69results about How to "Weak immunogenicity" patented technology

The invention discloses a method for inhibiting HIV-1 infectious agent from infecting primary lymphocyte by utilizing CRISPR/Cas9 and relates to genetic engineering and an anti-virus infection technology. According to the method, by utilizing the latest CRISPR/Cas9 nuclease system, through the combination of Ad5F35 mosaic type adenovirus vectors of efficient targeted primary T lymphocyte, by editing CCR5 expression of human CD4+T lymphocyte, HIV-1 virus infection is effectively inhibited. According to the method, the CRISPR/Cas9 system is packaged by utilizing the Ad5F35 type adenovirus vectors, the advantages of the CRISPR/Cas9 system and the Ad5F35 type adenovirus vectors are organically combined, the novel method for resisting HIV-1 virus infection is provided, and the method has the potential for being applied for HIV-1 gene treatment research; meanwhile, the Ad5F35 type adenovirus vectors carrying with the CRISPR/Cas9 system can be used for conducting targeted editing to other genes in the primary lymphocyte or gene editing research of other hematopoietic system cells.

The invention relates to a chimeric antigen receptor targeting BCMA, which comprises an extracellular domain capable of binding to an antigen, a transmembrane domain and at least one intracellular domain, wherein the extracellular domain is an anti-BCMA single domain antibody; wherein the amino acid sequence of the anti-BCMA single domain antibody is selected from: (a) an amino acid sequence as shown in SEQ ID NO. 1; or (b) a variant which is formed by substitution, addition or deletion of one or more amino acids in the amino acid sequence shown in SEQ ID NO. 1 and which is capable of specifically binding to the chimeric antigen receptor and has a function of binding to BCMA and inducing T cell signaling conduction. The chimeric antigen receptor has smaller clinical side effects and highersafety, can effectively reduce solid tumor focus and effectively improve the treatment effect of tumors.

The invention relates to the technical field of biology, in particular to a nano-antibody GN1 specifically combined with a GPC3 protein. The nano-antibody GN1 is composed of a variable region of a heavy-chain antibody, the variable region of the heavy-chain antibody comprises an antigenic determinant complementary region and a skeleton region, the skeleton region is selected from the group consisting of FR1, FR2, FR3 and FR4 and homologous sequences thereof, the antigenic determinant complementary region is selected from the group consisting of CDR1, CDR2 and CDR3 and homologous sequences thereof, the amino acid sequences of the CDR1 to the CDR3 are shown in the formulas of SEQ ID NO. 1 to SEQ ID NO. 3, the amino acid sequences of the FR1-4 are shown in formulas of SEQ ID NO. 4-7, the amino acid sequence of the antibody is shown in the formula of SEQ ID NO.8, and the nucleotide sequence for encoding the amino acid is shown in the formula of SEQ ID NO.9. The nano-antibody GN1 can be specifically combined with hepatoma carcinoma cells highly expressing the GPC3 protein to inhibit hepatoma carcinoma cell proliferation. The amino acid sequence of the nano-antibody GN1 or the nucleotidesequence of the nano-antibody GN1 or the recombinant plasmid containing the nucleotide sequence of the nano-antibody GN1 or the recombinant cell containing the recombinant plasmid containing the nucleotide sequence of the nano-antibody GN1 can be applied to research and development of diagnostic reagents and drugs for treating liver cancer.

The invention provides a kind of extract with its molecular weight being lower than 5800 dalton from hirudo and/ or lumbricus, the preparation method, medicine compound and its application. The mentioned extract is better than current hirudo and/ or lumbricus extract in terms of visible foreign matter and insoluble micro-particle and product safety. The product in this invention can be used to produce medicine for preventing and treating thrombus relevant diseases.

The invention provides an application of umbilical cord mesenchymal stem cells in preparation of formulation for treating lupus erythematosus, in particular to an application in preparation of formulation for treating recurrent systemic lupus erythematosus. The invention further provides a method of preparing umbilical cord mesenchymal stem cells of formulation capable of being used for treating the lupus erythematosus and a formulation composition having the stem cells. The umbilical cord mesenchymal stem cells prepared by the invention have the advantages of low immunoreaction risk, high activity, low residue, and the like, and the preparation cost is low.

