109 results about "Affinity chromatography" patented technology

The invention overcomes the limitations described for the bonding of structured layers by providing a method for selectively reducing the bonding of materials. In its most generic form, the invention uses a bonding technique in combination with a printing method for modifying or covering at least one portion of a surface to either fully or partially prevent localised bonding. The structuring process may act upon the layers either before or after the bonding of the layers. The invention overcomes the limitations described in the application of affinity chromatography by providing a planar substrate with discrete optical detection flow cells that contain porous material and have connecting microchannels for fluid delivery and/or removal, and a method for making the same.

The present invention relates to a method of separating one or more immunoglobulin containing proteins from a liquid. The method includes first contacting the liquid with a separation matrix comprising ligands immobilised to a support; allowing the immunoglobulin containing proteins to adsorb to the matrix by interaction with the ligands; followed by an optional step of washing the adsorbed immunoglobulin containing proteins; and recovering said immunoglobulin containing proteins by contacting the matrix with an eluent which releases the proteins. The method improves upon previous separation methods being that each of the ligands consists essentially of a monomer or dimer SpA or protein Z or a functional variant thereof.

The invention relates to a construction method and an application of high-yield gamma-aminobutyric acid recombinant escherichia coli/pET-28a-1pgad, in particular to a genetic engineering bacterium construction method, recombinant enzyme enzymology property study and an application to the conversion of L-glutamic acid for producing gamma amino butyric acid (GABA), which belongs to the technical field of biology in the fermentation engineering. Firstly, lactobacillus plantarum GB 01-21 glutamic acid decarboxylase (GAD) genes are obtained through polymerase chain reaction (PCR) amplification, recombinant plasmids pET-28a-1pgad are constructed, in addition, the successful expression is realized in E.coli BL21(ED3), secondly, Ni column affiliation chromatography purification is adopted on crude enzyme liquid for obtaining recombinant GAD, in addition, the enzymology property of the recombinant GAD is primarily studied for guiding the optimization of conversion conditions, finally, conversion experiments are carried out on a 5L fermentation tank, the GABA accumulation concentration can reach 204.5g/L, the mol conversion rate is 97.92 percent, and good foundation is made on the further industrial application.

The invention relates to methods for isolating a product and/or reducing turbidity and/or impurities from a load fluid comprising the product and one or more impurities by passing the load fluid through a medium, followed by at least one wash solution comprising arginine, and collecting the product using an elution solution. The invention further relates to a product prepared using a method as described herein.

The invention discloses fusion protein with anti-tumor, anti-inflammation and oculopathy-treatment functions and a preparation method and application thereof, which belong to the technical field of biological pharmacy. Specifically, the invention relates to the fusion protein of an integrin blocking agent, which has the function of inhibiting tumor angiogenesis and has the integrin affinity and the combining capacity. A rigid (R) or flexible (F) linker is adopted to fuse two polypeptides, so as to respectively obtain protein A and protein G, the medicine effect can be increased, the half-life period can be extended, the stability can be enhanced, and the fusion protein has the characteristics of strong action effect, low toxicity and the like and can be used for preventing and treating solid tumors, all kinds of inflammation and angiogenesis oculopathy. The fusion protein structurally comprises two angiogenesis inhibiting polypeptides and the amino acid linker between the two angiogenesis inhibiting polypeptides, and the fusion protein is expressed in colibacillus or eukaryocyte by using a genetic engineering method and is obtained through separation and purification of a GST (Glutathione S Transferase) affinity column.

A method for preparing immune nanogold test paper used to quickly detect furazolidone includes synthesizing coupled matter of flurazolidone ¿C cattle seralbumin and coupled matter of flurazolidone ¿C catalase ; using coupled matter of flurazolidone ¿C cattle seralbumin as immune total antigen; making immune experiment on rabbit to obtain multiple clone antibody of rabbit resistance furazolidone; preparing nanogold reagent being used to label said antibody; fixing IgG antibody and coupled matter of furazolidone ¿C catalase on nitrocellulose film; sealing unreacted active group; using above ¿C prepared film, antibody and paper as well as sample pad to form said test paper.

The invention discloses expression and preparation methods for a bivalent bispecific antibody and a hybrid protein and belongs to the technical field of biology. According to the methods, the aim of preparing products, i.e., the bivalent bispecific antibody and the immune hybrid protein thereof are prepared through expressing all parts of the bivalent bispecific antibody and the immune hybrid protein thereof separately in appropriate procaryotic or eucaryotic cell systems, carrying out high-performance affinity-chromatography purification and separation, and then, carrying out splicing in vitro under the condition of intein splicing. The methods have high production efficiency, are wide in applicable application range and facilitate the separation and purification of the products. The products prepared by using the methods have biological activities, i.e., bivalent bispecific immunoaffinity and cytotoxicity. The methods are novel methods for preparing biological drugs for directional attack on cancers or other diseases.

