of the fibrous membrane and the unique enzyme immobilization method, the electrochemical enzyme electrode is characterized by large response strength, high stability, wide linear detection range and so on. The method for preparing the enzyme electrode is simple and is suitable for mass production, and the enzyme electrode can be reused and preserved for a long term.

The invention provides a preparation method of a novel polysaccharide conjugate vaccine treating a PEG heterobifunctional reagent as a conjugation bridge. A meningococcal polysaccharide conjugate vaccine is exploited through the method. The method has the following advantages: 1, the respective self-crosslinking of polysaccharides and proteins in traditional methods is avoided, the yield is improved, and the quality control is benefited; and 2, a long PEG chain can increase the distance between the polysaccharides and the proteins, reduce the mutual space shield effect between the polysaccharides and the proteins, and improve the immunogenicity of the polysaccharide conjugate vaccine.

This invention discloses a protein chip, its preparation method and its usage. The chip is composed of processed carriers and small molecule protein conjugates, among which, the carrier surface is processed by aldehyde group reagent, pesticide and papaverine anti-body molecular are fixed on the carrier with a covalent bond by double-function crosslinker. This invented protein chip is prepared by preparing small molecule protein conjugates, aminating the carrier glass carrying chip, crosslinking double function reagenrt, fixing and packaging anti-body, useing in testing residuals of pesticide and papaverine.

The invention relates to a degradable packaging film comprising basic components of: 5-85% of konjac fine powder, 0.01-0.1% of alpha-amylase, 2-10% of acetic anhydride, 5-85% of polyvinyl alcohol, 0.01%-0.05% of an aldehyde bifunctional reagent, and 1-10% of glycerol. The manufacturing method comprises the steps that: konjac fine powder is subjected to enzymolysis by using alpha-amylase, such that a hydrosol is obtained; the hydrosol is esterified by using acetic anhydride, such that a gum in which partial hydroxyl in konjac glucomannan of the konjac fine powder is sealed is prepared; a plasticizing agent is added, such that a konjac fine powder gum-solution is prepared; the konjac fine powder gum-solution and a polyvinyl alcohol gum-solution are blended; the bifunctional reagent is adopted, such that a homogeneous-phase cross-linked gum is prepared; the gum is dried on a casting machine, such that the required packaging film is obtained. The packaging film provided by the invention has certain water resistance, acid and alkali resistances, and enough strength. When the film is discarded, the film can be completely decomposed into carbon dioxide and water by microbes in the environment.

The invention relates to a method for preparing an amino-functionalized micron gold and anion exchange resin composite material and belongs to the technical field of biological chemical separation. The method comprises the following steps of: firstly, absorbing water-soluble chlorine and gold anions to an anion exchange resin surface; secondly, restoring by using a chemical reducing agent to prepare a micron gold and anion exchange resin composite material; and finally, modifying the surfaces of gold micron particles by using an amino sulfydryl bifunctional reagent to obtain the amino-functionalized micron gold and anion exchange resin composite material. By the method, the amino-functionalized micron gold and anion exchange resin composite material can be efficiently obtained, and the amino-functionalized micron gold and anion exchange resin composite material can be applied to extraction of short-carbon chain volatile organic mixed acids in fermentation liquid.

The invention discloses a method for immobilizing fructosyl transferase by taking mesoporous molecular sieve-chitosan as a carrier, and belongs to the technical field of food enzyme engineering. The method comprises the following steps of: absorbing zymoprotein on a mesoporous molecular sieve SBA-15 which is taken as a carrier, adding chitosan and a bifunctional reagent glutaraldehyde and furtherabsorbing and cross-linking, and immobilizing the fructosyl transferase by a two-step absorption-crosslinking method so as to firmly immobilize enzyme molecules on the carrier. The recovery rate of enzyme activity of the immobilized enzyme is over 75 percent, and the thermal stability and the pH stability are superior to those of free enzyme. Compared with other immobilization methods, the invention has the advantages that: the method is easy to operate and lower in cost, and has a good immobilization effect, conditions are mild, and the enzyme molecules are difficult to leak; and the product, namely the immobilized fructosyl transferase can be applied to the production of high-purity fructooligosaccharides.

