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12 results about "Affinity chromatography" patented technology
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Affinity chromatography is a method of separating biochemical mixture based on a highly specific interaction between antigen and antibody, enzyme and substrate, receptor and ligand, or protein and nucleic acid. It is a type of chromatographic laboratory technique used for purifying biological molecules within a mixture by exploiting molecular properties. Protein could be eluted by ligand solution Biological macromolecules, such as enzymes and other proteins, interact with other molecules with high specificity through several different types of bonds and interaction. Such interactions include hydrogen bonding, ionic interaction, disulfide bridges, hydrophobic interaction, and more. The high selectivity of affinity chromatography is caused by allowing the desired molecule to interact with the stationary phase and be bound within the column in order to be separated from the undesired material which will not interact and elute first. The molecules no longer needed are first washed away with a buffer while the desired proteins are let go in the presence of the eluting solvent (of higher salt concentration). This process creates a competitive interaction between the desired protein and the immobilized stationary molecules, which eventually lets the now highly purified proteins be released.
A beta-1,4-xylanase mutant and a method for constructing the same
The application discloses a beta-1,4-xylanase mutant and a construction method thereof, and comprises the following steps: constructing a beta-1,4-xylanase recombinant expression vector; introducing the recombinant vector pNZ8048-P5-XynA into a competent lactococcus lactis by using an electrotransformation technology to construct a wild-type expression vector; on the basis, designing and constructing a multi-site mutant recombinant plasmid by using a site-directed mutation method, wherein the eighth asparagine (Asn) of the beta-1,4-xylanase is mutated into tyrosine (Tyr), the 22nd serine (Ser) is mutated into proline (Pro), and the 59th alanine (Ala) is mutated into lysine (Lys); introducing the multi-site mutant recombinant plasmid into the competent lactococcus lactis to construct a heterologous expression strain of the mutant beta-1,4-xylanase (Mut-1); and stably obtaining the mutant xylanase beta-1,4-xylanase (Mut-1) with high activity by using a metal affinity chromatography (Ni 2+ ) method. The specific enzyme activity of the mutant reaches 1929.30±5.36 U / mg, and the optimal temperature is increased by 15 DEG C compared with that of the wild type.
Owner:HEFEI UNIV OF TECH +1
Recombinant uricase mutant from arthrobacter globiforme as well as purification method and application of recombinant uricase mutant
PendingCN121896187APeptide/protein ingredientsSkeletal disorderColiform bacilliArthrobacter globiformis
The invention discloses a recombinant uricase mutant derived from arthrobacter globiforme as well as a purification method and application thereof, and belongs to the field of biological medicines. The mutant is mutated at 32-34 sites, 161 sites and 184 sites of an N end of recombinant uricase derived from arthrobacter globiforme, and the mutated amino acid sequence is as shown in SEQ ID NO: 1. On the basis of obtained uricase fermentation thalli, uricase is obtained through thalli splitting decomposition, centrifugation, nickel column affinity chromatography, liquid exchange, anion chromatography and liquid exchange, in the purification method, target protein uricase is enriched through nickel column affinity chromatography, then the purity of target protein can be increased to 95% or above through anion chromatography, and the purity of the target protein can be increased to 95% or above through anion chromatography. And finally, carrying out SUMO enzyme digestion to obtain the tag-free protein. The chromatography process is used for preparing uricase in an escherichia coli recombinant expression and purification mode, and the purity and yield of uricase are greatly improved.
