56results about How to "High coupling rate" patented technology

The invention relates to the technical field of biology, and discloses a D-dimer detection kit and a preparation method thereof. The kit comprises a buffer solution, D-dimer, a coagulation accelerator, a stabilizing agent, and an antiseptic; wherein the coagulation accelerator is polyvinylpyrrolidone (PVP). By adding polyvinylpyrrolidone (PVP) taken as a coagulation accelerator into the D-dimer detection kit, the combination between an antigen and an antibody is promoted to form a compound rapidly, at the same time, the dissociation of immune complex is inhibited, the precipitate appears rapidly, the detection speed is accelerated; moreover, the non-specific range is not enlarged, and the wide linear range of the D-dimer detection kit is maintained. Furthermore, PVP can fix D-dimer monoclonal antibodies on latex particles, the change of turbidity is specifically enhanced, and thus the detection sensitivity of the kit is improved.

The invention provides a high-efficiency specificity enriched hemagglutinin peptide HA mark recombinant protein immunoaffinity purification enriching column as well as a preparation method and an application thereof. The immunoaffinity purification enriching column provided by the invention comprises an activated agarose filler coupled with HA specificity polyclonal antibody which is subjected to immunoaffinity purification and a plastic column carrying the immunoaffinity filler. The immunoaffinity purification filler coupled HA specificity polyclonal antibody is extracted by using an immunoaffinity method; and the immunoaffinity purification enrichment column is obtained by carrying the immunoaffinity filler into a specific plastic column. The immunoaffinity purification enrichment column enriched HA mark recombinant protein is high in efficiency, strong in specificity and temperate in elution condition, can be repeatedly utilized and is an ideal material for separating, purifying and enriching the HA mark recombinant protein at present.

The present invention discloses a preparation method for a umami peptide affinity column. The method comprises the following steps: synthesizing receptor proteins T1R1 and T1R3; taking protein T1R1/T1R3 agarose gel as a solid phase support; coupling the protein T1R1/T1R3 on the support with umami peptide; and chemically bonding the coupled umami peptide with a stationary phase. The preparation method for the affinity column is simple and reusable; purification efficiency is improved, so that the sensitivity of the method is improved; theoretically the method is applicable to various sample matrices; and due to the protein receptors T1R1 / T1R3, the method has the good separation effect on the umami peptide, high coupled specificity, a large adsorption amount and can be used repeatedly.

The invention discloses a nanoscale single exosome separation method. The method comprises the following steps of fully diluting a cell culture solution containing exosome; adding a superparamagneticamino magnetic bead diluent, and carrying out low-frequency oscillation mixing until the nano magnetic beads are coupled with the exosome; transferring a coupling solution to a surface layer of a nanopore array microchip; placing the microchip on a shaking table, and oscillating in low speed and small angle, so that the coupling solution is uniformly distributed on the surface layer of the chip; placing a magnetic source on the lower surface of the microchip, and enabling a magnetic bead exosome couplet to enter a nanopore array by using a progressive magnetic adsorption mode; under the actionof larger magnetic force, maintaining stable adsorption of the magnetic bead exosome couplet, cleaning the upper surface of a substrate with a buffer solution, and standing to obtain the nano microchip containing the single exosome which can be directly used for subsequent detection. On the basis of the nanopore microarray microchip with the high-precision screening effect, high-purity single exosome can be obtained through magnetic bead coupling exosome and progressive magnetic adsorption.

The invention provides vomitoxin and a derivative aptamer affinity column and a preparation method and application thereof. The affinity column is prepared by the following steps: using N-hydroxysuccinimide modified agarose as a carrier, then carrying out covalent coupling on the carrier and a nucleic acid aptamer capable of high-affinity and high-specificity recognition of deoxynivalenol, deoxynivalenol-3-glucoside, 3-acetyl deoxynivalenol and 15-acetyl deoxynivalenol in a sample, and finally filling an affinity column with the coupled aptamer composite carrier. The affinity column is mainlyused for purification and decontamination of vomitoxin and its derivative in food, feeds, milk, food crops, traditional Chinese medicines and other various samples, which is beneficial to high performance liquid chromatography and rapid detection of toxins in the sample in later period. By one-time loading of the column, four types of toxins can be purified and enriched, the number of reuse can reach 15 times, recovery rate is 80% and above, the cost is reduced, and the sample pre-treatment efficiency of the sample is enhanced.