The invention relates to a derivatization and separation analysis method of amino pyrazolone iso-bifunctional reagents of reducible carbohydrate chains and glycoprotein O-carbohydrate chains. The method particularly includes the steps: performing derivatization on the reducible carbohydrate chains and the amino pyrazolone iso-bifunctional reagents by two reaction modes; performing reductive amination to generate carbohydrate chain derivatives with active methylene groups under acidic conditions; performing Michael addition reaction to generate carbohydrate chain derivatives with primary aminegroups under alkaline conditions. The two derivatives generated by the derivatization method have different active groups, the carbohydrate chains can further be analyzed and researched, and the derivatization method is high in derivatization efficiency and high in universality. The invention further provides releasing and marking method of the amino pyrazolone iso-bifunctional reagents for glycoprotein O-carbohydrate chains under alkaline conditions. According to the method, O-carbohydrate chain derivatives with primary amine groups are acquired, and sample preparation processes are greatly simplified.

The present invention discloses a preparation method for a novel red blood cell substitute-artificial red blood cell fluorescent nanoparticles. According to the present invention, a cross-linking technology is adopted to carry out a cross-linking reaction for hemoglobin (Hb), catalase (CAT), superoxide dismutase (SOD) and carbonic anhydrase (CA) to generate a polyHb-SOD-CAT-CA complex body, wherein the cross-linking technology is performed by a bifunctional reagent glutaraldehyde; the polyHb-SOD-CAT-CA complex body is further embedded by polylactic acid and polyethylene glycol copolymer (PLA-PEG) nanoparticles. With the present invention, the antioxidase of the SOD and the CAT in the complex body can eliminate the oxygen free radicals generated during the blood transfusion process, such that the ischemia-reperfusion injury easily generated during the blood replacement process can be avoided; with the CA, the red blood cell substitute of the present invention has oxygen carrying performance and carbon dioxide transportation performance; with the PLA-PEG microcapsule embedding, the half life period of the artificial blood product can be prolonged, the biocompatibility of the artificial blood product can be increased.

The present invention discloses a modified epoxy resin immobilized enzyme, and a preparation method and an application thereof. The preparation method comprises the following steps: modifying epoxy resin, adding polyethyleneimine to the modified epoxy resin for further modification, and then adding an enzyme to be immobilized and glutaraldehyde for immobilization to obtain the modified epoxy resinimmobilized enzyme. By applying the technical scheme, the epoxy resin is modified, the polyethyleneimine is added to the modified epoxy resin for further modification, aldehyde groups in the resin are covalently combined with amino groups in the polyethyleneimine to bind various enzymes, then the bifunctional reagent glutaraldehyde is used for activation, by using the method, space resin arms areincreased to form a network structure, the enzyme can be more easily bound by covalent bonding, and enzyme load can also be improved due to reduced space inhibition.

The invention relates to a method for detecting causal agent and left drug in whole animal blood, wherein it uses gene engineering method or chemical couple method to couple the causal agent antigen, causal agent antibody, drug antibody with single chain antibody or single colon antibody of red blood cell to obtain one dual-function agent which can combine red blood cell and combine causal agent antigen, causal agent antibody, and left drug antibody. The inventive agent can use the red blood cell of animal as instructor to detect the causal agent and left drug of animal.

This invention provides to rare earth element metal chelation label protein for quantitative protein study method and agent case. This invention method relates to protein restore alkali base and pancreatic enzymes slices, peptide section double function agent decoration, rare metal chelation, multi-diemension liquid phase spectrum isolation and multiple degrees series spectrum quantitative and testing. Through one degree spectrum ion signal intensity content and protein validating through series mass spectrum and database. This invention also describes this method agent case.

