133 results about "Protein immobilization" patented technology

Disclosed are compositions and methods for detecting small quantities of analytes such as proteins and peptides. The method involves associating a primer with an analyte and subsequently using the primer to mediate rolling circle replication of a circular DNA molecule. Amplification of the DNA circle is dependent on the presence of the primer. Thus, the disclosed method produces an amplified signal, via rolling circle amplification, from any analyte of interest. The amplified DNA remains associated with the analyte, via the primer, and so allows spatial detection of the analyte. The disclosed method can be used to detect and analyze proteins and peptides. Multiple proteins can be analyzed using microarrays to which the various proteins are immobilized. A rolling circle replication primer is then associated with the various proteins using a conjugate of the primer and a molecule that specifically binds the proteins to be detectable. Rolling circle replication from the primers results in production of a large amount of DNA at the sites in the array where the proteins are immobilized. The amplified DNA serves as a readily detectable signal for the proteins. The disclosed method can also be used to compare the proteins expressed in two or more different samples. The information generated is analogous to the type of information gathered in nucleic acid expression profiles. The disclosed method allows sensitive and accurate detection and quantitation of proteins expressed in any cell or tissue.

A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.

The present invention is to provide calixcrown derivatives of formulae 1 to 3 requisite for the preparation of a self-assembled monolayer as well as a process for the preparation thereof. Further the present invention is to provide a self-assembled monolayer which is produced by immersing a gold substrate or related metal substrate in an organic solution containing said calixcrown derivatives of formulae 1 to 3. Still further the present invention provides a process for fixing a protein monolayer by fixing proteins having molecular weight of not less than 20,000D (20KD) on said self-assembled monolayer.

The invention discloses a chip preparation method, a DNA or protein immobilization method, and a chip. The chip preparation method of the invention comprises performing weak passivation treatment after immobilization treatment, wherein the weak passivation treatment comprises contacting a weak passivation reaction solution containing a catalyst with a chip subjected to immobilization treatment, soas to promote binding of DNA or protein to the surface of a substrate, and to enable the DNA or protein to be sufficiently immobilized on the surface of the substrate. The preparation method of the invention increases the weak passivation treatment step after the immobilization treatment, so that the DNA or protein can be more fully immobilized on the surface of the substrate, the quality and efficiency of DNA or protein immobilization are improved, also DNA or protein is more fully immobilized, the relationship between the amount of the DNA or protein immobilized on the chip and the concentration of the DNA or protein initially added is more closely, and thus the immobilization amount of the DNA or protein is highly controllable, the repeatability is good, and a foundation is laid for preparing high-quality DNA or protein chips.

The invention discloses an enzyme-GO-MOFs nano composite catalyst and a preparation method thereof. The preparation method comprises the following steps: (1) adding a modifier into an enzyme liquid so as to obtain a modified enzyme solution, wherein the modifier is an N-vinyl amide polymer or pluronic F-127; the N-vinyl amide polymer is one or a mixture of more of polyvinylpyrrolidone, poly N-vinyl caprolactam and poly N-ethylene formamide; (2) mixing GO and a metallic salt solution, performing ultrasonic treatment, further adding an organic ligand solution into the mixed solution so as to obtain a GO-MOFs composite material synthesized stock solution, and performing ultrasonic treatment for another time; (3) adding a modified enzyme solution into the GO-MOFs composite material synthesized stock solution, and performing ultrasonic treatment so as to obtain a reaction liquid; (4) centrifuging the reaction liquid, collecting precipitate, washing the precipitate, and performing vacuum drying, thereby obtaining the enzyme-GO-MOFs nano composite catalyst. The preparation method disclosed by the invention is simple and feasible, and the nano composite material catalyst is gentle in synthesis condition and high in protein immobilization rate.

Disclosed are compositions and methods for detecting small quantities of analytes such as proteins and peptides. The method involves associating a primer with an analyte and subsequently using the primer to mediate rolling circle replication of a circular DNA molecule. Amplification of the DNA circle is dependent on the presence of the primer. Thus, the disclosed method produces an amplified signal, via rolling circle amplification, from any analyte of interest. The amplified DNA remains associated with the analyte, via the primer, and so allows spatial detection of the analyte. The disclosed method can be used to detect and analyze proteins and peptides. Multiple proteins can be analyzed using microarrays to which the various proteins are immobilized. A rolling circle replication primer is then associated with the various proteins using a conjugate of the primer and a molecule that specifically binds the proteins to be detectable. Rolling circle replication from the primers results in production of a large amount of DNA at the sites in the array where the proteins are immobilized. The DNA produced by rolling circle replication can be further amplified in secondary and higher order amplification processes using second-stage or higher order primers in conjunction with second-stage or higher order amplification target circles. The amplified DNA serves as a readily detectable signal for the proteins. The disclosed method can also be used to compare the proteins expressed in two or more different samples. The information generated is analogous to the type of information gathered in nucleic acid expression profiles. The disclosed method allows sensitive and accurate detection and quantitation of proteins expressed in any cell or tissue.

