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131 results about "Biotinylation" patented technology
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In biochemistry, biotinylation is the process of covalently attaching biotin to a protein, nucleic acid or other molecule. Biotinylation is rapid, specific and is unlikely to disturb the natural function of the molecule due to the small size of biotin (MW = 244.31 g/mol). Biotin binds to streptavidin and avidin with an extremely high affinity, fast on-rate, and high specificity, and these interactions are exploited in many areas of biotechnology to isolate biotinylated molecules of interest. Biotin-binding to streptavidin and avidin is resistant to extremes of heat, pH and proteolysis, making capture of biotinylated molecules possible in a wide variety of environments. Also, multiple biotin molecules can be conjugated to a protein of interest, which allows binding of multiple streptavidin, avidin or neutravidin protein molecules and increases the sensitivity of detection of the protein of interest. There is a large number of biotinylation reagents available that exploit the wide range of possible labelling methods. Due to the strong affinity between biotin and streptavidin, the purification of biotinylated proteins has been a widely used approach to identify protein-protein interactions and post-translational events such as ubiquitylation in molecular biology.
Kit for detecting multiple cytokines in tumor immunity as well as preparation method and application of kit
The invention belongs to the technical field of biological detection, and provides a multiple cell factor detection kit in tumor immunity and a preparation method and application thereof, and the detection kit comprises a capture antibody, a detection antibody, a mixed protein standard substance, a diluent and a washing buffer solution. The capture antibody is prepared by coupling a biotinylated antibody with a fluorescent microsphere coated with biotin to obtain a capture antibody coupled with a fluorescent encoding microsphere, and the detection antibody is prepared by coupling derivatized phycoerythrin PE-SMCC with a specific thioether bond of a sulfhydrylated antibody to obtain a fluorescent labeled antibody; the mixed protein standard substance adopts a freeze-drying protection system containing BSA, trehalose and the like. The single hole of the kit can synchronously detect various cell factors, the sample dosage is only 25 microliters, the kit is suitable for immune monitoring of iron overload and other chronic inflammation models, compared with a traditional ELISA method, the efficiency is improved by 10 times, the cost is reduced, and the kit has the remarkable advantages of being high in throughput, wide in linear range and high in stability.
Owner:WUHAN SAIXIAOMAN BIOTECHNOLOGY CO LTD XIANNING BRANCH
PH response type probiotic-liposome hybrid system as well as preparation method and application thereof
The invention belongs to the technical field of biology, and particularly discloses a pH response type probiotic-liposome hybrid system, a preparation method and application thereof, and the pH response type probiotic-liposome hybrid system comprises engineered escherichia coli Nissle 1917, pinocembrin-loaded biotinylated liposome and an enteric polymer; wherein the engineered escherichia coli Nissle 1917 is in non-covalent binding with the biotinylated liposome loaded with the pinocembrin through surface display streptavidin, so that EcNPIN is formed, and the EcNPIN is coated with an enteric polymer, so that the pH response type probiotic-liposome hybrid system is formed. The pH response type probiotic-liposome hybrid system EcNPIN-L provided by the invention can resist the acidic environment of the stomach, completes pH response release and colonization in the intestinal tract, solves the problems of lack of intestinal tract localization of drugs and low probiotic treatment efficiency, improves the local intestinal tract concentration of pinocembrin, and promotes intestinal tract colonization of probiotics.
Owner:JIANGSU TAIZHOU PEOPLES HOSPITAL +1
Method for detecting Anti-drug antibodies against self-assembling trimeric biologics using an affinity capture elution-protein a / g assay
A method and kit are provided for detecting anti-drug antibodies (ADAs) against self-assembling trimeric biologics, such as XPro1595, a pegylated variant of soluble human Tumor Necrosis Factor, in biological samples, particularly human serum. The method employs an Affinity Capture Elution-Protein A / G (ACE-AG) assay that overcomes non-specific reagent interactions inherent in conventional bridging and standard ACE assays due to the biologic's dynamic monomer exchange. The assay utilizes a two-plate system: a streptavidin-coated plate with biotinylated trimeric biologic captures ADAs, which are eluted, neutralized, and transferred to a protein A / G-coated plate for specific immunoglobulin capture and detection with a sulfo-tagged trimeric biologic via electrochemiluminescence. The kit includes pre-coated plates, labeled biologics, elution and neutralization solutions, assay buffer, confirmatory reagent, and instructions. The assay offers a robust, specific, and sensitive solution for immunogenicity testing of trimeric biologics.
