1012 results about "Single chain" patented technology

The invention relates to the technical field of a preparation method of a type A recombinant botulinum toxin, in particular to a type A botulinum toxin obtained by adopting a gene recombination technology, and the amino acid sequence of the type A botulinum toxin is consistent with the sequence of the type A botulinum toxin prepared by extracting and purifying a wild strain Clostridium botulinum. According to the technical scheme, the single-chain protein of the botulinum toxin A sequentially comprises a botulinum toxin A light chain or a modified botulinum toxin A light chain, a connecting region and a botulinum toxin A heavy chain. According to the scheme, the high-quality recombinant botulinum toxin A is obtained after the single-chain protein is subjected to fermentation, crude purification, enzyme digestion and fine purification. According to the technical scheme, the problems of complex process, long period, high safety risk and the like in the existing process for preparing the botulinum toxin A from the wild bacterial strain clostridium botulinum can be solved, and meanwhile, the problem that the amino acid sequence consistent with that of the botulinum toxin A prepared from the wild bacterial strain clostridium botulinum cannot be obtained through the existing gene recombination technology is solved.

The present disclosure provides bispecific chimeric antigen receptors targeting CD20 and BCMA. The CAR may comprise an scFv targeting CD20 and an scFv targeting BCMA, a hinge region, a transmembrane domain, a co-stimulatory region, and a cytoplasmic signaling domain. The chimeric antigen receptors can be used to treat autoimmune disorders or cancer.

The invention relates to the field of gene therapy, in particular to an RBM25 gene epigenetics activation technology based on a CRISPR / dCas9-SunTag system and application of the RBM25 gene epigenetics activation technology in ischemic heart failure treatment, the RBM25 gene epigenetics activation technology comprises the following steps: (a) protein containing inactivated Cas9 (dCas9) fused with a plurality of GCN4 peptide repetitive sequences, the number of the GCN4 peptide repetitive sequences is 8-12, and the dCas9 contains D10A and H840A double mutations; (b) a protein containing fusion of an anti-GCN4 single-chain antibody fragment (scFv) and a TET1 catalytic structural domain; (c) at least one gRNA of a target RBM25 gene promoter region, wherein a target sequence of the gRNA is selected from a sequence as shown in SEQ ID NO: 1-3; and (d) a lipid nanoparticle (LNP) system for delivery of said (a), (b), and (c).

The invention relates to a biological preparation for improving quality of weever, the biological preparation comprises an injection and an edible agent, the injection comprises chemically modified chemically synthesized single-stranded small RNA, Rev-erbalpha protein and Per1 protein, and the edible agent comprises Clock core peptide 1, Clock core peptide 2 and taurine. Skeletal muscle development, liver lipid metabolism and generation of cell inflammatory factors of the weever are influenced by adding and injecting the feed, so that the quality of the weever is improved, intramuscular fat deposition is promoted, the taste and flavor of fish meat are improved, the energy reserve of the weever can be improved, and the culture cycle is shortened. The improvement of cell inflammatory factors shows that the disease resistance of the weever is improved, the survival rate in the breeding process is further improved, and the use amount of antibiotics is reduced. The method provides an important theoretical basis for improving fish meat quality in China and promoting high-quality, efficient and healthy development of aquaculture, and has a wide market prospect.

Disclosed is an immunity-inducing agent having excellent ease of production and high immunostimulatory activity, the immunity-inducing agent comprising, as an active component, a polynucleotide / peptide conjugate in which an antigenic peptide is covalently bound to a single-stranded polynucleotide or polynucleotide derivative. Also disclosed is a pharmaceutical composition comprising said immunity-inducing agent.

