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23 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.
Nanometer antibody and application thereof
ActiveCN119371520AOther chemical processesBiological material analysisAntiendomysial antibodiesComplementarity determining region
The invention provides a nano antibody and application thereof, a variable region in an amino acid sequence of the nano antibody comprises a complementarity determining region CDR and a framework region FR, the complementarity determining region CDR comprises a complementarity determining region CDR1, a complementarity determining region CDR2 and a complementarity determining region CDR3, the CDR3 is RWPDXY family, and binding sites of the CDR3 and AAV capsid are W99, D101 and Y103. The nano antibody provided by the invention has very high affinity and activity, and can specifically recognize and combine various serotypes of AAV, and an adsorbent prepared from the nano antibody has very strong adsorption capacity on AAV, can be applied to AAV affinity chromatography, and is beneficial to industrial application of an AAV affinity chromatography column. In addition, the method can also be applied to the field of AAV detection, and can detect empty capsid and virions together.
Owner:CROWN MEDICAL TECH DALIAN CO LTD
Novel nano material for enriching phosphorylated peptide in biological sample as well as preparation method and application of novel nano material
PendingCN119371705APreparing sample for investigationMaterial analysis by electric/magnetic meansDiseaseMedicine
The invention relates to the technical field of analysis and testing, in particular to a novel nano material for enriching phosphorylated peptide in a biological sample as well as a preparation method and application of the novel nano material. The novel nano material is an immobilized metal affinity chromatography material based on a bionic honeycomb chitosan membrane, is a biomacromolecule with good biocompatibility, has good hydrophilicity and strong metal chelating ability, and can be used for enriching low-abundance phosphorylated peptide in a biological sample in a high-selectivity manner; the result of the method is beneficial to revealing pathological mechanisms of some serious diseases and realizing early diagnosis. The analysis material provided by the invention is easy to produce and manufacture, the experimental method is relatively simple, and the analysis material can be applied to the fields of clinical medicine, life science and the like; the novel nano material disclosed by the invention has high sensitivity, high selectivity and excellent recycling stability and acid-base stability.
Owner:CHANGZHOU HUABANYA BIOTECHNOLOGY CO LTD
Purification method and application of recombinant human blood coagulation factor VII
ActiveCN119372186APeptide/protein ingredientsPeptide preparation methodsCoagulation Factor GeneRecombinant coagulation factor VIIa
The invention discloses a purification method and application of a recombinant human blood coagulation factor VII. The purification method comprises the following steps: adding a His tag into a blood coagulation factor VII gene sequence, then sequentially performing cell culture, clarification treatment, ultrafiltration concentration, nickel column affinity chromatography and anion chromatography, and finally performing self-activation to obtain the recombinant human blood coagulation factor VII. Wherein in the nickel column affinity chromatography process, the chromatography sample loading concentration is 500-1000 [mu] g / ml, and the equilibrium liquid comprises 0.5-1.0 mol / L of NaCl. According to the purification method, the purity of the recombinant blood coagulation factor VII is improved, the uniformity of the recombinant blood coagulation factor VII is higher, the specific activity of the recombinant blood coagulation factor VII is 975 IU / mg, the recombinant blood coagulation factor VII contains few other protein components, and adverse reactions caused by other components can be reduced; the cost in the production process of the recombinant blood coagulation factor VII is reduced, a specific monoclonal antibody does not need to be used, and the process cost is reduced; the production process steps of the recombinant blood coagulation factor VII are reduced, and the recovery rate is increased.
Owner:SHENZHEN WEIGUANG BIOLOGICAL PROD
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
HBC-SpyTag003 fusion gene, its expression vector, monoclonal antibody and its application
ActiveCN115976076BImmunoglobulins against bacteriaMicroorganism based processesAntigenAntiendomysial antibodies
The present invention discloses a HBC‑SpyTag003 The fusion gene, its expression vector, monoclonal antibody and its application are intended to solve the current technical problems of complex and difficult purification of SpyTag protein. The present invention uses genetic engineering technology to display SpyTag003 at high density on the surface of HBV core antigen (HBc), express it in Escherichia coli and self-assemble to form nanoparticles, immunize mice with it, and screen for high-affinity SpyTag003 monoclonal antibodies by hybridoma method; the monoclonal antibody is covalently fixed on AminoLink TM The coupled resin can be used to prepare an immunoaffinity chromatography column for purifying SpyTag protein, which has high purification efficiency and high protein purity, and can effectively retain the assembly function of the purified protein, providing a new approach for the efficient purification of SpyTag protein.