Owner:INMUNE BIO INC
Separation and analysis method for ctEVs with time resolution capability and application of separation and analysis method
PendingCN120624363ACell dissociation methodsMammal material medical ingredientsHeterologousAnimal use
The invention discloses a ctEVs separation and analysis method with time resolution capability and application thereof. The ctEVs separation and analysis method comprises the following steps: S1, culturing cells by adopting glycometabolism markers; or injecting the animal by adopting a glycometabolism marker; step S2, manufacturing a fishbone-shaped micro-fluidic chip modified by streptavidin; s3, separating and analyzing the new tumor source extracellular vesicles by the micro-fluidic chip, and co-incubating the extracellular vesicles and the aptamer probe co-modified by Biotin and DBCO to realize biotinylation of the new tumor source extracellular vesicles; the aptamer probe is a heterologous multivalent aptamer probe; step S4, capturing biotin-labeled target extracellular vesicles by adopting a chip; and S5, acquiring a chip image by using a fluorescence microscope, and analyzing the concentration of the target extracellular vesicles in the sample according to a fluorescence signal. The method can be used for screening tumor circadian rhythm markers and the like.
Owner:XIAMEN UNIV
Kit for detecting multiple cytokines in macrophage polarization as well as preparation method and application of kit
The invention belongs to the technical field of biological detection, and provides a multiple cell factor detection kit in macrophage polarization and a preparation method and application thereof, and the detection kit comprises a capture antibody, a detection antibody, a mixed protein standard substance, a diluent and a washing buffer solution. The capture system is used for preparing a capture antibody coupled with a fluorescence coding microsphere by coupling a biotinylated antibody with a fluorescence microsphere coated with biotin, and the detection system is used for preparing a fluorescence labeled antibody by coupling derivatized phycoerythrin PE-SMCC with a specific thioether bond of a sulfhydrylated antibody. The mixed protein standard substance adopts a freeze-drying protection system containing BSA, trehalose and the like. The single hole of the kit can synchronously detect multiple cell factors, the sample dosage is only 25 microliters, the kit is suitable for in-vitro tests of mouse mononuclear macrophage leukemia cell lines RAW264.7, compared with a traditional ELISA method, the efficiency is improved by 10 times, the cost is reduced, and the kit has the remarkable advantages of being high in throughput, wide in linear range and high in stability.
Owner:WUHAN SAIXIAOMAN BIOTECHNOLOGY CO LTD XIANNING BRANCH
Method for detecting hydrogen peroxide by utilizing self-biotinylation of guanine quadruplex
PCT designated stageWO2026043246A1Microbiological testing/measurementBiological testingGuanine-QuadruplexesHemin
The present invention relates to a composition, a kit and a detection method for detecting hydrogen peroxide on the basis of self-biotinylation according to the peroxidation activity of a guanine quadruplex, and, more specifically, to a composition, a kit and a detection method for detecting hydrogen peroxide, the composition comprising: i) a guanine quadruplex-hemin complex (G-quadruplex-hemin complex); ii) biotinyl tyramide; and iii) a probe capable of binding to the guanine quadruplex.
Owner:KOREA ADVANCED INST OF SCI & TECH
A single-line-state oxygen self-circulation aggregation-induced emission homogeneous microsphere and a preparation method and a no-washing detection method thereof
This invention discloses a homogeneous microsphere based on singlet oxygen self-circulating aggregation-induced emission, its preparation, and a wash-free detection method, which improves the sensitivity and efficiency of wash-free luminescent oxygen channel immunoassay. This invention successfully prepared AIE receptor microspheres by embedding the photosensitizer AIER dye, ultra-bright AIEG dye, and singlet oxygen-responsive dye SO into carboxyl-modified polystyrene nanoparticles. Simultaneously, an antibody-1-labeled AIE receptor microsphere solution, an antigen-containing test solution, and biotinylated antibody-2 were added to a 96-well plate, followed by streptavidin-modified donor microspheres. After wash-free incubation, the fluorescence signal value was measured, and the concentration of antigen in the test solution was obtained according to a standard curve. This platform completes wash-free, highly sensitive detection within 5 minutes, innovatively combining the aggregation-induced emission characteristics of AIEgens and a singlet oxygen cycling strategy, providing an efficient and universal solution for wash-free diagnostics.