The invention discloses preparation of a human CD56 immune phage display antibody library, which comprises the following steps: S1, animal immunization: expressing recombinant hCD56 protein in escherichia coli, then immunizing a Balb / c mouse, and detecting immune titer; s2, splenocyte preparation: extracting RNA (Ribonucleic Acid) of immune mouse spleen cells, and performing reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid); s3, preparation of an initial bacteriophage display library: amplifying a heavy chain variable region and a light chain variable region by using a PCR technology, and then assembling to form scFv; constructing scFv phagocytids, electrically transforming the scFv phagocytids into TG1 competent cells, and constructing a bacterial library; s4, constructing a phage single-chain library of the hCD56: infecting the bacterial library by using a helper phage M13K07, and constructing a phage single-chain antibody library; and S5, elutriation and screening of a phage library: specifically enriching a phage single-chain antibody library, screening positive clones from the phage single-chain antibody library, and determining a sequence. According to the invention, the human hCD56 antibody scFv sequence fragment can be screened out, and the research and development period is shortened.

The invention discloses a recombinant fusion protein of a human lambda light-chain single-chain antibody, a DNA (deoxyribonucleic acid) molecule, a library building method and application, and the recombinant fusion protein of the human lambda light-chain single-chain antibody is the recombinant fusion protein formed by connecting the human lambda light-chain single-chain antibody, phage Piii protein and luciferase, the amino acid sequence of the recombinant fusion protein of the human lambda light-chain single-chain antibody is shown as SEQ ID NO: 5, the recombinant fusion protein library of the human lambda light-chain single-chain antibody has large capacity, the established single-chain antibody TG1 library is more than 1 * 10 < 9 > cfu / mL, positive clones capable of being combined with ligands can be screened and enriched for multiple rounds, and the recombinant fusion protein library has the characteristics of high sensitivity, high signal-to-noise ratio and the like.

The invention discloses a recyclable biochip stationary phase as well as a preparation method and application thereof, and belongs to the field of biochips. The preparation method of the biochip stationary phase comprises the following steps: (1) activating a glass slide by adopting plasma, and then putting the glass slide into a silane coupling agent solution to react, so that a silane coupling agent is covalently bound with the glass slide to obtain a functionally modified biochip; and (2) combining the 5'end modified single chain oligo with the functionally modified biochip to obtain the biochip stationary phase. The biochip stationary phase disclosed by the invention is low in cost, excellent in physicochemical property, good in stability and tolerance and capable of adapting to a complex environment; more importantly, the method can be recycled, the resource utilization rate is increased, the pollution is reduced, and the method has huge application potential in the leading technical field of DNA synthesis by an enzyme method and the like.

The invention discloses a hybridoma cell strain 1D6 as well as a monoclonal antibody and application thereof. The invention also discloses an application of the monoclonal antibody for resisting the Tilletia foetida in the detection of teliospore of the Tilletia foetida. The invention also discloses a single base sequence DNA single chain modified Tilletia foetida monoclonal antibody. The invention also discloses a detection kit. The monoclonal antibody as well as the marker and the kit thereof show high specific recognition capability on Tilletia foetida. The method for detecting the single base by mass spectrometry has a signal amplification effect and is simple and convenient in detection operation, so that the sensitivity and the detection efficiency of the method are remarkably improved. With the progress and continuous popularization and application of the mass spectrum technology, the technology has wide application potential and prospect.

Provided herein are methods of treating a cancer in a subject, wherein the cancer expresses a Ro52 antigen, comprising administering an anti-Ro52 binding agent to the subject, thereby treating the cancer in the subject. In some embodiments, the Ro52 antigen is expressed on a cancer cell surface. In some embodiments, the anti-Ro52 binding agent comprises a Fab, Fab', F(ab')2, Fahl-SH, Fv, diabody, linear antibody or single-chain variable fragment (scFv). In some embodiments, the anti-Ro52 binding agent is a humanized or chimeric antibody. In some embodiments, the anti-Ro52 binding agent is conjugated to a therapeutic agent.

