15 results about "Ribosome display" patented technology

The invention discloses a double-epitope alpaca source nano antibody and application thereof, and belongs to the field of biological medicine. Aiming at different epitopes of MERS-CoV, specific screening is carried out by immunizing alpaca and utilizing a ribosome display nano antibody library, and VHH epitopes and molecular docking are identified by utilizing a pseudovirus neutralization experiment, a competitive BLI experiment and a phage random peptide library, so that two antibodies 2-77 and 4-58 aiming at different binding sites of MERS-CoV RBD are obtained. And the double-epitope alpaca source nano antibody 2F4 which is good in neutralizing activity, high in expression quantity and stable is constructed from the two strains of nano antibodies. A focus formation test and a mouse in-vivo experiment prove that the constructed double-epitope alpaca source nano antibody 2F4 can effectively neutralize the MERS-CoV and effectively protect animals, and a thought and a technical support are provided for exploring brand-new antigen epitopes and designing therapeutic antibody drugs for the MERS-CoV.

The invention belongs to the technical field of antibody library construction, and particularly relates to a synthetic nano library for ribosome display screening and construction and application thereof. Aiming at the problems that the existing nano antibody acquisition depends on animal immunity, the diversity and screening efficiency of a synthetic library are insufficient and the like, the invention discloses a synthetic nano antibody library suitable for ribosome display and a construction method of the synthetic nano antibody library, and the synthetic nano antibody library suitable for ribosome display is constructed by combining a trinucleotide primer synthesis and Type IIS enzyme digestion connection technology through a consensus skeleton design and a CDR region differentiation randomization strategy. The library stability and the effective diversity are improved. The library does not need to immunize animals, is short in period and low in cost, is compatible with toxic antigens, can be screened through combination of ribosome and bacteriophage, can efficiently obtain nano-mole-level affinity nano-antibodies, can also assist structure research of a cryoelectron microscope, has the advantages of being complete in library, efficient in screening, low in sequence redundancy and the like, and provides powerful support for research and development of biological medicines.

The invention discloses a nano antibody capable of specifically recognizing staphylococcus aureus as well as a preparation method and application of the nano antibody. A ribosome display technology is adopted, a nano antibody pair with a good pairing effect is screened out through construction of a nano antibody display library, three rounds of high-throughput screening and ELISA verification, and the nano antibody pair is used for in-vivo detection of actual pathogenic bacteria in combination with an immunochromatography technology. The nano antibody disclosed by the invention shows high affinity, high specificity and achievable detection limit when being combined with staphylococcus aureus, has the advantages of good stability, low cost and the like compared with a monoclonal antibody, can be produced on a large scale, and has a wide application prospect in the field of staphylococcus aureus detection and analysis.

The invention discloses a nano antibody capable of specifically recognizing staphylococcus aureus as well as a preparation method and application of the nano antibody. A ribosome display technology is adopted, a nano antibody pair with a good pairing effect is screened out through construction of a nano antibody display library, three rounds of high-throughput screening and ELISA verification, and the nano antibody pair is used for in-vivo detection of actual pathogenic bacteria in combination with an immunochromatography technology. The nano antibody disclosed by the invention shows high affinity, high specificity and achievable detection limit when being combined with staphylococcus aureus, has the advantages of good stability, low cost and the like compared with a monoclonal antibody, can be produced on a large scale, and has a wide application prospect in the field of staphylococcus aureus detection and analysis.

The application discloses a dual-epitope llama-derived nanobody and application thereof, and belongs to the field of biological medicine. The application is aimed at different epitopes of MERS-CoV, and two strains of antibodies 2-77 and 4-58 aiming at different binding sites of MERS-CoV RBD are obtained by immunizing a llama, performing specific screening on a ribosome display nanobody library, performing a pseudovirus neutralization experiment, performing a competitive BLI experiment, identifying VHH epitopes and molecular docking by using a phage random peptide library, and constructing a dual-epitope llama-derived nanobody 2F4 with good neutralization activity, high expression and stability by using the two strains of nanobodies. It is proved by a lesion formation test and a mouse in-vivo experiment that the dual-epitope llama-derived nanobody 2F4 constructed by the application can effectively neutralize MERS-CoV and effectively protect animals, and the application provides a train of thought and technical support for exploring new antigen epitopes of MERS-CoV and designing a therapeutic antibody drug.