Owner:SOUTH CHINA UNIV OF TECH
Crosslinked streptavidin chromatographic materials and methods of use thereof
Disclosed herein are chromatographic materials comprising crosslinked streptavidin molecules. The crosslinked streptavidin chromatographic materials may be used to prepare affinity chromatographic materials using biotinylated affinity agents, such as a biotinylated antibody, biotinylated antigen-binding fragment, or biotinylated oligonucleotide. Due to the crosslinked nature of the streptavidin molecules, the materials provided herein have reduced to no streptavidin leachate, resulting in improved performance.
Owner:WATERS TECHNOLOGY CORP
Microorganism bioluminescence detection method and kit
PendingCN122038538AMicrobiological testing/measurementChemiluminescene/bioluminescenceA-DNAStreptavidin
The invention belongs to the technical field of microbial nucleic acid detection and bioluminescence analysis, and discloses a bioluminescence nucleic acid detection method and kit based on DNA-RNA hybrid specific recognition. The kit comprises a streptavidin coated solid phase carrier, a biotinylated DNA capture probe, a signal enhancement DNA, a luminescence report fusion protein and a luminescence substrate thereof, the luminescent reporter fusion protein comprises a DNA-RNA hybrid binding domain and a luminescent enzyme domain. According to the method, target RNA is captured through a solid phase, a DNA-RNA hybrid is formed, the luminous report fusion protein is specifically combined with the DNA-RNA hybrid, a bioluminescence signal is generated after a luminous substrate is added, and the target RNA is detected. In the embodiment, an amplification step is not adopted, so that the method has the advantages of simplicity and convenience in operation, rapidness in detection, lower background and better specificity, and can be used for rapid detection of microorganisms.
Owner:SICHUAN UNIV
Method for efficiently preparing fixed-point biotinylation recombinant protein in escherichia coli body
The invention discloses a method for efficiently preparing fixed-point biotinylation recombinant protein in escherichia coli, and belongs to the technical field of biology. According to the invention, a weakened expression regulatory sequence regulates and encodes a biotin ligase (BirA) gene expression cassette and a target protein gene expression cassette encoding a sequence carrying a biotin receptor peptide tag (AVI-Tag) to be jointly constructed on an escherichia coli expression vector, so that efficient fixed-point biotinylation of a target protein in an escherichia coli body is realized. The method has the advantages of high biotinylation efficiency, high specificity, simplicity and convenience in operation and the like, can be widely applied to the fields of separation and purification of proteins, protein interaction research and the like, and has important practical application value.
Owner:DALIAN UNIV
Ribozyme for labeling biotin on target RNA and screening method and application thereof
The invention relates to ribozyme for labeling biotin on target RNA (Ribonucleic Acid) as well as a screening method and application thereof. The RNA labeling technology is one of core tools for modern molecular biology research and clinical diagnosis. A specific marker is covalently linked to a target RNA molecule, so that accurate tracking, efficient detection and functional analysis of RNA can be realized. A biotin-streptavidin system becomes one of the technical routes which are most widely applied due to extremely high affinity and signal amplification capability of the biotin-streptavidin system. At present, common biotin-labeled RNA technical methods mainly comprise a chemical solid-phase synthesis method, an enzymatic method and a chemical labeling method. However, the methods have the limitations of poor site specificity, low efficiency, complex manipulation and the like in the use process. The engineering ribozyme for catalyzing RNA biotinylation is obtained by taking 12: 0 biotin coenzyme A as a biotin donor through an in-vitro screening technology. The ribozyme provided by the invention can efficiently and specifically carry out biotin labeling on target RNA, the reaction process is simple, and the labeling activity is up to 90% or above.
