66 results about "Nonspecific binding" patented technology

The invention belongs to the field of molecular sieve material characterization, and particularly relates to a method for analyzing aluminum atom pairs in a ZSM-5 molecular sieve based on super-resolution fluorescence microscopic imaging, which comprises the following steps: activating a ZSM-5 molecular sieve sample; contacting the obtained sample with probe molecules; washing the ZSM-5 molecular sieve sample, and removing unbound or nonspecifically bound probe molecules; imaging the marked sample by using a super-resolution fluorescence microscope to obtain an image sequence containing a single fluorescent molecule emission event; processing the image sequence, determining a space coordinate of each fluorescence event, and reconstructing a super-resolution fluorescence image exceeding an optical diffraction limit; and based on the space coordinates of the fluorescent dots in the super-resolution fluorescence image, carrying out statistics to obtain the position, surface density or distribution information of the aluminum atom pairs in the ZSM-5 molecular sieve. According to the method disclosed by the invention, nanoscale spatial resolution, direct imaging and quantitative analysis of Al-pair in ZSM-5 (Zeolite Socony Mobil-5) can be realized.

The invention provides a nucleic acid probe, a primer group, a kit, a detection method and application, and relates to the technical field of biology. The nucleotide sequence of the nucleic acid probe is as shown in SEQ ID NO. 1; wherein the 3'end of the nucleic acid probe is modified with MGB, and locked nucleic acid is modified at a base position corresponding to the difference between the swine fever wild strain and the swine fever vaccine strain in the nucleotide sequence. According to the nucleic acid probe, a specific sequence is utilized to precisely target a swine fever wild strain, and locked nucleic acid modification is introduced at a differential site, so that the recognition capability on single-base mismatch is remarkably enhanced, non-specific binding to a vaccine strain is thoroughly blocked, and false positive interference is eliminated; and 3 '-terminal MGB modification is matched to improve the binding stability and sensitivity, so that the efficient and accurate identification of the swine fever wild strain under the immune background is ensured.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody combination for detecting porcine epidemic diarrhea virus NP protein and application of the monoclonal antibody combination. The combination is composed of monoclonal antibodies 1E11 and 5C8 and can efficiently and specifically recognize porcine epidemic diarrhea virus NP protein, amino acid sequences of complementary determining regions of heavy chain and light chain variable regions of the combination are clear and are shown as SEQ ID NO.1-12 respectively, high specificity and high affinity of the antibodies are guaranteed, the antibody combination can effectively capture and detect target antigens, and the antibody combination can be used for detecting the porcine epidemic diarrhea virus NP protein. The risk of non-specific binding and cross reaction is reduced. The combination shows excellent sensitivity and specificity in detection platforms such as colloidal gold immunochromatography and the like, and provides a stable and reliable biological recognition tool for rapid and accurate detection of the porcine epidemic diarrhea virus.

The invention belongs to the technical field of immunohistochemical detection, and discloses a modified composite sealing buffer solution as well as a preparation method and detection application thereof. The modified composite blocking buffer solution comprises a basic buffer system, a composite blocking agent, a nonionic detergent, an endogenous interferent blocking agent and an antibody stabilizer, wherein the basic buffer system provides a stable pH environment and ionic strength; the composite sealing agent firstly occupies a shielding site, and the background is lowered; the nonionic detergent destroys hydrophobic interaction, effectively elute and loosen adsorbed molecules, and promote dissolution of components with relatively strong hydrophobicity; the endogenous interferent blocker can be combined with a receptor firstly, so that non-specific combination of an antibody and the receptor is blocked, and the problem of specific background is solved; the antibody stabilizer can protect the activity of the detection reagent and maintain the stability, and the obtained modified composite sealing buffer solution and the kit thereof have the advantages of multiple cooperative sealing mechanisms, prospective blocking of endogenous interference, improvement of the stability, compatibility and high efficiency of the antibody and the like.