The invention discloses a nano antibody capable of specifically recognizing staphylococcus aureus as well as a preparation method and application of the nano antibody. A ribosome display technology is adopted, a nano antibody pair with a good pairing effect is screened out through construction of a nano antibody display library, three rounds of high-throughput screening and ELISA verification, and the nano antibody pair is used for in-vivo detection of actual pathogenic bacteria in combination with an immunochromatography technology. The nano antibody disclosed by the invention shows high affinity, high specificity and achievable detection limit when being combined with staphylococcus aureus, has the advantages of good stability, low cost and the like compared with a monoclonal antibody, can be produced on a large scale, and has a wide application prospect in the field of staphylococcus aureus detection and analysis.

The application discloses a nano antibody specifically recognizing staphylococcus aureus and a preparation method and application thereof. The ribosome display technology is adopted to screen a pair of nano antibodies with good pairing effect through construction of a nano antibody display library, three rounds of high-throughput screening and ELISA verification, and the nano antibody is combined with an immunochromatography technology to be used for in-vivo detection of actual pathogenic bacteria. The nano antibody disclosed by the application shows high affinity and high specificity when being combined with the staphylococcus aureus, can realize a detection limit, and has the advantages of good stability and low cost compared with a monoclonal antibody, can be produced on a large scale, and has a wide application prospect in the field of staphylococcus aureus detection and analysis.

This invention provides a nanobody targeting canine OSMRβ receptor M and its applications, belonging to the field of nanobody preparation technology. This invention achieves recombinant expression and purification of the canine OSMRβ receptor, and obtains the targeting nanobody HX016 based on ribosome display technology. Enzyme-linked immunosorbent assay (ELISA) confirms that the nanobody HX016-cFc specifically binds to canine OSMRβ. SPR technology confirms that the nanobody HX016-cFc exhibits good affinity for canine OSMRβ. Cell-level experiments show that the nanobody HX016-cFc can effectively bind to canine OSMRβ and significantly inhibit IL-31-induced downstream signaling pathway activation in a dose-dependent manner, thereby effectively intervening in the IL-31-driven itching and inflammatory response in canine atopic dermatitis at the mechanistic level. A mutated portion of HX016 also possesses the corresponding ability to bind to the canine OSMRβ antigen.

The invention discloses a nano antibody capable of specifically recognizing staphylococcus aureus as well as a preparation method and application of the nano antibody. A ribosome display technology is adopted, a nano antibody pair with a good pairing effect is screened out through construction of a nano antibody display library, three rounds of high-throughput screening and ELISA verification, and the nano antibody pair is used for in-vivo detection of actual pathogenic bacteria in combination with an immunochromatography technology. The nano antibody disclosed by the invention shows high affinity, high specificity and achievable detection limit when being combined with staphylococcus aureus, has the advantages of good stability, low cost and the like compared with a monoclonal antibody, can be produced on a large scale, and has a wide application prospect in the field of staphylococcus aureus detection and analysis.

The invention discloses a nano antibody capable of specifically recognizing staphylococcus aureus as well as a preparation method and application of the nano antibody. A ribosome display technology is adopted, a nano antibody pair with a good pairing effect is screened out through construction of a nano antibody display library, three rounds of high-throughput screening and ELISA verification, and the nano antibody pair is used for in-vivo detection of actual pathogenic bacteria in combination with an immunochromatography technology. The nano antibody disclosed by the invention shows high affinity, high specificity and achievable detection limit when being combined with staphylococcus aureus, has the advantages of good stability, low cost and the like compared with a monoclonal antibody, can be produced on a large scale, and has a wide application prospect in the field of staphylococcus aureus detection and analysis.