The invention discloses a double-target chimeric antigen receptor for co-expression of CD47 and IL-15, a CAR-T cell and application of the CAR-T cell, and belongs to the technical field of genetic engineering, the double-target chimeric antigen receptor comprises a single-chain antibody scFv-EGFRv III, a single-chain antibody scFv-VEGF, CD47 and IL-15; the nucleotide artificial sequence of the single-chain antibody scFv-EGFRv III is as shown in SEQ ID NO. 6; the nucleotide artificial sequence of the single-chain antibody scFv-VEGF is as shown in SEQ ID NO. 9. According to the application disclosed by the invention, by performing sequence optimization on targets EGFRvIII and VEGF, the anti-tumor effect of CAR-T cells in glioblastoma can be improved, and by integrating immune checkpoint molecules CD47 and interleukin-15, the immunosuppression function of a CD47-SIRP alpha signal channel can be locally exerted.

The invention belongs to the technical field of biological medicine and molecular biology, and particularly relates to a single-chain antibody targeting NRP1, a CAR-T cell and application of the single-chain antibody and the CAR-T cell in fibrosis treatment. Four types of high-affinity single-chain antibodies targeting mouse NRP1 antigens are obtained by immunizing mice, the single-chain antibodies are applied to second-generation CAR, T cells of mouse spleen sources are infected after virus packaging, and CAR-T cells are constructed. The CAR-T cells of the targeted NRP1 can be used for effectively killing NRP1 positive cells. In-vivo experiments show that the CAR-T cells can inhibit the growth of NRP1 positive tumor cells. The CAR-T cell can improve the mouse liver fibrosis condition, collagen in the mouse liver is reduced through treatment, and the molecular level of fibrosis markers SMA and NRP1 is reduced, it is indicated that the CAR-T cell recognizes and eliminates activated HSCs through targeting NRP1, and the fibrosis degree is reduced.

Provided herein are chimeric receptors, including chimeric antigen receptors (CARs), comprising BCMA-binding molecules, including anti-BCMA antibodies and antigen-binding fragments thereof, including heavy chain variable (VH) regions and single-chain antibody fragments, and encoding polynucleotides. In some embodiments, the anti-BCMA chimeric receptors specifically bind to BCMA. Among the anti-BCMA-binding molecules are human antibodies, including those that compete for binding to BCMA with reference antibodies, including a non-human reference antibody. Also provided are genetically engineered cells expressing the CARs and uses thereof including in adoptive cell therapy.

Disclosed are a preparation method and application of a single-chain antibody fragment targeting SARS-COV-2 nucleocapsid protein, aiming to provide a single-chain antibody fragment with high affinity and strong specificity. The present disclosure obtains three single-chain antibody fragments targeting SARS-COV-2 nucleocapsid protein through construction of a nanobody phage library and phage screening technology. The single-chain antibody fragment consists of a heavy chain variable region, a 15aa connecting peptide, and a light chain variable region connected in sequence, and has an amino acid sequence shown in SEQ ID NO: 1-SEQ ID NO: 3. The single-chain antibody fragment in the present disclosure has excellent binding performance and specificity with the SARS-COV-2 nucleocapsid protein, can be applied to a variety of immunoassay platforms of the SARS-COV-2, and has broad application prospects in the field of SARS-COV-2 detection.

The invention provides a reagent for detecting avian leukosis virus, a nano-enzyme immunochromatography test strip and application of the nano-enzyme immunochromatography test strip, and belongs to the technical field of immunology. Aiming at the problems that the existing avian leukosis virus detection method is complicated in operation, time-consuming or strong in equipment dependence, the reagent comprises a capture antibody and a detection antibody; a heavy chain variable region of the monoclonal antibody A comprises an amino acid sequence as shown in SEQ ID NO: 1, and a light chain variable region of the monoclonal antibody A comprises an amino acid sequence as shown in SEQ ID NO: 2; a heavy chain variable region of the monoclonal antibody B comprises an amino acid sequence as shown in SEQ ID NO: 5, and a light chain variable region of the monoclonal antibody B comprises an amino acid sequence as shown in SEQ ID NO: 6; the antigen binding fragment is a Fab fragment, an F (ab) 2 fragment or a single-chain Fv fragment. The reagent is mainly used for detecting avian leukosis virus, and is suitable for detecting samples such as cloaca swab, meconium, egg white or serum of poultry.