The invention provides a stem cell injection for treating osteoarthritis cartilage defect and preparation and application thereof. The injection comprises fifth-generation umbilical cord mesenchymal stem cells and a liquid solvent, wherein the fifth generation umbilical cord mesenchymal stem cells are suspended in the liquid solvent, and the concentration is (2.0-3.0)x10<7> cells/ 4.5-5.0ml of the liquid solvent. The liquid solvent is normal saline, or autologous platelet-rich plasma, or a mixed solution of normal saline and human albumin, or a mixed solution of normal saline and autologous platelet-rich plasma. The human albumin accounts for 5-8% in volume by percentage. The normal saline accounts for 10% or below in volume by percentage. The injection is administered by intravenous injection or bone joint cavity local injection or the combination of the two, the cell injection amount of the intravenous injection is (1.0-1.5)x10<8> cells/ one time, and the cell injection amount of the bone joint cavity injection is (2.0-3.0)x10<7> cells/ one time/ one knee. The effect of the stem cell injection for treating the integrity degree of osteoarthritis cartilage is obviously superior to that of a traditional hyaluronic acid (HA) method.

The invention belongs to the field of molecular biology, and particularly relates to an EpCAM single domain antibody G7 and a nucleotide sequence for encoding the EpCAM single domain antibody G7. By utilizing a phage display technology, a single domain heavy chain antibody library specifically bound with a target antigen CD326 is screened from a camel-source immunizing single domain heavy chain antibody library, and a coding sequence of a positive clone is further guided into a prokaryotic expression vector, so that the high-efficiently expressed EpCAM single domain antibody G7 is obtained. Therefore, a basis is provided for clinic targeted immunological therapy of CD326.

The invention aims to provide polypeptide with immunomodulatory effects, namely undecapeptide which has immunomodulatory effects and is separated from bursa of Fabricius. The amino acid sequence of the polypeptide is SEQ ID NO:1. The bursa of Fabricius undecapeptide separated from a bursa of Fabricius extract product has a simple structure, has no toxic or side effect, and has poor immunogenicity. The polypeptide can be obtained by separation and extraction from bursa of Fabricius and also can be obtained by chemical synthesis. The polypeptide has low cost and can be put into mass production. The polypeptide has an induction effect on antibody production, and also can regulate production of cytokines and proliferation of lymphocyte and promote cellular immune response. In addition, the polypeptide also regulates differentiation of B cell in a dose-dependent mode. The polypeptide has a wide application prospect, taking an application of the polypeptide in the aspects of immunomodulation, treatment and the like for example.

The invention discloses a composition based on stem cells and application thereof in preparing preparation for the coronary heart disease. The composition based on the stem cells includes the stem cells prepared through an optimizing method and cell preserving liquid serving as a solvent. Firstly, proper stem cells are selected; secondly, the selected stem cells are further processed through the optimizing method; lastly, the stem cells are collected and resuspended in the cell preserving liquid. A various cell implanting method is adopted in the application of the composition based on the stem cells in preparing the preparation for the coronary heart disease, the time nodes before an operation, during the operation and after the operation are combined, and it is ensured that the transplanted stem cells can give the full play. The optimizing method is used for preparing the stem cells and the stem cell composition in the cell preserving liquid, the ventricle function can be extremely obviously enhanced after cell transplantation, and the left ventricle reconstitution is restrained. The optimal cell composition is provided for treating the coronary heart disease, the source is wide, preparation is simple, toxic and side effects are avoided, and the transplanting effect is good.

A combined application of Haemophilus parasuis (HPS) LC strain having a deposit number of CGMCC No. 5257 and HPS LZ-20100109 strain having a deposit number of CGMCC No. 5802 in preparing a bivalent inactivated vaccine is provided, relating to HPS disease vaccines in a field of veterinary biologics. The combined application of the HPS LC strain and the HPS LZ-20100109 strain in preparing the bivalent inactivated vaccine is safe and reliable, providing not only a homologous challenge protection against serotype 1 and serotype 5, but also certain cross protection against heterologous challenges of serotype 2, serotype 4, serotype 10, serotype 12, serotype 13, serotype 14, and serotype 15 of HPS. The combined application of the HPS LC strain and the HPS LZ-20100109 strain has an obviously increased effect. After immunizing swine, a relatively strong immunity is generated; an incidence rate and a mortality rate of inoculated swine decrease obviously.