The invention provides an ofloxacin-hemocyanin coating antigen and a preparation method thereof. The structure of the ofloxacin-hemocyanin coating antigen is shown in the description. The preparation method of the ofloxacin-hemocyanin coating antigen includes the steps that C-chain derivatization is conducted on carboxyl of ofloxacin, activation is conducted on KLH at the same time, an acid-anhydride coupling method is adopted to synthesize the coating antigen, and the prepared coating antigen has the advantages of high coupling efficiency and reactogenicity. The invention further provides a test card for testing the ofloxacin, the test card is fast in detection and high in specificity and sensitivity, the detection limit of the test card can reach 0.06 ng/ml, and the requirements of rapid detection are met at the same time.

The invention provides a seafood product detection kit and a preparation method thereof, and belongs to the field of microbiological detection. The seafood product detection kit comprises anti-vibrioparahaemolyticus immunomagnetic beads and CdSe/ZnS-carboxylated polystyrene immunofluorescent microspheres. The magnetic beads provided by the invention are relatively high in surface carboxyl modification amount, the coupling rate of antibodies can be increased, and the capture rate of vibrio parahaemolyticus can be increased. According to the preparation method of the CdSe/ZnS-carboxylated polystyrene immunofluorescent microspheres, the cross-linking polymerization of the fluorescent microspheres is reduced, the dispersity is improved, the coupling rate of an antibody is improved, and the fluorescence intensity value is increased. The seafood product detection kit provided by the invention is used for detecting the vibrio parahaemolyticus and has relatively high sensitivity, accuracy andreliability.

The invention discloses a preparation method of an immunoaffinity column. The immunoaffinity column is a compound affinity column which is mainly prepared from three types of immunoaffinity columns through mixed packing in a volume ratio being 1:1:1, and the three types of immunoaffinity columns are prepared from three antibodies and a coupling matrix through coupling and wet packing. The column capacity of the immunoaffinity column is 2,500 ng, the immunoaffinity column is good in purification effect and can be repeatedly used three times, and average recovery rate is 67%-107%. The compound immunoaffinity column has good specific adsorption property for at least 12 sulfonamides including sulfaquinoxaline, sulfadiazine, sulfapyridine, sulfametoxydiazine, sulfadimidine, sulfamethoxazole, sulfafurazole, sulfadimethoxypyrimidine, sulfathiazole, sulfamerazine, sodium N-sulphanilamidate and sulfamonomethoxine. The immunoaffinity column which is high in coupling rate, specific adsorption property, column capacity and recovery rate is prepared, and a sensitive, rapid and simple analysis method capable of detecting residues of the 12 sulfonamides simultaneously is provided.

The invention provides a lactoferrin aptamer affinity column as well as a preparation method and application thereof. The affinity column takes agarose modified by N-hydroxysuccinimide as a carrier, anucleic acid aptamer capable of recognizing lactoferrin with high affinity and high specificity is covalently coupled with the carrier, and a solid-phase extraction column is filled with a specific affinity filler after coupling. The affinity column is mainly applied to specific recognition and enrichment purification of lactoferrin in a sample, the natural activity of lactoferrin can still be kept after purification, the method is rapid, simple, convenient and accurate, the affinity column can be repeatedly used, and the affinity column can be used for sample pretreatment before detection ofa large instrument and can also be used for separation and purification of lactoferrin. Wide application prospects are realized in the fields of cosmetics, food, animal production, medical treatmentand the like.

The invention provides a D-tag tagged recombinant protein immunoaffinity purification and enrichment column capable of efficiently and specifically enriching flag peptide tagged protein and a preparation method and application thereof. The provided immunoaffinity purification and enrichment column comprises an activated agarose filler coupled with an immunoaffinity-purified D-tag specific polyclonal antibody and a plastic column loaded with the immunoaffinity filler, wherein the D-tag specific polyclonal antibody coupled with the immunoaffinity filler is extracted by an immunoaffinity method. The immunoaffinity purification and enrichment column is prepared by loading the immunoaffinity filler into the special plastic column; and the immunoaffinity purification and enrichment column is high in efficiency and strong in specificity while enriching D-tag tagged recombinant protein, is mild in diluting conditions, can be repeatedly utilized and is an ideal material for separating, purifying and enriching the D-tag tagged recombinant protein.