Owner:ZHEJIANG UNIV
Engineered bispecific exosome preparation with tumor targeted breakthrough capability as well as preparation method and application of engineered bispecific exosome preparation
The invention discloses an engineered exosome preparation with tumor targeting and specific gripper response release functions as well as a preparation method and application of the engineered exosome preparation. The preparation comprises a myocardial cell exosome, an NK cell biotinylation activation antibody, a tumor disease biotinylation antibody, streptavidin, a tumor fiber matrix FAP targeting peptide and a tumor matrix enzyme response peptide, and anchoring is carried out through an acetamide compound-polyethylene glycol anchor point platform. The engineering bispecific exosome preparation with good tumor targeting and barrier breakthrough ability and immune cell activation effect is prepared, and the preparation breaks a solid tumor compact fiber barrier and increases NK cell infiltration by utilizing the fibrosis treatment ability of the myocardial cell exosome and the capture and targeting effects of an anchor point platform; the natural killer cells are in close contact with tumor cells, the lasting tumor cell killing effect is achieved, and the tumor matrix environment is fundamentally changed.
Owner:CHINA PHARM UNIV
Multiple lateral chromatography biological detection system and method based on oligonucleotide functionalized gold nanoparticles
According to the oligonucleotide functionalized gold nanoparticle-based multiple lateral chromatography biological detection system and method provided by the invention, high-sensitivity and programmable nucleic acid target detection is realized through functionalized combination of gold nanoparticles and multiple oligonucleotide probes. The system of the invention comprises: 1) a tunable oligonucleotide-AuNP conjugate (each AuNP is linked to 2-3 thiolated probes); 2) a standardized test strip (containing a streptavidin detection line and an ACTB-828 quality inspection line); and 3) a sandwich hybridization mechanism (the target nucleic acid is combined with the AuNP probe and the biotinylation report oligonucleotide at the same time). The method has the beneficial effects that (1) due to the target-induced plasma aggregation effect, an optical signal is enhanced through AuNP close-range aggregation; and (2) a general test paper structure can adapt to a new target only by adjusting a probe sequence. Experiments show that the detection limit reaches 50 pM, and the room temperature stability is gt; the method is suitable for the fields of rapid pathogen detection, on-site diagnosis and the like.
Owner:INDICATE BIOTECHNOLOGY (SHANGHAI) CO LTD
Method for the generation of an FCRN expressing cell by targeted integration of multiple expression cassettes in a defined organization
ActiveUS12466872B2Solid sorbent liquid separationDepsipeptidesDNA - Deoxyribonucleic acidCarboxysome
Herein is reported a method for producing C-terminally biotinylated FcRn comprising the steps of cultivating a mammalian cell comprising a deoxyribonucleic acid encoding FcRn and E. coli biotin-[acetyl-CoA-carboxylase] ligase (BirA) in a biotin containing medium, and recovering C-terminally biotinylated FcRn from the cell or the cultivation medium, wherein the deoxyribonucleic acid encoding FcRn and E. coli BirA is stably integrated into the genome of the mammalian cell and comprises in 5′- to 3′-direction a first expression cassette encoding class I major histocompatibility complex-like protein (α-FcRn) comprising a HisAvi-tag at the C-terminus, a second expression cassette encoding β2-microglobulin (β2m), a third expression cassette encoding class I major histocompatibility complex-like protein (α-FcRn) comprising a HisAvi-tag at the C-terminus, a fourth expression cassette encoding β2-microglobulin (β2m), and a fifth expression cassette encoding E. coli biotin-[acetyl-CoA-carboxylase] ligase.
Owner:F HOFFMANN LA ROCHE INC
Rapid detection method for cadmium residual quantity in aquatic product based on magnetic qualitative separation and polarized light distinguishing and quantifying
PendingCN121856539AStrong specificityhigh sensitivityPolarisation-affecting propertiesPreparing sample for investigationMicrosphereBiology
The invention relates to a rapid detection method for cadmium residual quantity in aquatic products based on magnetic qualitative separation and polarized light distinguishing and quantifying. The method comprises the following steps: coupling a biotinylated cadmium monoclonal antibody through streptavidin magnetic microspheres, and specifically enriching trace cadmium in the aquatic product digestive juice to form a magnetic compound; a muller matrix microscope is used for collecting a polarization microscopic image of a compound, and a deep learning model (such as RESNET34) is combined to carry out binary classification and particle counting on the image, so that rapid and high-sensitivity quantitative detection of cadmium can be realized. The method is easy and convenient to operate, the target object magnetic separation time is within 2 min, the particle distinguishing accuracy rate is higher than 84%, the quantitative correlation Rreaches 0.9 or above, and the method is suitable for rapid monitoring of trace cadmium (several micrograms / L) in aquatic products.