The present invention relates to a polymer conjugate comprising a ULA polymer a) and one or more polypeptides b), wherein the polypeptides are conjugated to the ULA polymer, and the conjugate is coated on the surface of a device for culturing cells. The peptides can interact with receptors on the surface of the cells, while the polymer prevents non-specific binding of the cells to the surface of the device.

The invention discloses a detection method of inflammatory marker protein. According to the method, the micro-fluidic chip with a sample loading area, a buffer treatment area, a sample channel and a detection area is prepared on filter paper through wax printing; chitosan is preset in the sample loading area and is used for treating a sample, and bromothymol blue is fixed in the detection area and is used as a pH indicator. During detection, a sample is driven by a phosphate buffer solution with the pH value of 6.0 to flow to a detection area through capillary action, the C-reactive protein and the indicator are subjected to non-specific binding, so that the color of the detection area is changed from yellow to green, the length of a green developing signal is in direct proportion to the concentration of the CRP, and quantitative detection can be realized by measuring the length. The method does not need an antibody, is low in cost, is simple and convenient to operate, can complete detection within 15 minutes, has the detection limit of 1.56 ng / ml and the linear range of 5-25 ng / ml, and is suitable for rapid screening and field detection of basic medical institutions.

The invention belongs to the technical field of electrochemical analysis and detection, and particularly relates to an electrochemical immunosensor for detecting adiponectin as well as a preparation method and application of the electrochemical immunosensor. The preparation method comprises the following steps: (1) connecting a screen-printed electrode SPCE to an electrochemical workstation by using a 0.01 M PBS (Phosphate Buffer Solution) with the pH value of 7.4, performing-0.2 V-1. 2V cyclic voltammetry scanning, and activating for 10 circles under 50mV / S; and (2) by taking 0.01-1M H2SO4 as a base solution, connecting the screen-printed electrode SPCE to an electrochemical workstation, carrying out electro-polymerization on poly-o-phenylenediamine in a 50-100mM o-phenylenediamine and NH2-UiO-66 solution to form a poly-o-phenylenediamine and NH2-UiO-66 composite modified electrode, washing with deionized water, and drying at room temperature. (3) soaking the modified electrode in a glutaraldehyde solution for 10-70 minutes, cleaning with deionized water, drying at room temperature, adding 10-100 [mu] L of ADPN antibody solution and 100-600 [mu] g / mL of ADPN antibody solution, and incubating; and (4) washing with ionized water, drying at room temperature, preventing non-specific binding sites at 37 DEG C for 0.5-1.5 hours by using 3-10 microliters of 0.8-1.2 wt% casein solution, and washing the screen-printed electrode again to obtain the electrochemical immunosensor.

A high-throughput screening system and method for small molecule-protein interactions includes first immobilizing selected small molecules on a solid matrix. Immobilization is preferably performed via photoaffinity labeling, where a linker is used to bind the small molecule to the matrix. cDNA libraries are constructed based on various human tissues. Tissue mRNA is transcribed into cDNA and transferred to a phage genome to construct a phage display library, which is then mixed with small molecules conjugated to microspheres and incubated. The incubated mixture is washed to remove unbound phages, including non-specific and weakly bound phages. Bound phages are amplified and used for the next round of biopanning. The biopanning cycle is repeated, and then PCR is performed on each plaque obtained from the final round of biopanning. The PCR products from each small molecule experiment are barcoded, merged, and then subjected to next-generation sequencing.

The invention belongs to the technical field of biotechnology and protein determination, and particularly relates to confining liquid as well as a preparation method and application thereof. Modified bovine serum albumin and recombinant casein polypeptide are used as core components of the confining liquid, hydroxypropyl-beta-cyclodextrin, polyglycerol fatty acid ester and other auxiliary components are added, and the confining liquid is prepared through a specific technology. The modified bovine serum albumin and the recombinant casein polypeptide have a synergistic effect, so that active sites of the detection carrier can be efficiently sealed; the stabilizing agent and other components are supplemented, so that the stability and compatibility of the confining liquid are improved. The confining liquid is simple and convenient in preparation process, controllable in cost, suitable for various detection carriers such as elisa plates and nitrocellulose membranes, capable of reducing non-specific binding and improving detection sensitivity, and suitable for various biological detection scenes such as enzyme-linked immunosorbent assay.