Provided herein are antibodies and antibody fragments that bind to human TNFRSF25. The antibodies may be monoclonal and / or biparatopic antibodies and / or single- chain fragment variable (scFv) antibodies. Methods of treating or preventing diseases or disorders associated with inflammation and / or autoimmunity are provided, comprising administering to a patient in need thereof an effective amount of a human TNFRSF25- binding antibody.

This disclosure relates methods for expanding γδ T cells. More specifically, the disclosure relates to methods of expanding γδ T cells that have an eliminated or reduced expression of endogenous MHC Class I molecules, by expressing a single-chain fusion HLA Class I protein. In some embodiments, the endogenous B2M gene of the γδ T cells is disrupted. In some embodiments, the single-chain fusion HLA Class I protein includes at least a portion of B2M protein and at least a portion of HLA-E heavy chain.

The present invention is directed to a humanized monoclonal anti-human EpCAM antibody, such as a single-chain variable fragment (scFv), comprising VH having the amino acid of SEQ ID NO: 2 and VL having the amino acid of SEQ ID NO: 4. The present invention is also directed to a chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) a single-chain variable fragment (scFv) of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain.

The preparation and evaluation method comprises the following steps: 1, extracting a heavy-chain DNA fragment and a light-chain DNA fragment, carrying out recombinant transformation by using an overlapping PCR method to construct an scFv gene, 2, constructing an expression vector, culturing the expression vector, carrying out sequencing identification to obtain an scFv bacterial solution, recombining the scFv bacterial solution with a mouse Fc fragment, and identifying a sequence to obtain an MN8 bacterial solution, the method comprises the following steps: 1, preparing an MN8 bacterial liquid, 2, extracting the MN8 bacterial liquid, 3, carrying out inoculated culture on the MN8 bacterial liquid to obtain an MN8 genetically engineered antibody bacterial liquid, 4, extracting MN8 genetically engineered antibody bacterial liquid plasmids and carrying out transfection expression purification, and 5, carrying out antibody titer and thermal stability determination on the MN8 genetically engineered antibody. The scFv single-chain antibody obtained by recombination according to the method has the advantages of small molecular weight, high penetrability and the like, the stability and the sensitivity of an immunoassay method of the scolosporins can be improved, and a certain basis is provided for rapid determination of the scolosporins.

The invention provides a soybean peptide with an effect of preventing and / or treating chronic nephritis as well as a preparation method and application of the soybean peptide, and belongs to the technical field of bioactive peptides. According to the preparation method of the soybean peptide mixture provided by the invention, a three-step enzymolysis mode is adopted, the molecular weight of soybean peptide is controlled to be 1000 Daltons or below, micromolecular oligopeptide is used as a main component, and the prepared soybean peptide mixture is low in sodium, phosphorus and potassium content and capable of improving the nutritional status and muscle loss condition of nephritis patients. The prepared soybean peptide mixture is further separated and purified, the purified soybean peptide mixture, the single-chain polypeptide S1 and the single-chain polypeptide S2 are obtained, the content of uric acid and creatinine can be effectively reduced, inflammation can be inhibited, and the soybean peptide is expected to be developed into a novel polypeptide medicine for preventing and / or treating chronic kidney diseases.