Owner:LONGHU LAB +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
Serum exosome removing device
The utility model relates to the technical field of biomedical engineering, in particular to a serum exosome removing device which comprises a centrifugal barrel and a sample injection pump, a sample injection pipe is fixedly installed at the lower end of the sample injection pump in a penetrating mode, a filtering assembly is fixedly connected to the left portion of the upper end of the sample injection pipe in a penetrating mode, and a conveying pipe is fixedly installed at the upper end of the sample injection pump in a penetrating mode. The right portion of the lower end of the conveying pipe penetrates through the upper end of the centrifugal barrel and extends into the centrifugal barrel, a centrifugal device is installed in the middle of the upper end of the centrifugal barrel in a penetrating mode, and a collecting assembly is fixedly connected to the right portion of the lower end of the centrifugal barrel in a penetrating mode. According to the serum exosome removal device, through preliminary filtration of the ultrafiltration membrane, centrifugal treatment of the centrifugal device and specific capture of the immunoaffinity chromatography column, efficient separation of serum exosomes can be achieved, the purity and accuracy of separation are improved, the whole process is relatively mild, and the operation is simple. The damage to the structure and the function of the serum exosome is reduced, and the biological activity of the exosome is favorably kept.
Owner:SHANGHAI XINZEMU BIOTECHNOLOGY CO LTD
A nano antibody against human IgG1-CH1 and a method for purifying IgG1 antibody or its fragments using the nano antibody
ActiveCN118878667BSpeed up the application processhigh puritySolid sorbent liquid separationPeptide preparation methodsFc domainProtein G
The present invention belongs to the field of biomedicine, and specifically relates to a method for capturing a nano antibody containing a human IgG1-CH1 domain, and separating and purifying an IgG1 antibody, Fab or F(ab')2 using the nano antibody. Because VHH is a single-domain antibody that is very stable, it is expected to have very low immunogenicity compared to bacterial surface proteins such as protein A and protein G, and is independent of the IgG-Fc domain to capture the antibody, which can effectively solve the problem of light chain dropout. The antibody can specifically recognize the IgG1-CH1 domain with high affinity. The affinity chromatography medium with the anti-human IgG1-CH1 domain nano antibody of the present invention as a ligand can achieve the purification of IgG1 and its Fab or F(ab')2 fragments, and has a very broad application prospect.
Owner:BIOINTRON (JIANGSU) BIOLOGICAL INC
A method for in situ enzymatic cleavage and purification of liraglutide fusion protein
The invention discloses a method for in-situ enzyme digestion and purification of a liraglutide fusion protein. The method comprises the following steps: fixing a liraglutide fusion protein containing a His tag by a nickel affinity chromatography method, completing in-situ enzyme digestion of the liraglutide fusion protein on a nickel affinity chromatography column, and realizing separation of the fusion protein containing the His tag and a liraglutide main chain in one step; recovering enterokinase by GST affinity chromatography, and further improving the purity by reverse phase chromatography to obtain the liraglutide main chain. The product recovery rate is improved by 30.26%, the product purity is improved by 6.18%, the production cost is saved, the purification efficiency is improved, and economic value is created for enterprises.