The present invention relates to an antibody fragment consisting of only the constant region of an antibody, and more particularly to an antibody fragment having no variable region of an antibody and consisting of only a heavy chain constant region (C H ) and a light chain constant region (C L ). For example, the present invention relates to an antibody fragment consisting of a heavy chain constant region and a light chain constant region (IgCw-gamma1epsilon2-4 / kappa), in which a gamma constant region (Cgamma1) and an epsilon constant region (Cepsilon2-4) are fused; a nucleic acid encoding the antibody fragment; a kit comprising the antibody fragment; and a use of the antibody fragment. In order to evaluate the efficacy of an antibody in the development of a new antibody drug or the like, a control, i.e., a reference antibody, is required, and the existing reference antibody cannot be completely free from non-specific binding, i.e., off-target effects due to reactions with other antigens, and the purification yield is also low. On the other hand, when the recombinant protein IgCw-gamma1epsilon2-4 / kappa consisting of only the constant region of an antibody according to the present invention is used, the problems of the existing reference antibody can be overcome, and the recombinant protein can also be used as an Fc epsilon receptor inhibitor that inhibits an allergic reaction. Therefore, the IgCw-gamma1epsilon 2‑4 / kappa according to the present invention can be used in various ways in the field of biomedical science.

The invention discloses a screening method and application of an aptamer highly specifically bound with uPAR, and relates to the technical field of biology.The screening method of the aptamer comprises the steps that S1, uPAR protein with a his tag, a single-chain nucleotide library containing basic groups and PCR primers are prepared; and S2, dissolving the uPAR protein in a coupling buffer solution, combining the uPAR protein with the Ni-band overnight, filtering, and calculating the amount of the protein coupled to the Ni-band. According to the screening method provided by the invention, through multiple rounds of screening, a non-specific binding sequence is removed by using Ni-bead, so that the binding specificity and affinity of the screened aptamer and the uPAR protein are remarkably improved, and the aptamer with high specificity and high affinity has important application value in disease diagnosis, targeted therapy and drug research and development, and has a wide application prospect. Compared with the traditional detection method based on antigen-antibody reaction, the screening method provided by the invention has detailed steps and can be optimized and adjusted according to needs to adapt to different experimental requirements and targets as the aptamer is used as a single-stranded DNA or RNA molecule.

The invention discloses an optical fiber sensor for detecting the concentration of exhaled gas anesthetic and a detection system thereof, and relates to the technical field of anesthetic detection and optical fiber sensors, and the optical fiber sensor comprises an input optical fiber, a U-shaped tapered optical fiber and an output optical fiber which are connected in sequence; a specific recognition layer for detecting the anesthetic is arranged outside the U-shaped tapered optical fiber, and a functional treatment layer for closing a non-specific binding point of the anesthetic is arranged outside the specific recognition layer; the specific recognition layer is obtained by sequentially modifying the surface of the U-shaped tapered optical fiber with a sodium hydroxide solution, an aminopropyltriethoxysilane solution, a glutaraldehyde solution and an anesthetic specific binding molecular solution; the functionalized treatment layer is obtained by sealing binding sites of anesthetic specific binding molecules which are not bound on the surface of the specific recognition layer by adopting a bovine serum albumin solution. The problems that an existing optical fiber sensor is insufficient in sensitivity, poor in selectivity and difficult to integrate clinically are effectively solved.

The invention discloses a porcine rotavirus VP4 protein monoclonal antibody, a kit and application, and belongs to the field of porcine rotavirus antibody detection. The porcine rotavirus VP4 protein monoclonal antibody comprises a heavy chain and a light chain, the amino acid sequence of the heavy chain is shown as SEQ ID NO. 1, and the amino acid sequence of the light chain is shown as SEQ ID NO. 2. According to the present invention, the blocking ELISA detection kit and the detection method are innovatively constructed by using the monoclonal antibody targeting the VP4 conserved functional epitope as the core, and the method can significantly reduce the cross reaction and truly reflect the protective immune level; compared with the prior art, the method provided by the invention has the advantages of no need of species-specific secondary antibody, avoidance of non-specific binding and serum source interference, strong universality, good repeatability and suitability for high-throughput application, fills the technical blank, and provides a novel tool with high specificity, high correlation and high practicability for precise immune monitoring and vaccine evaluation of PoRV.