Owner:AFFILIATED HOSPITAL OF NANTONG UNIV
Preparation of functional ethylene receptor etr1 dimer and method for detecting activity thereof and application
This invention discloses a method for preparing functional plant ethylene receptor ETR1 dimers, along with its activity detection method and applications. The preparation method includes expressing tagged ethylene receptor ETR1 protein in a Pichia pastoris expression system, extracting ETR1 protein from membrane fractions, performing two-step affinity chromatography purification, and enriching with size exclusion chromatography to obtain biologically active ETR1 dimers. The activity detection method involves pre-incubating different concentrations of ETR1 dimers with copper ions and / or ethylene, reacting with ATP, and finally assessing its ligand-dependent kinase activity by measuring the luminescent signal associated with the remaining ATP. The advantage of this invention is that it achieves the first controllable preparation of functional ethylene receptor ETR1 dimers in a eukaryotic system and establishes a sensitive, strictly dimer-state-dependent, and copper / ethylene synergistic in vitro detection system. This system can be used for precise assessment of ETR1 protein functional sites and for high-throughput screening of potential ethylene receptor agonists or antagonists.
Owner:SOUTH CHINA UNIV OF TECH
A method for preparing equine f(ab')2 formulations using an immobilized dual enzyme platform and the preparation thereof
Owner:JIANGXI INST OF BIOLOGICAL PRODS
A sjhnf4 polypeptide antibody, preparation method and application
ActiveCN116284333BBiological material analysisImmunoglobulins against cell receptors/antigens/surface-determinantsAdjuvantWestern blot
The application discloses a SjHNF4 polypeptide antibody, a preparation method and application. The preparation method comprises the following steps: downloading a SjHNF4 protein sequence, screening a polypeptide sequence, adding a cysteine at an N terminal, then performing polypeptide synthesis and purification, coupling to obtain a recombinant polypeptide, mixing the coupled polypeptide with Freund's complete adjuvant to obtain a mixture, immunizing a mammal by using the mixture, taking heart blood of the mammal and separating serum to obtain a polypeptide antibody. The polypeptide antibody is detected by using an ELISA method, and then the polypeptide antibody is purified by using a polypeptide affinity chromatographic column. Finally, the binding affinity of the antibody and the antigen is detected by Western Blot and immunofluorescence experiments. The polypeptide antibody obtained by the application has high purity (not less than 90%) and high titer (not less than 512000), and has good purity and immunogenicity, can satisfy various experimental requirements and has commercial value.
Owner:CENT SOUTH UNIV
Purification method of lumbrukinase suitable for injection
The invention belongs to the technical field of biological pharmacy, and particularly relates to a purification method of lumbrukinase suitable for an injection. Affinity chromatography and ion exchange chromatography are adopted, and impurities such as viruses and 45k brown lumbrukinase are removed; the lumbrukinase obtained by purification is identified as proteolytic enzyme, has dual pharmacological effects of thrombolysis and fibrinogen reduction, is free of adverse reactions such as viruses, heat sources and allergy, can be prepared into powder injection and water injection, and can be used as a medicine for treating thrombolysis and fibrinogen reduction. The pharmaceutical composition can be used for thrombolysis treatment of acute myocardial infarction and acute cerebral infarction in a large dose of 1,000,000-2,400 million units, and can also be used for defibrination treatment of acute cerebral infarction in a small dose of 25,000 units. In addition, raw materials are rich in source, low in price and suitable for industrial production.
Owner:BEIJING RUZHAN BIOCHEMICAL PHARMACY RES CENT
High-efficiency expression of human epidermal growth factor fusion protein and preparation method and application thereof
Owner:GUILIN MEDICAL UNIVERSITY
A system for the production of soluble Trypanosoma brucei gambiense phospholipase A2 (TbgPLA2)
ActiveDE202025103787U1Antibody mimetics/scaffoldsHydrolasesTrypanosoma brucei gambienseInclusion bodies
A system for the production of soluble Trypanosoma Brucei gambiense phospholipase A2 (TbgPLA2), consisting of: a bioinformatics analysis unit configured to predict the 3D structure, topology, and signal peptide positions of TbgPLA2; a gene construct manufacturing unit configured to produce codon-optimized TbgPLA2 gene constructs with removed signal peptide and transmembrane sequences; an expression unit configured to express the TbgPLA2 constructs in host cells selected from the group consisting of E. coli and Pichia Pfarrer; a purification unit configured to purify TbgPLA2 from soluble fractions and inclusion bodies using nickel column metal affinity chromatography; and a refolding unit configured to renature TbgPLA2 using optimized buffer conditions consisting of Tris 55 mM, NaCl 21 mM, KCl 0.88 mM at pH 8.2.