Owner:FRESHWATER FISHERIES RES CENT OF CHINESE ACAD OF FISHERY SCI
Detection method and kit for detecting anti-Nephrin antibody
The invention relates to the technical field of protein detection, in particular to a detection method and a kit for detecting an anti-Nephrin antibody. The detection method comprises the following steps: capturing an anti-Nephrin antibody in a sample to be detected on an SA matrix by adopting biotinylated Nephrin protein to obtain a first compound; carrying out acidolysis desorption and neutralization on the first compound, and directionally fixing an antibody by Protein G to obtain a second compound; aiming at the second compound, detecting by adopting a biotin-streptavidin-HRP (Horse Radical Polymorphism) system; the acidolysis desorption comprises the following steps: treating the first compound for 10-20 minutes by adopting an acid solution with the pH value of 2-3 under the conditions that the temperature is 35-40 DEG C and the speed is 500-800 rpm. The method provided by the invention can accurately and quickly detect the level of the anti-Nephrin antibody in the sample, and has important application value.
Owner:THE FIRST MEDICAL CENT CHINESE PLA GENERAL HOSPITAL
Chemiluminiscence detection kit for urine and bladder cancer antigen and preparation method of chemiluminiscence detection kit
The invention provides a UBC-CK8 / 18 detection kit based on chemiluminescence, and the kit is characterized in that an antibody is immobilized through a streptavidin-biotin system to form a magnetic bead-biotinylated antibody-antigen-acridinium ester labeled antibody compound; after washing, the substrate solution excites the acridinium ester to generate a photon signal, and a relative luminescence unit (RLUs) is captured by a full-automatic chemiluminescence immunoassay instrument, so that rapid and low-cost clinical detection of bladder cancer is realized, and the detection performance is good.
Owner:AFFILIATED HOSPITAL OF JIANGNAN UNIV
Streptavidin chromatographic materials and methods of use thereof
Disclosed herein are chromatographic materials comprising streptavidin molecules, wherein the streptavidin molecules are bound to biotin and one or more biotinylated affinity agents. The biotinylated affinity agent may be a biotinylated antibody, biotinylated antigen-binding fragment, or biotinylated oligonucleotide. The streptavidin chromatographic materials may be used to prepare affinity chromatographic materials. Due to biotin binding to the conjugated streptavidin, the materials provided herein have reduced to no streptavidin leachate, resulting in improved performance.
Owner:WATERS TECHNOLOGY CORP
Cell Selection Chamber And Methods Of Cell Selection
PendingUS20260053997A1Other blood circulation devicesMedical devicesCell selectionMechanical engineering
A cell selection chamber includes an inlet and an outlet, with a reservoir positioned therebetween and configured to allow for fluid flow from the inlet to the outlet. The reservoir contains a plurality of microbubbles, which are each at least partially coated with streptavidin. An upstream membrane is positioned between the inlet and the reservoir, while a downstream membrane is positioned between the outlet and the reservoir. The membranes include a plurality of pores each having a diameter that is less than a diameter of the microbubbles. In use, a heterogeneous population of cells is mixed with a biotinylated antibody additive and then conveyed into the cell selection chamber so as to cause target cells of the heterogeneous population of cells to become bound to the microbubbles and to cause other cells of the heterogeneous population of cells to flow out of the cell selection chamber as unbound cells.
