43results about How to "Specific binding" patented technology

The invention belongs to the technical field of biomedicine and particularly relates to a CTLA-4 (cytotoxic T-lymphocyte-associated protein 4) nanoantibody and a preparation method and application thereof. The ability to bind with antigens can be improved or collaboratively enhanced through the CTLA-4 antibody. The CTLA-4 antibody has no complete antibody structure and lacks Fc end and Y-shaped structure; the antigens bound to the CTLA-4 antibody are not easily recognizable and can escape easily from being caught by the immune system. The technical means includes mainly: carrying out antigen-vector connecting; transferring to competent cells, and extracting recombinant plasmids; transfecting to host bacteria for expressing; immunizing camels with an antigen protein; collecting peripheral blood, extracting total RNA of a lymphocyte sample, inversely transcribing the total RNA into cDNA, performing amplifying to obtain a VHH library fragment, inserting the VHH library fragment into an expression vector, and transferring to the competent cells to construct an antibody immunization library; screening nanoantibodies in the antibody immunization library; verifying binding of the screened nanoantibodies with a CTLA-4 antigen.

The invention relates to an anti-CD38 nano-antibody, an encoding gene and an application. The anti-CD38 nano-antibody is following protein a) or b): a), protein consisting of an amino acid sequence shown as a sequence 2 in a sequence table; b), protein which is obtained by the amino acid sequence shown as the sequence 2 in the sequence table through substitution and / or deletion and / or addition ofone or more amino acids, related with specifically recognized CD38 and derived from a). The anti-CD38 nano-antibody can be efficiently and specifically bound with CD38, affinity is as high as 4 nM, large-scale production with low cost can be realized in an Escherichia coli expression system, and the anti-CD38 nano-antibody has significant application prospect in the fields of detection and pharmaceuticals.

The invention discloses a drug-loaded polymer vesicle with an asymmetric membrane structure, a preparation method and application in preparation of a drug for treating acute myeloid leukemia. An amphiphilic triblock polymer with polyaspartic acid PAsp, a targeted amphiphilic block polymer and a small molecule drug are assembled together to prepare the targeted small molecule drug-loaded polymer vesicle with the asymmetric membrane structure. The drug-loaded polymer vesicle disclosed by the invention has many unique advantages, including small size, simple and controllable preparation, reversible crosslinking, in-vivo stability, targeted delivery, high intracellular drug enrichment concentration, sensitive reduction, efficient killing of tumor cells, significant tumor growth inhibition effect and the like, and especially the drug-loaded vesicle disclosed by the invention has effective inhibition on both acute myeloid leukemia cell lines and patient cells. Therefore, the polymer vesicleis expected to become a simple and multifunctional nano platform for efficient and specific targeted delivery of drugs to tumor cells.

The invention belongs to the technical field of biomedicine and particularly relates to a PD-1 (programmed death 1) nanoantibody and a preparation method and application thereof. The ability to bind with antigens can be improved or collaboratively enhanced through the PD-1 nanoantibody. The PD-1 nanoantibody has no complete antibody structure and lacks Fc end and Y-shaped structure; the antigens bound to the PD-1 nanoantibody are not easily recognizable and can escape easily from being caught by the immune system. The technical means includes mainly: performing antigen-vector connecting; transferring to competent cells, and extracting recombinant plasmids; transfecting to host bacteria for expressing; immunizing camels with antigen proteins; collecting peripheral blood, extracting total RNA of a lymphocyte sample, reversely transcribing the total RNA into cDNA, performing amplifying to obtain a VHH library fragment, inserting the VHH library fragment into an expression vector, and transferring to the competent cells to construct an antibody immunization library; screening nanoantibodies from the antibody immunization library; verifying the binding of the screened nanoantibodies with a PD-1 antigen.

The invention provides sgRNA, a CREBRF point mutation-type Bama miniature pig constructed by the sgRNA, and application. The sgRNA comprises a nucleic acid sequence as shown in SEQ ID NO: 1 or SEQ IDNO: 2. According to the invention, a scheme of combining CRISPR / Cas9 with homologous recombination is adopted, point mutation of CREBRF gene is carried out, the designed gRNA has specific binding property on a target site near the 1370th site of the CEREBRF gene, the CRISPR / Cas9 can be guided to carry out specific recognition cutting near the target site, double-stranded DNA breakage is formed, homologous recombination repair with exogenous single-stranded DNA as a template is promoted, and therefore the point mutation is introduced. The mutant Bama miniature pig prepared by the method has a typical obesity phenotype and can be used as a model organism to be applied to the field of research and development of medicines for treating obesity and diabetes.

The invention belongs to the technical field of resistant substances from animals or humans, and discloses a TIM-3 nanobody as well as a preparation method and application thereof. The nanobody is TIM-3 nanobody, and the sequence is shown in SEQ ID NO:1; and a TIM-3 antigen is expressed by mammalian cell transient transfection, a nanobody library is screened for multiple times by using the TIM-3 antigen to obtain specific nanobody phage, and sequencing is performed to obtain a target fragment. An HEK293 cell line is used to express the antigen, a mammalian expression system is used to expresshuman protein to guarantee the original structure of the protein to the maximum extent to ensure that the protein has post-translational modification and specific modification, such as glycosylation of eukaryotic protein, so that the obtained protein has higher activity; the method guarantees the original structure and activity of the protein to the greatest extent; and the screened nanobody can efficiently and specifically bind to a target.