The invention relates to the field of biotechnology and biological breeding, and particularly relates to a method for constructing a single-chain guide RNA (sgRNA) vector for delivering CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) by utilizing a cucumber mosaic virus (CMV), carrying out site-specific editing on a target gene in a Cas9 transgenic plant, causing site-specific mutagenesis of plant genome DNA (Deoxyribose Nucleic Acid) by the vector to be inherited to a progeny, and not carrying the CMV in the progeny. The invention discloses a construction method of a vector pCB301R3-sgPDS and an application of the vector pCB301R3-sgPDS. The pCB301R3-sgPDS is a genetic gene editing vector based on mediation of a cucumber mosaic virus. According to the invention, sgRNA is delivered through CMV for the first time, so that fixed-point editing and seed transmission of plant target genes are efficiently realized in crops.

The invention relates to an application of a macrophage expressing an anti-uPAR chimeric antigen receptor in preparation of an anti-aging drug, and discloses a new application of the macrophage (CAR-M) expressing the anti-uPAR chimeric antigen receptor in senescence diseases, and the mechanism is that uPAR positive cells are specifically recognized and apoptosis of the uPAR positive cells is induced by targeting senescence cell surface antigen uPAR. According to the uPAR-CAR-M, a humanized anti-uPAR single-chain antibody is used as a chimeric antigen receptor recognition region, so that the chimeric antigen receptor has high specificity, and the chimeric antigen receptor is used for promoting the functions of macrophages by using a CD8 alpha transmembrane structural domain, a CD28 costimulatory molecule and a Megf10 sequence, so that a better killing effect is given to the CAR-M. The problems of inflammatory microenvironment caused by abnormal aggregation of senescence cells, chronic tissue damage and senescence-related disease treatment disorder are solved, and the application value for treating senescence-related diseases (such as osteoarthritis and the like) expressing uPAR is important.

The invention belongs to the technical field of biomedical detection, and discloses a nucleic acid aptamer, a nucleic acid aptamer derivative and application thereof. An aptamer DNA long single chain targeting DLL3 is screened through an exponential enrichment systematic evolution (SELEX) technology, sequence optimization is performed on the aptamer DNA long single chain to obtain another two short chains, the aptamer can specifically target tumor cell surface DLL3, and the aptamer and small cell lung cancer cells SHP77, H69 and the like with high expression of the DLL3 have a specific targeting recognition effect. The invention also discloses screening and optimization of the nucleic acid aptamer. The nucleic acid aptamer has the advantages of being high in affinity and specificity, free of immunogenicity, simple and stable in chemical synthesis, easy to store and mark and the like.

The invention discloses an antigen binding fragment for resisting a sarbecovirus coronavirus, the antigen binding fragment comprises a complementarity determining region, the complementarity determining region comprises a CDR3, and the amino acid sequence of the CDR3 is shown as SEQ ID NO.3. The invention also discloses an antigen binding fragment for resisting the sarbecovirus coronavirus. The invention also discloses an antibody which comprises the antigen binding fragment. The invention also discloses a biological material related to the antigen binding fragment or the antibody, a method for preparing a single-chain antibody, a detection or diagnosis reagent and a pharmaceutical composition. The antigen binding protein and the antibody provided by the invention are broad-spectrum antibodies aiming at coronavirus, not only can effectively neutralize various variants, but also have high affinity and lasting effectiveness, can improve the ability of resisting newly appearing sbecovirus coronavirus strains, and are beneficial to improving the treatment effect and reducing the severe case rate.

The invention provides a compound for releasing a urine fluorescent biomarker under induction of mitochondrial DNA mutation as well as a preparation method and application of the compound, and belongs to the technical field of medicines. The compound disclosed by the invention is prepared by the following steps: complementarily pairing a hybrid nucleic acid nano-carrier containing single-stranded DNA (Deoxyribose Nucleic Acid), an annular fluorescent reporter molecule and a Cas12a / crRNA compound through bases of DNA or RNA, and then coating polyethyleneimine and hyaluronic acid. A CRISPR / Cas12a system is selected as a converter of tumor specific mitochondrial DNA mutation information and urine fluorescence signals, a nano delivery system which responds to mtDNA single base mutation and is used for tumor progress monitoring and early metastasis warning is designed and synthesized, and the nano delivery system has a good application prospect in tumor progress monitoring and early metastasis sensitive detection.