Owner:FENWAL INC
Neuron labeling method and application thereof in neuron single cell reconstruction
PendingCN121022753APolypeptide with localisation/targeting motifAntibody mimetics/scaffoldsStainingMarker enzymes
The invention relates to a method for marking neurons and application of the neurons to unicellular reconstruction of the neurons. The method comprises the following steps: expressing a proximity marker enzyme with enhanced solubility in a target neuron, and providing biotin to the target neuron so as to form biotinylated protein in the neuron; carrying out integral sample dyeing on the biotinylated protein by using monovalent streptavidin coupled with a detectable marker; imaging the dyed sample to obtain a neuron structure image; and reconstructing the morphological structure of the single neuron based on the image data.
Owner:NAT INST OF BIOLOGICAL SCI BEIJING
Mutant gene enrichment method, detection method and kit based on Cas9 specific cleavage and isonucleotide magnetic beads
The invention discloses a mutant gene enrichment method, a mutant gene detection method and a mutant gene detection kit based on Cas9 specific cleavage and isonucleotide magnetic beads. The method comprises the following steps: firstly, extracting free DNA; then identifying and cutting a completely matched wild type sequence by using Cas9 protein and specific sgRNA, and reserving the mutant DNA due to base mismatch; cas9 treated DNA and streptavidin magnetic beads containing a biotinylated oligonucleotide probe are incubated, guanine and cytosine in the probe are substituted by isoguanine and isocytosine respectively, and adenine and thymine are modified by locked nucleic acid, so that high-stability homodromous pairing is realized, and mutation DNA is selectively enriched; and finally, carrying out qPCR or sequencing detection on the enriched DNA. The kit has the characteristics of simplicity and convenience in operation, high specificity and high sensitivity, can be used for detecting single-base-level mutation, and is suitable for liquid biopsy, tumor early screening and genetic disease mutation detection.
Owner:SUZHOU HAIMIAO BIOTECH CO LTD
Protein detection method and kit for implementing same
The invention provides a protein detection method and a kit for implementing the method, and the protein detection method comprises the following steps: adding antibody-coated capture beads into a sample, and capturing target protein molecules; a biotinylated detection antibody and streptavidin-multi-HRP (horse radish peroxidase) are used for marking captured target protein molecules to form a complete enzyme-labeled immune complex; in a solution containing hydrogen peroxide and a tyrosine-fluorophore conjugate, generating a covalently linked fluorescent dye; fixing the beads in the fibrous protein hydrogel to form a fibrous protein hydrogel layer for bead immobilization; capturing bright field and fluorescence images of the bead array, and analyzing the images for single molecule counting. According to the method, the casein is used for signal amplification, the step of signal amplification is achieved on the beads, and the fluorophore is directly combined with the beads, so that the requirement for separating the beads in a micro-pore array or micro-fluid liquid drops is eliminated, and the requirements for expensive equipment and complex micro-fluid or robot technology are reduced.
Owner:SUZHOU YUANXING INTELLIGENT MANUFACTURING TECHNOLOGY CO LTD
Single-molecule fluorescence immunoassay kit for detecting GFAP protein of dry plasma spot and preparation method of single-molecule fluorescence immunoassay kit
PendingCN120761624AColor/spectral properties measurementsBiological testingProtein detectionMagnetic bead
The invention discloses a single-molecule fluorescence immunoassay kit for detecting dry plasma spot GFAP protein and a preparation method, and belongs to the technical field of immunodetection. The kit provided by the invention comprises an anti-GFAP mouse monoclonal antibody coated magnetic bead reagent, a biotinylated anti-GFAP mouse monoclonal antibody reagent, a streptavidin-beta-galactosidase reagent, a resorufin beta-D-galactoside reagent, a dry plasma spot eluent, a GFAP sample diluent, a GFAP calibration product, a GFAP quality control product and dry plasma spot filter paper. The dry plasma spot eluent contains PEG (Polyethylene Glycol) 6000 with the concentration of 15g / L to 25g / L, NaCl with the concentration of 15g / L to 20g / L, HEPES with the concentration of 11g / L to 12g / L, Triton X-100 with the concentration of 10g / L to 12g / L, EDTA (Ethylene Diamine Tetraacetic Acid) with the concentration of 4.5 mmol / L to 5.5 mmol / L, EGTA (Ethylene Glycol Terephthalate) with the concentration of 1.5 g / L to 2.0 g / L and BSA (Bovine Serum Albumin) with the concentration of 1.2 g / L to 1.8 g / L, and the pH value is adjusted to 7.3 to 7.5 by NaOH; aiming at the bottleneck problem of trace GFAP protein detection in a dried plasma spot sample, the detection sensitivity, accuracy and filter paper applicability are improved by optimizing an eluent formula, diluent components and a detection system.