The application discloses a latex-enhanced immunoturbidimetric detection reagent and application of a serine protease in the latex-enhanced immunoturbidimetric detection reagent, relates to the technical field of biological detection, and can reduce the interference of a matrix component on the binding of a target and an antibody, reduce the probability of non-specific binding, and improve the accuracy of detection by pretreating a to-be-detected sample by using a reagent R1 containing the serine protease; the difference between a sample matrix and a quality control product matrix can be avoided, the accuracy and reliability of a detection result are improved; the difference between different quality control product matrices can be eliminated, the difference between standard curves obtained by using different matrix quality control products is reduced, and the compatibility of three-party quality control is enhanced.

This invention belongs to the field of biodetection technology, specifically relating to a material for detecting anti-NF155 antibodies and its preparation method. This invention constructs an expression plasmid capable of highly expressing the NF155 antigen, transfects it into HEK293T cells, and modifies the pCDH vector with enhancers during construction. Subsequently, a modified cell culture protocol is used to obtain a material for detecting anti-NF155 antibodies based on a cell immunofluorescence methodology. This invention uses a cell immunofluorescence methodology to maximize the preservation of the antigen's spatial conformation through cell expression and antigen immobilization, preventing false negatives due to changes in binding sites. This invention selects the lentiviral vector pCDH to avoid the problem of non-specific binding that is difficult to distinguish, as the proteins expressed by common target gene vectors such as pcDNA3.1 may themselves contain antibody binding sites. This improves detection specificity and solves the problem of false negatives.

The application provides a kit and method for detecting common fusion genes of lymphoid leukemia based on a multiple digital PCR method. By constructing a synergistic mechanism of primers and fluorescent probes, optimizing the collocation of primers, probes and fluorescent markers, and effectively avoiding the non-specific binding between amplification products. The innovative introduction of a composite reaction aid with a specific formula, namely a PCR enhancer, significantly improves the amplification efficiency and specificity of the multiple detection system. The kit provided by the application realizes the integrated detection capability of seven fluorescent channels, Atto 425, VIC, FAM, ROX, CY5, CY5.5 and CY7 in a single hole design. By innovatively constructing a ROX+CY5 and Atto 425+VIC dual-channel combined detection system, when the ROX and CY5 channels or the Atto 425 and VIC channels in the same detection hole simultaneously present positive signals, two additional detection targets can be specifically interpreted. This innovative detection strategy based on channel combination enables the kit to break through and achieve accurate detection of 18 fusion genes with only 2 detection holes.

The application relates to a preparation method of an electrochemiluminescence sensor and application thereof, and belongs to the technical field of electrochemiluminescence sensors, and specifically comprises the following steps: step 1, firstly, a bare glassy carbon electrode is repeatedly polished with aluminum oxide powder, and then the electrode is thoroughly cleaned before use; Au@CN solution is coated on the electrode; step 2, after the electrode is dried, a conconavalin A solution is added dropwise, and after 1 hour, the electrode is washed with a phosphate solution to block the non-specific binding between the conconavalin A and the electrode surface; step 3, NMS / CNT@Glu solution is coated on the electrode surface, and after 1 hour, the obtained electrode is washed to remove the unbound NMS / CNT@Glu on the outer surface, so that an electrochemiluminescence sensor for detecting conconavalin A is obtained. When the sensor detects conconavalin A, a wide linear range and a low detection limit are exhibited, so that the sensor can be applied to actual detection.

The invention discloses differential temperature comparison single molecule detection based on a reversible thermal change renaturation query probe, and relates to the field of single molecule immunodiagnosis. According to the invention, the property of repeated thermal denaturation / renaturation of the query probe is utilized, and the specific binding activity of the query probe is controlled, so that a specific binding molecular signal and a non-specific binding molecular signal of the query probe are distinguished, and a novel high-specificity single-molecule immunodiagnosis method is provided.