The invention discloses a purified rHSA-IFN alpha2b producing method of pichia fermentation liquor from expressing and restructuring albuminar-interferon alpha2b anastomosing protein, which comprise the following steps: fermenting and expressing high density pichia engineering bacteria of protein gene by integrated albuminar-interferon alpha2b anastomosing; combinating and purifying sequentially fermentation liquor with rHSA-IFN alpha2b with affinity column chromatography, hydrophobic column chromatography, ion exchange column chromatography and gel column chromatography; Getting high-purity medicinal recombination albuminar- interferon alpha2b anastomosing protein by large scale preparing. The invention operates simply and convenience.

The invention relates to the field of biotechnology. A vascular endothelial cell grow factor (VEGF) plays an important function in the regeneration and hyperplasia of blood vessels; but the VEGF in the body has low content and a short half life; therefore, sufficient endogenous VEGF protein is hardly extracted to meet the ever growing experimental study and clinical requirements. The invention discloses a method for economically and conveniently preparing, extracting, purifying and recombining the VEGF protein of a human body through a recombination gene engineering and immunity affinity column method. The invention also discloses a method for carrying out the in-vitro non-radioactive chemical coupling label processing of the purified VEGF protein. The VEGF protein (including the protein having no label or having the chemical coupling label) can be widely applied to experimental study and clinical diagnosis and treatment.

A dual-antibody sandwith ELISA detecting method for human soluble p185HER-2 antigen features that the purified and coated P185HER-2 monoclonal antibody is bound to solid supporter, the specimen containing soluble p185HER-2, the purified standard p185HER-2 antigen and the coated antibody are incubated, and another purified and coated p185HER-2 monoclonal antibody marked by horseradish perioxidase is used as detecting antibody. It can be used for serum diagnosis of tumor.

The invention aims at providing a novel chitin enzyme with a high-efficiency degradation rate. The high-activity soluble chitin enzyme is obtained through cloning a gene, which is used for coding thechitin enzyme, in a genome of streptomyces albolongus ATCC (American Type Culture Collection) 27414, constructing a recombinant vector and transferring the recombinant vector into escherichia coli BL21 (DE3) to carry out inducible expression. Through Ni column affinity chromatography, the chitin pure enzyme is purified and obtained; the enzymatic property and hydrolysis mechanism of the chitin pure enzyme are researched; the optimum temperature and optimum pH (potential of Hydrogen) of the enzyme are 55 DEG C and 5.0 respectively; the specific enzyme activity is 66.2U/mg; a main product of degraded chitin is chitobiose, and the chitin enzyme is expected to be applied to the preparation of a chitooligosaccharide, and has potential application value.

The invention discloses a method for finding a protein interaction with baculovirus PCNA (Proliferating Cell Nuclear Antigen). The method is characterized by comprising the following steps of cloning full-length DNA of an autographa californica nuclear polyhedrosis virus (AcMNPV) PCNA gene by utilizing the means such as molecular cloning, and DNA sequencing, purifying the full-length DNA, coupling the full-length DNA to an affinity column, and fining the interaction protein by adopting the mass spectrography. The method can be used for finding and functionally studying interaction protein of the baculovirus PCNA.

The invention relates to a preparation method of a chromatography stationary phase for chitosan oligosaccharide separation. The invention aims to solve the problems of a small number of chromatographic columns for chitosan oligosaccharide separation, unsatisfactory separation effect, and difficult acquisition of high purity single-component chitosan oligosaccharide. The method consists of: 1. preparing activated silica gel; 2. preparing chloro silica gel; 3. preparing benzene boric acidification silica gel, i.e. the benzene boric acidification affinity chromatography stationary phase for chitosan oligosaccharide separation. The invention provides the separation method of benzene boric acidification affinity chromatography stationary phase for chitosan oligosaccharide separation.

This invention discloses a method for enriching and purifying glycosylated protein. The immobilization of agglutinin comprises: adding epoxy ethyl derived magnetic particles into a centrifugal tube, performing immobilization reaction, washing, sealing, washing repeatedly to immobilize agglutinin onto epoxy ethyl derived magnetic particles, adding buffer and storing. The separation and purification of glycosylated protein comprises: immobilizing prepared agglutinin onto epoxy ethyl derived magnetic particles, adding standard glycosylated protein for adequate coupling, washing repeatedly, eluting to obtain enriched and purified glycosylated protein, loading into a dialyzing bag, and desalting. This invention solves the problems of complex enriching and separation process, low recovery rate and high cost faced by background technology. This invention has such advantages as simple process, high recovery rate, low affinity chromatographic column cost, and convenient usage.