The invention relates to the technical field of gene engineering, in particular to application of wheat TaLNUE protein in regulation and control of low nitrogen stress tolerance of plants. According to the invention, a single-chain guide RNA of a target wheat TaLNUE gene is designed, an expression vector of the TaLNUE gene is knocked out, and the expression vector is transformed into wild type wheat, so that transgenic wheat with the function inhibited by the TaLNUE gene is obtained. Researches find that the spike length and the spikelet number of wheat under low nitrogen stress can be increased by inhibiting the expression of the wheat TaLNUE protein, so that the single plant yield and the total yield of the wheat under low nitrogen stress are increased, and the nitrogen absorption efficiency and the nitrogen utilization efficiency of the wheat under low nitrogen stress are improved. According to the technical scheme, a feasible method is provided for improving the wheat yield under low nitrogen stress by utilizing a genetic engineering technology.

The present invention is directed to a monoclonal anti-human CS1 clone 7A8D5 antibody or a single-chain variable fragment (scFv), comprising VH having the amino acid of SEQ ID NO: 4 and VL having the amino acid of SEQ ID NO: 5. The present invention is also directed to a chimeric antigen receptor fusion protein comprising from N-terminus to C-terminus: (i) CS1 scFv of the present invention, (ii) a transmembrane domain, (iii) at least one co-stimulatory domains, and (iv) an activating domain.

Provided are a CAR library used to screen scFvs that can be functional in CAR-T cells, and an scFv manufacturing method in which the CAR library is used. A chimeric antigen receptor (CAR) library of the present invention includes nucleic acids coding for first CARs. Each of the first CARs includes a first antigen-binding domain, a first transmembrane domain, and a first intracellular signaling domain. The first antigen-binding domain includes a first single-chain antibody (scFv) to be screened for the ability to bind to a target antigen. The first scFv includes a first heavy-chain variable region and a first light-chain variable region. The first heavy-chain variable region and the first light-chain variable region meet a predetermined condition.

The present invention provides a combined chimeric antigen receptor targeting CD19 and CD20 and application thereof. Specifically, the present invention provides a combined chimeric antigen receptor targeting CD19 and CD20, which comprises a scFv targeting CD19 and CD20, a hinge region, a transmembrane region, and an intracellular signaling domain. The present invention provides a nucleic acid molecule encoding the chimeric antigen receptor and a corresponding expression vector, a CAR-T cell, and applications thereof. The experimental results show that the chimeric antigen receptor provided by the present invention shows extremely high killing ability against tumor cells. The chimeric antigen receptor of the present invention targets CD19 and / or CD20 positive cells and can be used to treat CD19 and / or CD20 positive B-cell lymphoma, leukemia and other diseases.

The invention relates to the technical field of biological medicines, in particular to a recombinant oncolytic virus for expressing a bispecific or trispecific single-chain antibody and application of the recombinant oncolytic virus. The recombinant oncolytic virus comprises an antibody gene integrated into a virus skeleton of the recombinant oncolytic virus; the antibody gene comprises a target antibody gene targeting tumor cells and at least one antibody gene targeting immune cells. When the antibody gene comprises a target antibody gene targeting tumor cells and an antibody gene targeting immune cells, the antibody gene is a bispecific antibody; when the antibody gene comprises a target antibody gene targeting tumor cells and two antibody genes targeting immune cells, the antibody gene is a trispecific antibody. According to the present invention, the coding gene having the bispecific single-chain antibody or the trispecific single-chain antibody is integrated in the modified oncolytic virus, such that the gene of the bispecific single-chain antibody or the trispecific single-chain antibody can be expressed in vivo so as to further improve the tumor cell treatment effect.