The application provides a hepatitis E virus nucleic acid detection primer, probe and kit, wherein according to a target sequence of a hepatitis E virus gene, a primer and a probe sequence in an asymmetric primer ERA detection system are designed, screened, and optimized in terms of an upstream primer ratio, a downstream primer ratio and an amplification reaction system, so that the primer and the probe can specifically recognize a nucleic acid sequence of the hepatitis E virus, non-specific binding with other irrelevant nucleic acids is reduced, and thus the specificity of detection is improved; meanwhile, efficient amplification of the hepatitis E virus nucleic acid is realized, and even in the case of extremely low content of the virus nucleic acid, the hepatitis E virus nucleic acid can be accurately detected, and thus the sensitivity and the amplification efficiency of detection are improved.

This invention belongs to the field of immunoassay technology, specifically relating to acetylpyridinium hapten, complete antigen, antibody, preparation methods, and applications. The acetylpyridinium hapten provided by this invention has the structure shown in Formula I, where R1 is an active group capable of direct coupling with a carrier protein. The acetylpyridinium hapten provided by this invention retains the basic structure of acetylpyridinium, using the original -CH2-CH2-CH2-CH2- structure of acetylpyridinium as a linker arm. An active group R1 capable of direct coupling with a carrier protein is introduced into the linker arm at a site far from the characteristic structure of acetylpyridinium, effectively avoiding non-specific binding caused by the external linker arm. After coupling with the carrier protein, the introduced active group can fully expose the characteristic structure of the acetylpyridinium bulk molecule. The prepared acetylpyridinium complete antigen is immunogenic, and the prepared anti-acetylpyridinium antibody has good specificity and high sensitivity. It can be used to establish immunological detection methods or reagents for acetylpyridinium, and has broad application prospects.

The present invention relates to a method for quantifying target polynucleotides, particularly RNA molecules such as 16S rRNA, through a process that converts the abundance of target molecules into the abundance of predefined barcoded oligonucleotides The invention enables reduced nonspecific binding, protocol flexibility with pausing and re-entry points, and versatile, optionally multiplexed data acquisition including flow-cytometry-like or imaging-based measurements.

The present invention relates to a very short peptide sequence having pH-selective binding ability to FcRn. The peptide of the present invention is not large in size like an Fc region, thereby enabling easy penetration into target cells and / or tissues (for example, solid tumors). In addition, since N-glycosylation is not essential, the peptide can be produced in inexpensive Escherichia coli rather than animal cells. Furthermore, the peptide does not bind to FcγR and thus does not exhibit off-target toxicity, and has no possibility of forming undesired combinations, thereby maximizing production efficiency.

The invention relates to a double-color fluorescence-based gel electrophoresis detection method, device, medium and kit, and the method comprises the following steps: marking a standard sample with a first fluorescent marker through covalent modification, carrying out mixed pre-dyeing on a test sample and a non-specifically bound second fluorescent dye, and loading the standard sample and the test sample to the same gel lane for synchronous electrophoresis; after electrophoresis, a dual-channel fluorescence imaging system is used for synchronously acquiring exclusive images corresponding to the two fluorescent substances; based on the migration position of the standard sample, calculating a fluorescence signal intensity proportion to eliminate crosstalk, establishing a fragment length and migration position fitting model, and finally accurately calculating the length of a target nucleic acid fragment in the test sample; according to the method, errors between lanes are eliminated through synchronous detection of the same lane, the operation process is simplified, the detection accuracy and efficiency are remarkably improved, and the method is suitable for separation and analysis of biological macromolecules such as DNA and RNA and can be widely applied to the fields of molecular biology research, clinical diagnosis and the like.

Methods and formulations for preparing low non-specific binding surfaces are described, and the prepared surface can provide improved performance for nucleic acid detection and base calling applications. The surface provides more accurate nucleic acid detection, enhanced contrast to noise ratio, and better data collection.