Owner:HANPEPTIDE BIOPHARMACEUTICAL GRP CO LTD
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
Semi-quantitative evaluation method for substitution degree of non-protein amino acid
The invention discloses a semi-quantitative evaluation method for non-protein amino acid substitution degree in the technical field of biological detection, which comprises the following steps: S1, expressing recombinant plasmids, and synthesizing non-protein amino acid engineered protein to be detected; s2, purifying the engineered to-be-detected protein: centrifugally collecting host cells, cracking, and purifying by using metal affinity chromatography to obtain the high-purity to-be-detected protein; step S3, preparing an LC / MS-MS sample; and S4, carrying out LC / MS-MS semi-quantitative analysis evaluation. The method can quickly, simply and conveniently evaluate the substitution degree of the non-protein amino acid in the protein, is not limited by the protein source, the protein relative molecular weight and the chemical structure and property of the non-protein amino acid, and is basically suitable for all proteins and non-protein amino acids; the technical bottleneck of non-protein amino acid substitution degree evaluation in macromolecular protein is broken through, and the blank of research technologies in related fields of protein engineering is filled.
Owner:ZUNYI MEDICAL UNIVERSITY
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
Method for high-load and large-scale separation and purification of recombinant protein A
PendingCN119552229Aincrease flow rateIncreased dynamic loadPeptide preparation methodsDepsipeptidesEscherichia coliProtein loading
The invention discloses a method for separating and purifying recombinant protein A with high loading capacity on a large scale, and provides a method for separating and purifying the recombinant protein A by using nickel ion chelation chromatography and anion exchange chromatography, the loading capacity of a nickel ion chelation chromatography sample is 40g / L, and the loading capacity of an anion exchange chromatography sample is 80g / L. According to the method, endotoxin and escherichia coli host protein are effectively removed, the purity of a recombinant protein A solution obtained through separation and purification is larger than 98% through HPLC and SDS-PAGE electrophoretic analysis, the endotoxin is smaller than 1 EU / mg, and the recovery rate is larger than 85%. The protein loading capacity is high, the technological process is simple, the cost is low, the applicability is wide, the purity and the recovery rate are high, meanwhile, the endotoxin control effect is remarkable, and therefore the protein A can be used as a raw material of immunoaffinity chromatography filler and is suitable for industrial production of recombinant protein A.
Owner:SUZHOU SEPAX TECHNOLOGIES INC
High-efficiency expression of human epidermal growth factor fusion protein and preparation method and application thereof
Owner:GUILIN MEDICAL UNIVERSITY
Method for purifying botulinum toxin
The present invention relates to a method for purifying Botulinum Toxin (BTX), and more particularly, to a method for purifying Botulinum Toxin (BTX), which comprises the steps of: purifying Botulinum Toxin (BTX) by means of a column chromatography comprising the steps of: sequentially using cation exchange chromatography (Cation Chromatography, Cation Chromatography, Cation Chromatography, Cation Chromatography, Cation Chromatography); the method comprises the following steps: (1) carrying out CEX (Hydrophobic Interaction Chromatography), (2), (3), (4), (5), (6), (7), (8) and (8); according to the present invention, the botulinum toxin having excellent purity and activity is purified by a process comprising a step of purifying a botulinum toxin by using trypsin (Trypsin), a step of treating the botulinum toxin with trypsin (Trypsin) to activate the botulinum toxin, and a step of purifying the botulinum toxin by using trypsin (Trypsin), such that the botulinum toxin can be usefully used for producing the botulinum toxin.
Owner:JETEMA CO LTD
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
Method for preparing circular RNA and nucleic acid sequence for method
PCT designated stage expiredWO2025011502A1Organic active ingredientsNervous disorderRNA PrecursorsNucleic acid sequencing
Provided is a method for preparing a circular RNA, which method comprises inserting purification tags, such as poly(A) tags or functional variants thereof into two terminal arms of a linear RNA precursor, and separating the circular RNA, which is obtained by means of the cyclization reaction of the linear RNA precursor, from other byproducts of the cyclization reaction as impurities using an affinity chromatographic medium capable of simultaneously binding to the tags. Further provided are a linear RNA precursor with a purification tag inserted in a specific region and an encoding DNA thereof. The insertion of the purification tag in the specific region does not affect the yield of linear RNA precursors produced by means of the in-vitro transcription (IVT) of the encoding DNA.
Owner:RINUAGENE BIOTECHNOLOGY CO LTD +1
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