Owner:YOUDA BIOTECHNOLOGY (SHANGHAI) CO LTD
Detection method of flow fluorescent single molecule pTau217
The invention discloses a flow fluorescent single molecule pTau217 detection method, which comprises the following steps: reacting aminated polystyrene microspheres with diphenyl cyclooctyne-disulfide bond-active ester and azide functionalized streptavidin in sequence to prepare a composite microsphere dispersion liquid; the method comprises the following steps: reacting an elisa plate with 1-ethyl-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide, and coupling and capturing an antibody; sequentially adding a protein sample and a biotinylation detection antibody into the enzyme plate coupled with the capture antibody for reaction, and cleaning; adding composite microsphere dispersion liquid, oscillating and cleaning; adding a disulfide bond reducing agent for room-temperature reaction; and collecting the reacted solution, and counting the microspheres in the solution to realize the detection of the content of the flow fluorescent single molecule pTau217. According to the detection method disclosed by the invention, the detection on the quantity of the pTau217 protein can be realized, and the detection sensitivity is relatively high; the related operation method is simple, the detection cost is low, and the method is suitable for popularization and application.
Owner:WUHAN WISE DIAGNOSTIC TECH CO LTD
Mycobacterium tuberculosis detection method and kit
PendingCN121496079AMicrobiological testing/measurementMicroorganism based processesMagnetic beadStreptavidin
The invention belongs to the technical field of biology, and provides a mycobacterium tuberculosis detection method and a kit, the detection method comprises the following steps: adding a to-be-detected sample containing mycobacterium tuberculosis into a lysis working solution, carrying out a heat preservation reaction, centrifuging, and taking a supernatant, the cracking working solution contains a biotinylated metal complexing agent and biotinylated nanoparticles, and guanidine isothiocyanate and hydrophobic alkyl groups are connected to the biotinylated nanoparticles; adding a magnetic bead suspension into the supernatant, stirring at room temperature for reaction, standing, taking an upper solution, adding the upper solution into a purification column, purifying and washing to obtain a detection sample, and the surface of a magnetic bead contains streptavidin; and adding a detection sample into an amplification reagent for fluorescent PCR amplification, and analyzing a PCR amplification product by using a fluorescence melting curve to finish detection. The method can effectively reduce impurities contained in the detection sample and improve the detection accuracy.
Owner:ZHUHAI ENCODE MEDICAL ENG
Polar aprotic organic solvent for dissociating streptavidin-biotin conjugate as well as screening method and application of polar aprotic organic solvent
The invention relates to a polar aprotic organic solvent for dissociating a streptavidin-biotin conjugate as well as a screening method and application of the polar aprotic organic solvent. Aiming at the problems that a streptavidin-biotin conjugate system is difficult to elute quickly under a mild condition and a carrier is difficult to regenerate, a solvent molecule descriptor database is constructed; a polar aprotic organic solvent which meets specific molar refractive index and polarity parameters and is a non-hydrogen bond donor is screened from a large number of candidate solvents by utilizing a machine learning model. The solvent can realize rapid reversible dissociation of biotin conjugates from streptavidin at room temperature, and the carrier still keeps high activity after multiple rounds of binding-elution cycles. The solvent is further used for purification of biotinylated small molecules in a crude reaction solution, enrichment and elution of mitochondria and cell surface samples and the like, topological resolution proteomics analysis is achieved, a high-purity product can be obtained in a high-flux mode, and site level topological analysis can be completed.