The present invention relates to the purification of polypeptides and the removal of endotoxin via immobilized metal affinity chromatography (IMAC). More specifically, the invention relates to methods for removing bacterial endotoxin in a protein solution. In specific embodiments, the invention relates to the elimination of endotoxin from Moraxella catarrhalis outer membrane proteins.

A method of preparing immunoglobins with high valence and high specialty against respiratory syncytial virus and adenovirus. It comprises culturing respiratory syncytial virus and adenovirus with Hep-2 cell strain, Hala Strain, preparing virus antigen, or preparing protective antigens according to sequences of respiratory syncytial virus and adenovirus by the genetic engineering technology; then,centrifuging the prepared virus antigens and getting purified antigens, or concentrating antigens by 20úÑ sucrose, or purifying antigens by chromatographic absorbing columns, screening high-valent immunoglobins against respiratory syncytial virus and adenovirus, and neutralizing virus in viro; finally extracting immunoglobins from bloods containing high-valent immunoglobins against respiratory syncytial virus and adenovirus by a low-temperature alcohol method or ion exchanging chromatography and so on.

The invention discloses a preparation method of a citrinin immunoaffinity column, belongs to the technical field of immunoaffinity chromatography and biotoxin detection, and discloses the preparation method of the citrinin immunoaffinity column. Immunogen is prepared by connecting citrinin and carrier protein through a formaldehyde condensation method. An immune animal gets an anti-citrinin antibody. Cupric embedded ionic polymer is prepared in a mass polymerization method. Immunoaffinity adsorbent which is prepared by adsorbing the polymer to the anti-citrinin antibody is filled into the citrinin immunoaffinity column which is prepared in a solid phase extraction pipe. The citrinin immunoaffinity column which is prepared in the preparation method of the citrinin immunoaffinity column is capable of combining with the citrinin in a specific mode, and cirtrinin solution with high purity and concentration is obtained through purification and enrichment. The maximum combination volume of the citrinin immunoaffinity column which is prepared in the preparation method of the citrinin immunoaffinity column is 597 ng, citrinin fortified recovery in a sample is 83.7% to 93.2%, and the recovery rate is no lower than 80% when the citrinin immunoaffinity column is repeatedly used for three times. The preparation method of the citrinin immunoaffinity column can be applied to rapid detection of the citrinin in fermented food.

The present invention relates to the use, for affinity purification of an antibody or an fragment of an antibody, of a ligand-substituted matrix comprising a support material and at least one ligand covalently bonded to the support material, the ligand being represented by formula (I)

L-(Sp)_v-Ar¹—Am—Ar²   (I)

wherein L, SP, Ar¹, AM, Ar² and v are defined herein.

The invention relates to a kit for purifying melamine residues in animal derived food such as milk, milk powder and egg. The kit comprises an immunoaffinity chromatographic column for purification of melamine, an eluent, an eluate, a regeneration liquid and a balance preservation solution. An adsorbent loaded in the immunoaffinity chromatographic column for the purification of melamine is composed of a solid carrier and a melamine polyclonal antibody coupled with the carrier; the melamine polyclonal antibody is obtained by using a conjugate of a melamine hapten with a carrier protein as an immunogen; and the melamine hapten is a melamine analogue 4, 6-diamino-1, 3, 5-triazine-2-acetic acid. The purification method provided by the invention combines with a chromatography method or a chromatography tandem mass spectrometry method for efficient detection of melamine content and has high selectivity. The method has the advantages of simple operation and good purification effect and avoids the interference of impurities on the determination results; and the immunoaffinity chromatographic column can be used repeatedly to reduce analysis cost and environmental pollution.

The invention discloses a recombinant human fibronectin III1-C (rhFNIII1-C), and a preparation method and application thereof. The preparation method comprises the following steps: optimizing a targetgene segment of the target protein according to codon expression preference, and inserting the obtained gene segment into pET-32a to construct a recombinant plasmid; transforming and introducing therecombinant plasmid into an expression vector BL21(DE3), and carrying out screening to obtain positive clone bacteria capable of realizing efficient soluble expression; carrying out enlarging fermentation culture on the positive clone bacteria, carrying out induced expression, performing crushing and centrifuging to obtain a supernatant, and carrying out purification through elution with a His-tagaffinity chromatography column, dialysis, filtration and other steps to obtain a high-quality rhFNIII 1-C solution with a protein concentration of 1.0 mg/mL or above and a purity of 90% or above. According to detection and observation results of cell adhesion promoting tests, the rhFNIII1-C has the performance of obviously promoting adherence and adhesion of MDBK cells and Balb/c/3T3 cells and rapid division and growth of the cells, which indicates that the rhFNIII1-C has huge potential of being used as a raw material and a finished product for cosmeceuticals and medical skincare.