A high throughput screening method is described which employs a PVDF substrate for protein immobilization.

The present invention relates to a method for immobilizing a protein in a sample, which could not easily be immobilized by the conventional immobilization method, to a solid-phase; a method for quantitative determination of protein wherein an effect of inhibitory substance coexisting in a sample prepared using the immobilization method can be reduced; and a rapid and highly precise method for detecting an abnormal PrP and a method for determining BSE using the immobilization method as compared with the conventional method. The present invention provides: “a method for immobilizing a protein to a solid-phase comprising contacting the protein with the solid-phase having hydrophobic surface in the presence of a lower alcohol, and a halogenocarboxylic acid and / or a long chain alkyl sulfate, and an immobilizing reagent solution to be used therefor; a method for quantitative determination of protein comprising contacting a protein-staining solution with the solid-phase immobilized with a protein by the immobilization method, and determining a degree of color development generated thereby; an immunoblotting method wherein the solid-phase immobilized with a protein by the immobilization method is used; and a method for detecting an abnormal PrP a method for determining BSE by using the immobilization method.”

The invention discloses a living polymerization preparation method for a polyacrylic acid ball brush and application of the polyacrylic acid ball brush and particularly discloses the polyacrylic acid ball brush prepared by the living polymerization method and the application of the polyacrylic acid ball brush in the protein high density immobilization. The method comprises steps of synthesizing a novel silylanized reversible addition-fragmentation chain transfer (RAFT) polymeric chain transfer reagent; fixing the RAFT polymeric chain transfer reagent to a spherical solid phase carrier surface through a reaction; forming a polyacrylic acid brush structure on the carrier surface which is grafted with the RAFT polymeric chain transfer reagent through the RAFT polymerization; and obtaining the polyacrylic acid ball brush. The polyacrylic acid ball brush material is subjected to activation and protein immobilization, the compound of the ball brush and the protein is obtained, ant the application in the protein high density immobilization is achieved. By the aid of the method, the synthetic process and the synthetic time of the polyacrylic acid ball brush are shortened, the reaction efficiency is improved, and the polyacrylic acid ball brush has high grafting density and carboxyl content and provides more positions for protein immobilization.

A method for preparing chitosan-coated magnetic nanoparticles for protein immobilization includes forming ferrous ferric oxide (Fe3O4) nanoparticles by co-precipitation and coating the nanoparticles with chitosan in the presence of glutaraldehyde. The Fe3O4 nanoparticles can be coated by dispersing ferrous ferric oxide (Fe3O4) nanoparticles into a solution comprising chitosan and acetic acid, adding a surfactant, adding excess 50% glutaraldehyde solution, and washing the nanoparticles with a solvent. The chitosan coated ferric oxide (Fe3O4) nanoparticles can be used to immobilize proteins or other biomolecules.

The present invention provides a fibrous protein-immobilization system composition comprising a fiber comprising fiber-forming material, and a protein attached to the fiber-forming material.

It is intended to provide a novel method of immobilizing a protein and a protein chip, by which the protein can be immobilized at a high reproducibility while preventing the protein from inactivation without resort to a large-scaled apparatus and the protein can be immobilized even in a microchannel. Further, by using a cell adhesive protein as the protein to be immobilized, it is also possible to use a cell as a target and to provide a method of immobilizing a cell and a cell chip, by which a cell can be immobilized in an arbitrary region on a substrate.

To provide a measuring probe excellent in sensitivity and reproducibility, simple operation in the protein immobilization onto a support can prevent peeling or damage. A layer containing a protein is adsorbed onto a support surface to fully encircle the long axis of the support, and the adsorbed layer is treated for cross-linking with a cross-linking agent in a concentration of 1 to 5 moles to 1 mole of the protein.

The present invention relates to a method for preparing an protein monolayer using a peptide hybrid for protein immobilization, more precisely a peptide hybrid for protein immobilization which has improved solubility by introducing a PEG linker and a proper reaction group to the oligopeptide having specific affinity to selected types of proteins and is designed to provide enough space between solid substrates and proteins immobilized, whereby various solid substrates treated by the hybrid catch specific proteins effectively on. The peptide hybrid for protein immobilization of the present invention facilitates the control of orientation of an antibody on various solid surfaces and immobilization of various antibodies of different origins or having different isotypes with different affinity. Therefore, the surface treatment technique using the peptide hybrid of the invention can be effectively used for the production of various immunosensors and immune chips.