The invention provides a Helicobacter pylori antibody and its secreting cell line, preparation method and application. In the present invention, the ultrasonic supernatant of the cultured Helicobacter pylori protein is used as an immunogen to immunize mice, and the antibody 15A5 that can specifically bind to the Helicobacter pylori protein is obtained after screening, and the antibody is IgG2a subtype. This antibody detects infected H. pylori in tissue or serum.

The invention belongs to the technical field of soil protection, and in particular relates to a repair method for farmland soil heavy metal pollution. In this method, the heavy metal passivation agent is mixed evenly with the soil. After the maintenance, the first crop is planted, and then the hyperaccumulator plant is planted. At the same time, the heavy metal activator is applied, and then the passivation agent is applied. After the second crop is planted, the heavy metal passivation is applied first. The leaching agent can effectively reduce the risk of heavy metal leaching, and at the same time avoid the accumulation of heavy metals in crops, and then apply heavy metal activators to plant hyper-accumulator plants, absorb heavy metals, reduce the content of heavy metals in the soil, and prevent the accumulation of heavy metals in the soil in the area not covered by plant roots. Heavy metals keep the risk of leaching low due to the presence of heavy metal deactivators. The invention adopts the plant enrichment technology and the method of applying the heavy metal passivation agent, which can fundamentally remove the problem of heavy metals, and can passivate the heavy metals when planting crops, thereby improving the use efficiency of the soil.

The invention discloses a light-chain variable region and a heavy-chain variable region of an FMU-EPCAM-4F6 monoclonal antibody, wherein an amino acid sequence of the light-chain variable region of the FMU-EPCAM-4F6 monoclonal antibody is shown as SEQ ID NO.1, and an amino acid sequence of the heavy-chain variable region of the FMU-EPCAM-4F6 monoclonal antibody is shown as SEQ ID NO.2. In the invention, a group of mouse anti-human EPCAM monoclonal antibodies is prepared by a recombinant human EPCAM immune BALB / c mouse, hybridoma cell strains which can stably secrete high-affinity anti-human EPCAM monoclonal antibodies are screened, and the high-affinity anti-human EPCAM monoclonal antibodies are obtained by preparing ascitic fluid. The invention confirms the uniqueness of a gene sequence and a corresponding protein sequence as well as a CDR sequence and provides support for anti-body EPCAM chimeric or humanized gene engineering antibodies.

The present invention discloses light chain and heavy chain variable regions of an FMU-EPCAM(Epithelial Cell Adhesion Molecule)-2D7 monoclonal antibody, wherein an amino acid sequence of the light-chain variable region of the FMU-EPCAM-2D7 monoclonal antibody is disclosed as SEQ ID NO.1; and an amino acid sequence of the heavy-chain variable region is disclosed as SEQ ID NO.2. A group of mouse antihuman EPCAM monoclonal antibodies are prepared by recombinating human EPCAM immune BALB / c mousses, ascitic fluid is prepared by screening hybridoma cell lines which can stably secrete high-affinity antihuman EPCAM monoclonal antibodies, and the high-affinity antihuman EPCAM monoclonal antibodies can be obtained. The present invention ensures the uniqueness and a CDR sequence thereof of a gene sequence and a corresponding protein sequence and provides a support for antihuman EPCAM chimerisms or humanized genetic engineering antibodies.

The invention discloses a carbendazim monoclonal antibody, a preparation method and an application thereof. The carbendazim monoclonal antibody is prepared by a mouse hybridoma cell strain MC-3 with a preservation number of CGMCC N0.4575 and the preparation method is as follows: (1) introducing ninth amino acid residue of carbendazim into one amino group to obtain the carbendazim modified by the amino and coupling the carbendazim modified by the amino with a carrier protein to obtain a complete antigen A; (2) immunizing a mouse by the complete antigen A obtained in step (1), taking splenocyte of the immune mice to fuse with a mouse myeloma cell to screen out a positive hybridoma cell strain; extensively culturing the positive hybridoma cell strain and injecting the positive cell strain into a homologous mouse abdominal cavity to induce to generate ascitic fluid and to obtain more carbendazim monoclonal antibodies. The Carbendazim monoclonal antibody can be used for detecting the carbendazim.

The invention relates to a single-molecule protein detection chip and method based on electrochemiluminescence of an ultra-micro electrode array, and specifically discloses that the single-molecule protein detection chip has a substrate with an ITO electrode and a slanted microstructure with a three-dimensional concave on the ITO electrode. Well array; two ends in the lateral direction of the inclined micro well array are respectively provided with wires connected to the positive and negative electrodes of the power supply. The single-molecule protein detection chip of the invention is simple to prepare, the electrochemiluminescence detection system further amplifies the signal, and can realize the highly sensitive single-molecule detection of trace proteins.

The invention discloses application of a compact protein C-terminal fragment (CAF22) detection reagent in preparation of a renal tubule injury diagnostic kit and the renal tubule injury diagnostic kit, and provides a new biomarker for diagnosis of renal tubule injury by using the CAF22 as a diagnostic marker of the renal tubule injury. According to the invention, CAF22 detection is combined with other renal injury biomarkers, so that the accuracy and sensitivity of early diagnosis of renal tubule injury can be improved, and the kit has high application value in early diagnosis and treatment of renal injury.