The invention relates to methods for conducting solid-phase binding assays. One example is an assay method having improved analyte specificity where specificity is limited by the presence of non-specific binding interactions.

The invention discloses a hybridization chain reaction fluorescent biosensor and a preparation method and application thereof, red cell membrane coated gold nanoparticles are used as a reaction substrate, the surface of the gold nanoparticles is covalently connected with a sulfydryl modification initiation probe aiming at target miRNA through an Au-S bond, and the red cell membrane coated gold nanoparticles are treated by Fe < 3 + >. According to the method, the red cell membrane completely coats the gold nanoparticles, so that common false positive signals in a complex body fluid environment are remarkably reduced; the hybridization chain reaction is limited on the surface of the cell membrane, so that the local concentration of the hairpin probe is improved, the chain amplification process is accelerated, and the detection time is shortened; sufficient sites are provided for probe fixation and HCR polymerization by utilizing the large specific surface area of the gold nanoparticles, and the signal amplification factor is further enhanced; fe < 3 + > is introduced to perform surface modification on the red cell membrane coated gold nanoparticles, so that non-specific binding is effectively inhibited, blank background fluorescence is remarkably reduced, and the signal-to-noise ratio is increased.

The invention relates to methods for conducting solid-phase binding assays. One example is an assay method having improved analyte specificity where specificity is limited by the presence of non-specific binding interactions.

The invention provides a sample pad treating fluid, immunochromatography test paper, a kit and application thereof in detection of cytomegalovirus, and relates to the technical field of biological detection.The sample pad treating fluid comprises 0.05-0.2 wt% of sodium carbonate, 3.0-5.0 wt% of borax, 0.5-2.0 wt% of PVP, 0.5-2.0 wt% of T-casein and more than or equal to 0.5 mg / mL of blocking agent, and after the blocking agent is added, the detection specificity is improved. The technical problem of low specificity caused by non-specific binding in detection of cytomegalovirus in a breast milk sample in the prior art is solved. The method can be used for preparing products for detecting cytomegalovirus. The detection process is rapid (about 5-15 minutes), no instrument is needed, and the method is suitable for field detection. The sensitivity of the immunochromatography test paper is improved and can reach 20 mu g / ml. The rapid detection of the CMV antigen in the breast milk is realized.

This invention discloses an anti-ERBB2 monoclonal antibody, its preparation method, and its applications, belonging to the field of biomedical technology. This anti-ERBB2 monoclonal antibody uses the intracellular domain of the human ERBB2 protein as an immunogen and is obtained through animal immunization, B cell screening, gene cloning, and recombinant expression techniques. It can specifically recognize the human ERBB2 protein, exhibiting high affinity and high specificity, and can recognize both native and denatured ERBB2 proteins. This monoclonal antibody can be applied in immunohistochemistry, immunofluorescence, Western blotting (WB), flow cytometry, ELISA detection, and tumor targeted therapy. Experimental results show that this antibody has low background and no non-specific binding, accurately detecting ERBB2-overexpressing tumor cells and tissues, and has significant application value in the diagnosis, prognostic assessment, and targeted therapy of ERBB2-related diseases such as breast cancer.

The invention discloses a portable electrochemical immunosensor and a preparation method and application thereof.The portable electrochemical immunosensor comprises a plurality of working electrodes, counter electrodes and reference electrodes which are integrally integrated on a PET substrate, a carbon-based material is deposited on the surface of one of the working electrodes, the carbon-based material is covalently connected with an influenza A H1N1 virus antibody through a cross-linking agent, and the counter electrodes and the reference electrodes are arranged on the surface of the other working electrode. A non-specific binding site of the influenza A H1N1 virus antibody on the surface of the working electrode is sealed by bovine serum albumin. According to the invention, the SPCE based on the PET substrate integrates a working electrode, a counter electrode and a reference electrode, and is precisely packaged through insulating ink. The integrated design not only greatly reduces the production cost, but also enables the sensor to have excellent mass production performance, so that the sensor is especially suitable for developing disposable and portable rapid detection devices.