Owner:ADEPOJU OLUWAFEMI ABIODUN +2
A method of removing interfering components from a liquid sample prior to dispensing the liquid sample on a chemical reagent test slide
A method of removing components of a liquid sample that can interfere with a test performed on a test assay using a chemical analyzer includes the steps of adding the liquid sample into a sample cup, transferring a volume of the liquid sample into a mixing cup containing an IMAC (immobilized metal affinity chromatography) resin containing porous beads to form a sample / resin solution in the mixing cup, repeatedly aspirating the sample / resin solution into and expelling the sample / resin solution from a disposable pipette tip of a pipettor of the chemical analyzer into the mixing cup to obtain a mixed sample / resin solution in the mixing cup, and allowing the mixed sample / resin solution to sit undisturbed in the mixing cup so that interfering components of the liquid sample attach to the porous beads, and the beads settle to the bottom of the mixing cup, to obtain a refined liquid sample free of the interfering components and occupying an upper portion of the mixing cup for later dispensing on the test assay.
Owner:IDEXX LABORATORIES INC
A candida albicans bivalent subunit vaccine fusion protein and a preparation method and application thereof
PendingCN122255302Aclear structureQuality is easy to controlAntimycoticsMicroorganism based processesAdjuvantPharmaceutical medicine
The application discloses a Candida albicans bivalent subunit vaccine fusion protein and a preparation method and application thereof, and relates to the technical field of biological medicines. The fusion protein is formed by connecting a Candida albicans Sap2 truncated fragment (SEQ ID NO: 1) and a Cht3 truncated fragment (SEQ ID NO: 2) through a flexible linker (SEQ ID NO: 3). The application further provides a nucleotide sequence encoding the fusion protein, a vaccine containing the fusion protein and a pharmaceutically acceptable adjuvant, and application of the fusion protein in preparation of a medicine or vaccine for preventing Candida albicans infection. The fusion protein is recombinantly expressed, enzymatically cut on a GST affinity chromatographic column and refined through ion exchange chromatography, and the purity can reach more than 95%. The fusion protein is significantly superior to a Sap2 and Cht3 physical mixing group, has an immune synergistic enhancement effect, and can be used for preventing Candida albicans infection.
Owner:LICHI BIOLOGICAL PROD (CHONGQING) CO LTD
Method for purifying follicle stimulating hormone
The present invention relates to a method for purifying follicle stimulating hormone (FSH) in high yield and high purity, said method comprising an immunoaffinity chromatography (IAC) step.
Owner:LG CHEM LTD
A universal immunoaffinity column for modular component spytag protein, its preparation method and purification method
ActiveCN115970338BImmunoglobulins against bacteriaSolid sorbent liquid separationEscherichia coliAntigen
The application discloses a universal immunization affinity column for modular component SpyTag protein, a preparation method and a purification method, and aims to solve the technical problems of complex and difficult SpyTag protein purification. The application realizes high-density display of SpyTag003 on the surface of HBV core antigen (HBc) through genetic engineering technology, expresses and self-assembles the SpyTag003 into a nanoparticle in Escherichia coli, immunizes mice, and obtains a high-affinity SpyTag003 monoclonal antibody through a hybridoma method; the monoclonal antibody is covalently fixed on an AminoLink TM The coupling resin is used for preparing the immunization affinity column, the immunization affinity column can be used for efficiently purifying proteins with SpyTag labels at N-termini or C-termini from prokaryotic or eukaryotic expression systems, the column is simple to prepare, can be effectively regenerated, and has simple and conventional elution conditions, is low in cost, is high in purification efficiency, is high in protein purity, and can effectively reserve the assembly function of the purified protein, and provides a universal new method for efficient purification of SpyTag proteins.
Owner:LONGHU LAB +1