The invention provides an immobilization method of xylanase and immobilized xylanase. In the invention, xylanase is immobilized on the surface of a 3-aminopropyltriethoxysilane-modified Fe3O4@SiO2 nanoparticle carrier by a covalent bond (peptide bond) through the binding function of a bifunctional reagent to obtain immobilized xylanase, which limits the spatial configuration transformation of xylanase, so as to realize the heat tolerance, acid and alkali resistance, and resistance to interference of metal ions and inhibitors of xylanase; and the immobilized xylanase has relatively high tolerance to heat (40-70 DEG C), relatively wide adaptability to extreme pH values (3.0-8.0), relatively high tolerance to metal ions (Ba<2+>, Fe<2+>, Mn<2+>, Ca<2+>) and interfering agents (blue moon hand sanitizer, Triton X-100, Tween 20, EDTA solution, ethanol), and excellent operational stability. The activity of the immobilized xylanase can still be maintained at 73% or higher after being repeatedly used for 9 times.

The method is provided for characterizing agglomeration of particles in a liquid containing an analyte, including introducing liquid into a fluid chamber; mixing the liquid with a bifunctional reagent, lighting the fluid chamber using an excitation light beam extending through the fluid chamber in a longitudinal direction (X); acquiring at least one image using a matrix photodetector, each image including pixels (In(x,y), x,y representing the coordinates of a pixel of an image, the image (I(x,y)) being formed by radiation transmitted by the lighted fluid chamber; and calculating from at least one acquired image (I(x,y)), at least one indicator (Ind2) characterizing the particle agglomeration. The photodetector can be positioned less than 1 cm from the fluid chamber, and during the calculation step, the calculated indicator (Ind2) is representative of the intensity of the pixels of the image. The method advantageously allows for charactering the agglomeration of particles in a liquid.

The invention relates to a modification method of polymmeric hemoglobin, it is characterized by: preparing purified hemoglobin from natural red blood cells in the condition of 0-4DEG C and nitrogen, first polymerizing them through same type bifunctional reagent, then modifying them with 5'-pyridoxal phosphate. During the modifying process, glutathione should be added after adding PLP 0.5-1.5 hours. The arts can control generation of ferrihemoglobin, but also ensuret the P50 value in appropriate range and reduce production costs greatly.

A polysialic acid compound is reacted with a hetero-bifunctional reagent to introduce a pendant functional group for site-specific conjugation to sulfhydryl groups, for instance side chains of cysteine units in drugs, drug delivery systems, proteins or peptides. The functional group is, for instance, an N-maleimide group.

The present invention relates to a dual function reagent composed of a polymeric chain and end caps, transfer fibers made from cellulose fibers and the dual function reagent, a liquid transfer layer made from the transfer fibers, and absorbent articles incorporating the liquid transfer layer. Embodiments of the present invention also relate to methods of making and using the dual function reagent, transfer fibers, transfer layer and absorbent articles.

The invention discloses a preparation method of a cross-linked beta-lactamase aggregate for treating pharmaceutical wastewater. The preparation method comprises the following steps: transferring a recombinant beta-lactamase gene sequence into a host cell E.coli Top10 to obtain recombined bacteria; carrying out induced expression, ultrasonic crushing and centrifugal treatment and taking supernatant to obtain rough enzyme liquid; adding bovine serum albumin into the rough enzyme liquid as an aggregation agent and a protection agent; precipitating and aggregating to form an enzyme aggregate; crossly linking the obtained enzyme aggregate through a bidirectional reagent so as to prepare the cross-linked beta-lactamase aggregate. After a test, the cross-linked beta-lactamase aggregate prepared by the invention is high in enzyme activity and good in stability, can be repeatedly utilized and is used for treating antibiotic wastewater; the chemical oxygen demand is reduced by 72.9 percent; the cross-linked beta-lactamase aggregate has an important application value in the treatment of the antibiotic wastewater in a pharmaceutical industry.

This invention describes a first report on the synthesis of certain 124I-labelled photosensitizers related to chlorines and bacteriochlorins with long wavelength absorption in the range of 660-800 nm. In preliminary studies, these compounds show a great potential for tumor detection by positron emission tomography (PET) and treatment by photodynamic therapy (PDT). The development of tumor imaging or improved photodynamic therapy agent(s) itself represent an important step, but a dual function agent (PET imaging and PDT) provides the potential for diagnostic body scan followed by targeted therapy.