The invention relates to the field of covalently attaching proteins to a substrate, particularly to methods of immobilizing proteins by posttranslationally modifying a cysteine residue of said protein through the addition of functional groups. The invention also relates to biological molecules used in such techniques, including proteins, and detection methods and kits that utilize such immobilized proteins, such as a microdevice or “protein chip”, a high-throughput screening device, and for the microscopy of proteins on a surface.

An immobilized protein catalyst is prepared by applying an adhesive to a polymeric support, applying a layer of a globular protein over the layer of adhesive, binding a crosslinking agent to the protein layer, and binding the protein catalyst by reaction with the crosslinking agent.

Provided is a biomaterial immobilizing method including immobilizing a bovine serum albumin on a substrate, providing a biomaterial on the substrate immobilized with the bovine serum albumin, and irradiating an ultraviolet light onto the substrate provided with the bovine serum albumin and the biomaterial to immobilize the biomaterial selectively on the substrate immobilized with the bovine serum albumin.

A method for immobilizing glycoprotein is disclosed. A surface is provided. Next, a boronic acid is contacted to the surface. The boronic acid is represented as Y—R—B—(OH)2 and includes a boronic acid group(B—(OH)2), a linker R, and a functional group Y, in which the boronic acid is bonded to the surface via the functional group Y. A glycoprotein is contacted to the boronic acid, wherein two spatially adjacent hydroxyl groups of a carbohydrate chain form a boronate ester with the boronic acid group of the boronic acid to immobilize the glycoprotein. The goal of site-specific and covalent protein immobilization without interfering the binding affinity of an antibody to antigen may be thus achieved with the reaction of boronic acid and the carbohydrate chain of Fc region.

The invention discloses a method for quickly improving biological adhesiveness of a laser patterned PDMS surface. A microstructure can be formed on the PDMS surface after laser action, so that a good environment is provided for cell adhesion and growth; or the PDMS surface after laser action is chemically modified or structurally modified, so that biological adhesiveness is improved; the microstructure can fix protein, cell growth factors, enzyme and nucleic acid; a laser device generates high-energy pulse laser beams which focus to the PDMS surface through a convex lens after going through a reflection mirror, and under action of high energy and impact force caused by radiation of the high-energy pulse laser beams, the PDMS surface occurs photothermal action and photochemical action and is instantly vaporized or splashes so as to form the microstructure on the PDMS surface. Laser action is utilized for material surface modification or forming the surface microstructure, modification step of the PDMS surface is simplified without having influence on a non-modified area surface, and the PDMS surface is patterned, so that preparation efficiency is improved greatly.

The present invention provides a fibrous protein-immobilization system composition comprising a fiber comprising fiber-forming material, and a protein attached to the fiber-forming material.

The invention provides a method for fixing a protein on the surface of polydimethylsiloxane. The method is characterized in that a biotinylated polydimethylsiloxane film is prepared on the solidified surface of polydimethylsiloxane, a protein is fixed on the surface of polydimethylsiloxane by chemical technologies of biotin-avidin and water-soluble carbodiimide and a technology of 3-mercaptopropionic acid stabilized gold nano-particles, and beta-D-dodecyl-N-maltose and bovine serum albumin are utilized respectively for blockading the non-specific adsorption of a protein on the surface of polydimethylsiloxane and activated residual carboxylic groups on the surface of a 3-mercaptopropionic acid stabilized gold nano-particle. In the invention, the surface of a 3-mercaptopropionic acid stabilized gold nano-particle contains multiple carboxylic groups which can combine molecules of multiple proteins needing to be fixed thus multiple proteins needing to be fixed can be fixed on the surface of polydimethylsiloxane and a sensitivity of a subsequent enzyme-linked immuno sorbent assay (ELISA) is improved effectively.

It is intended to provide an anti-photocrosslinking group antibody capable of specifically binding to a photocrosslinking group and available in the binding of a substrate and a substance of interest in a microstructure, a complex protein including at least the antibody or at least a portion thereof, and a technique for use thereof in the detection of a target substance. The present invention provides a protein having at least the structure of an antibody that recognizes a photocrosslinking group.

A plastic substrate used for preparing biochip is a plastic carrier chip, to which the protein, peptide, or micromolecular compound is fixed after its surface is chemically treated, especially a polystyrene carrier chip, to which the oligonucleotide or polynucleotide is fixed after its surface is treated to obtain DNA chip.