The invention discloses a nano antibody targeting a brucella surface antigen and application of the nano antibody, and belongs to the technical field of biological medicines. The amino acid sequence of the nano antibody targeting the Brucella surface antigen is as shown in SEQ ID NO. 1. The method comprises the following steps: firstly, screening brucella surface antigen nano antibodies by virtue of a ribosome display technology, then constructing a recombinant expression vector, carrying out protein purification on all the nano antibodies which are correctly sequenced, carrying out affinity identification on all the nano antibodies by virtue of a brucella S2 strain after all the nano antibodies are completely purified, and selecting the nano antibody with the strongest affinity. A cell experiment and an animal experiment prove that the Brucella load in RAW264.7 macrophages treated by the nano antibody and the Brucella load in a mouse body are obviously reduced compared with a control group, and the nano antibody is proved to be capable of reducing Brucella in infected organisms and cells. The limitation that a traditional antibody cannot target intracellular bacteria is overcome, and a core material is provided for developing novel anti-brucella drugs.

The present invention discloses a nanobody specifically recognizing Staphylococcus aureus, and a preparation method and application thereof. The present invention adopts ribosome display technology, screens out a pair of nanobody pairs with good pairing effect through constructing a nanobody display library, three rounds of high-throughput screening and ELISA verification, and combines immunochromatography technology for in vivo detection of actual pathogenic bacteria. The nanobodies disclosed by the present invention show high affinity and high specificity when binding to Staphylococcus aureus, can achieve a detection limit, and have advantages such as good stability and low cost compared with monoclonal antibodies, can be mass-produced, and have broad application prospects in the field of Staphylococcus aureus detection and analysis.

The invention discloses a cell-free protein synthesis system for protein and polypeptide screening, and belongs to the technical field of biology. According to the escherichia coli engineering bacterium for the cell-free protein synthesis system, ssrA, arfB and smrB genes of the escherichia coli engineering bacterium are knocked out through gene editing, and proteins coded by the escherichia coli engineering bacterium are removed from the gene level; protease degradation tags are added to the C ends of prfA, prfB, prfC, pth, frr and arfA genes, and proteins coded by the proteins are removed from the protein level. The cell-free protein expression system prepared by using the engineered escherichia coli can completely remove proteins which influence the stability of the ternary complex and are unfavorable for ribosome display, can significantly improve the stability of the ternary complex, can perform protein synthesis under the condition of 30 DEG C, and can be used for screening proteins and polypeptides.

The application discloses a nano antibody specifically recognizing staphylococcus aureus and a preparation method and application thereof. The ribosome display technology is adopted to screen a pair of nano antibody with good pairing effect through construction of a nano antibody display library, three rounds of high-throughput screening and ELISA verification, and the nano antibody is combined with an immunochromatography technology to be used for in-vivo detection of actual pathogenic bacteria. The nano antibody disclosed by the application shows high affinity and high specificity when being combined with the staphylococcus aureus, can realize a detection limit, and has the advantages of good stability and low cost compared with a monoclonal antibody, can be produced on a large scale, and has a wide application prospect in the field of staphylococcus aureus detection and analysis.

The application discloses a nano antibody specifically recognizing staphylococcus aureus and a preparation method and application thereof. The ribosome display technology is adopted to screen a pair of nano antibodies with good pairing effect through construction of a nano antibody display library, three rounds of high-throughput screening and ELISA verification, and the nano antibody is combined with an immunochromatography technology to be used for in-vivo detection of actual pathogenic bacteria. The nano antibody disclosed by the application shows high affinity and high specificity when being combined with the staphylococcus aureus, can realize a detection limit, and has the advantages of good stability and low cost compared with a monoclonal antibody, can be produced on a large scale, and has a wide application prospect in the field of staphylococcus aureus detection and analysis.