Owner:SHANGHAI JIAOTONG UNIV
PRRSV (porcine reproductive and respiratory syndrome virus) N protein monoclonal antibody as well as preparation method and application thereof
PendingCN121698996ASsRNA viruses positive-senseAntibody mimetics/scaffoldsPorcine reproductive and respiratory syndrome virusBiotin
The invention discloses a PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) N protein monoclonal antibody as well as a preparation method and application thereof, belongs to the technical field of biological materials, and solves the problem that in the prior art, the stability and specificity of a PRRSV monoclonal antibody screening result are not ideal enough. The method comprises the following steps: constructing a gene of a PRRSV N protein into a plasmid, and performing expression and purification to obtain a recombinant N protein; uniformly mixing the recombinant N protein, a Modifer reagent, a Quencher reagent and a DR-B reagent, centrifuging, taking supernate, re-suspending the supernate in an SB reagent, and separating out biotinylated N protein through a membrane; and screening the positive B cells combined with the biotinylated N protein by using a Hypercell high-throughput single cell screening platform to obtain the monoclonal antibody. The monoclonal antibody aiming at the PRRSV N protein is prepared by a single B cell screening technology, and a foundation is laid for PRRSV diagnosis and related research of N protein functions.
Owner:JINLIN MEDICAL COLLEGE
Reagents for detecting varicella-zoster virus infection and methods of making same
ActiveCN116539875BVirus peptidesNucleic acid vectorVaricella-zoster virus infectionSerum protein albumin
The present application relates to the field of immunological detection, and particularly relates to a reagent for detecting varicella-zoster virus infection and a preparation method thereof. The reagent comprises R1 reagent and R2 reagent; the R1 reagent is a working solution of streptavidin magnetic beads, and the preparation method comprises the following steps: firstly coupling activated carboxyl magnetic beads with streptavidin; dialyzing the magnetic beads obtained by the first coupling to remove streptavidin physically combined with the magnetic beads; secondly coupling the dialyzed magnetic beads with streptavidin; quenching the magnetic beads obtained by the second coupling, and then placing the magnetic beads in a working solution to obtain the working solution; the working solution contains fetal bovine serum, bovine serum albumin and lysine; the R2 reagent is a biotinylated VZV gE recombinant protein, and the VZV gE recombinant protein is a varicella-zoster virus gE glycoprotein extracellular domain fragment expressed by human embryonic kidney cells. The reagent has high sensitivity and specificity in the detection of varicella-zoster virus infection.
Owner:PEKING UNIV
A method and a kit for generating nucleic acid for target capture
The invention relates to a method and a kit for generating nucleic acid for target capture. The method involves binding of a circ-olio sequence to at least one target oligonucleotide to generate a single stranded circular DNA. The single stranded circular DNA is bidirectionally amplified with the sense and antisense oligonucleotides, followed by biotinylation to obtain for a double-stranded concatemer of linear sequence. The double-stranded concatemer of linear sequence with the restriction endonucleases produces a target capture nucleic acid. The method offers a scalable targeted capture of pr eselected genomic regions, and is cost effective. The method of generating nucleic acid for target capture has application in the field of oncology hematology, inherited malignancies, germline disorders, microbiology and prenatal diagnosis. The invention generates baits over high GC-rich regions of the genome.
Owner:TATA MEMORIAL CENTRE-ADVANCED CENTRE FOR TREATMENT RESEARCH & EDUCATION IN CANCER (TMC-ACTREC)
Anti-CD4 antibody and microsphere flow-type immunofluorescence luminescence detection kit and application thereof
PendingCN121699012AImmunoglobulins against cell receptors/antigens/surface-determinantsBiological testingAutoimmune conditionBinding site
The invention provides an anti-CD4 antibody as well as a microsphere flow immunofluorescence luminescence detection kit and application thereof, and provides two monoclonal antibodies for recognizing different binding sites of a CD4 antigen, an antibody coupling system based on magnetic fluorescent microspheres, a biotinylation detection antibody and phycoerythrin labeled streptavidin. The invention also discloses a corresponding detection method and a preparation method of the kit. The CD4 antigen detection kit and the antibody combination thereof provided by the invention can significantly improve the sensitivity, stability, specificity and flux capacity of CD4 detection, and are suitable for multiple fields of immune function evaluation, autoimmune disease evaluation, inflammatory disease research, HIV infection auxiliary diagnosis and the like.
Owner:THE STOMATOLOGIAL HOSPITAL OF ZHEJIANG UNIV SCHOOL OF MEDICINE