The invention relates to an immobilization method of cubilose water-soluble protein. The method comprises the following steps: soaking dry cubilose in ultrapure water; dehydrating the soaked cubilose, and steaming the dehydrated cubilose in a steaming box; cooling the steamed cubilose, and the cubilose water-soluble protein is immobilized after cooling. According to the immobilization method of the cubilose water-soluble protein, ultrapure water is adopted for soaking, the ultrapure water can be easily absorbed by the dry cubilose, the cubilose is easy to soak and is not easy to dissolve; a specific steaming temperature and a specific steaming time are adopted, so that the protein is comprehensively immobilized, and the protein can not be immobilized excessively. According to the method, the cooling temperature and time are controlled, rapid cooling is guaranteed, freshness is locked in time, when the temperature is reduced to 0-4 DEG C, the fresh-keeping temperature can be kept, and the storage time can be prolonged. By using the method provided by the invention, the cubilose water-soluble protein can be immobilized, the immobilization of protein is moderate, when the cubilose is cooked, the hardness is moderate, the eating and taste are not affected, and it is good for human absorption.

The invention discloses a quaternization cellulose bead as well as a preparation method and application thereof. The diameter of the quaternization cellulose bead is 0.5-6mm; the water content of the quaternization cellulose bead is 90-99%; the framework density of the quaternization cellulose bead is 1.1-1.8g / cm<3>; the porosity of the quaternization cellulose bead is 90-99%. The quaternization cellulose bead is prepared by using a homogeneous phase modification method, and the application field of the quaternization cellulose bead is greatly widened by virtue of functional characteristics of quaternary ammonium groups, for example, as substances with negative electricity can be sucked through positive electricity of the quaternization cellulose bead, protein immobilization and the like can be achieved by virtue of affinity of the quaternization cellulose bead. The quaternization cellulose bead disclosed by the invention is good in adsorption property upon dyes and antibiotics with negative electricity, and can be used as a novel chemical material for fields such as biological carriers, chromatograph and adsorption.

The present invention provides a method for inexpensively, easily and efficiently modifying the C-terminus of a protein or peptide; a method for easily and reliably isolating a C-terminal peptide fragment of a protein or peptide; and a method for rapidly, accurately and reliably determining an amino acid sequence of a protein or peptide by using a mass spectroscope. A method comprising the step of adding a formylation reagent and a catalyst to a protein or peptide to convert a carboxyl group into an aldehyde group. A method comprising the step of reacting a nucleophilic reagent with the aldehyde group to modify the C-terminus of the protein or peptide. A method comprising the step of reacting a support having a nucleophilic group to immobilize the protein or peptide. A method comprising the step of fragmenting the immobilized protein or peptide, washing the support, and isolating the C-terminal peptide fragment from the support. A method comprising the step of subjecting the isolated C-terminal peptide fragment to mass spectrometry and determining an amino acid sequence.

The process of fixing protein or other biological molecule onto CaCO3 to prepare biosensor as nanometer inorganic carrier includes compounding colloid solution with CaCO3 and re-distilled water, adding biological molecule into the colloid solution to form mixed solution, dropping the mixed solution onto the surface of substrate electrode, evaporating 80-95 % of the solvent, and cross-linking with glutaraldehyde vapor to prepare the nanometer CaCO3-biological molecule composite biological film as the said biosensor. Thus prepared biosensor has high stability, high sensitivity, high repeatability and low needed biological molecule amount, and may be use widely in medicine, food, environment detection and other fields.

The invention discloses a separation method of a chiral medicament by applying protein functionalized magnetic nano-particles. The method comprises the following steps: (1) modifying the surfaces of superparamagnetic particles by adopting a physical adsorption method; (2) loading protein with chiral identification effect; and (3) mixing protein-modified superparamagnetic nano-particle aqueous solution and a chiral medicament racemic aqueous solution at a volume ratio of 1:1, adsorbing, and performing magnetic separation, wherein the supernatant is liquid containing the separation product of one enantiomer, and the superparamagnetic particle adsorbate is the separation product of the other enantiomer. The method is simple to operate, and has mild condition, short reaction time, and large protein immobilization capacity; the medicament identification capability of the protein is maintained; and the optical purity of the product is remarkably increased.

The invention provides a surface polylysine modifying method for a PDMS micro-fluidic chip.The PDMS micro-fluidic chip is subjected to plasma processing and silanization processing, and then coupled with polylysine, polylysine is firmly bound to the surface of the chip in a covalent binding mode, and the micro-fluidic chip capable of efficiently fixing protein is formed.According to the modified chip, the common advantages of being high in signal-to-noise ratio, high in binding capacity, low in inter-point variation coefficient and the like of a polylysine modifying method are kept, and more importantly, the problem that polylysine and the chip are bound infirmly in a physical absorption mode, and consequently polylysine is prone to fall off especially in microfluidics application is solved.