The invention discloses an active polypeptide with an antagonistic chemotactic factor receptor CCR5, which has an amino acid sequence in SEQ ID NO:1 or an amino acid sequence obtained by carrying out deletion, addition, insertion and substitution of one or more amino acid residues on the amino acid sequence in SEQ ID NO:1. The invention also discloses a preparation method of the polypeptide, which comprises the following steps of: searching related protein sequences in a database; carrying out multiple alignment comparative analysis to screen out a highly conservative region for combination of a vMIP-II receptor; eliminating interference of unrelated residues through analysis simulation means; determining a target peptide sequence; synthetizing the active polypeptide by a solid-phase synthesis method; and finally testing the bioactivity of the polypeptide. The polypeptide can be used as a CCR5 receptor antagonist for the development of an anti-inflammatory drug, can be used as a lead drug for treating AIDS (Acquired Immune Deficiency Syndrome) and can also be used as a medicament with the immune regulation effect or a medicament for improving the functions of an immune system of an organism.

The invention provides a method for preparing a rabbit acellular tracheal matrix. The method includes the following steps: using a fresh rabbit trachea as a raw material, permeating and dissolving insterile distilled water at 4 DEG C for 48 hours, entering a first decellularization cycle: soaking in 4% w/v (g/ml) of a sodium deoxycholate solution, and placing in shaking table at 20 DEG C for performing incubation for 4 h; rinsing with sterile PBS for twice, then placing in 1 mol/L NaCl containing Dnase I at a concentration of 50 kU/mL, continuously shaking for 3 h to dissolve a nucleus and degrade DNA, and finally rinsing twice with the PBS; and performing a next decellularization cycle in the same procedure, and totally performing treatment of two cycles. The method unexpectedly discovers that through improved DEM treatment, it is only required to prepare an acellular matrix similar in structure of an original trachea in the two cycles, mucosal epithelial cells are completely removed, a few chondrocytes remain, a structure is complete, the immunogenicity is weak, infiltration of inflammatory cells is less, the clinical preparation time is shortened, and a treatment basis and hopeare provided for patients who need emergency tracheal stents.

The invention relates to an adipose tissue composite preparation, and a preparation method and an application thereof. The adipose tissue composite preparation includes PDGF, umbilical cord mesenchymal stem cells, SVF and adipose tissues. The preparation method comprises the following steps: preparing a cell suspension from the SVF, the umbilical cord mesenchymal stem cells and the PDGF, and uniformly mixing the adipose tissues with the cell suspension according to a volume ratio of 20:1 in order to prepare the adipose tissue composite preparation. The adipose tissue composite preparation can be applied in plastic surgery or beauty treatment as a filling preparation. The composite biological preparation has the advantages of easiness in revascularization after being injected, and increased survival rate, is safe and reliable, allows the adipose tissues to be pale yellow, soft and bright and to be only wrapped with few fibers after being transplanted for 2-3 months; and the density of capillary tubes in the center position of transplanted fats is normal, and the blood vessel revascularization is obvious.

The invention discloses a method for inducing and differentiating precursor cells of a human skin source into corneal endothelial cells. The precursor cells of the human skin source are used and cultured together with the corneal endothelial cells B4G12, corneal endothelial cells theoretically approaching corneal endothelial cells of a normal person are successfully induced, the obtained corneal endothelial cells are applied to a corneal endothelial decompensation animal model, the corneal endothelium of an animal is successfully repaired, and the method has important clinical application prospects.

The invention discloses a recombinant rat phospholipase Cγ2 adenovirus, which is obtained by inserting the rat phospholipase Cγ2 gene into an adenovirus shuttle plasmid pHBAd‑MCMV‑GFP containing the adenovirus genome, the rat phospholipase Cγ2 gene The nucleotide sequence is shown in SEQ ID NO.1. The present invention also provides the construction method and application of the above-mentioned recombinant rat phospholipase Cγ2 adenovirus. Aiming at the promoting effect of PLCγ2 in cancer cell apoptosis, the present invention constructs a PLCγ2 recombinant adenovirus to replace the gene model, which can regulate the growth of cancer cells in a targeted manner, and the recombinant rat phospholipase Cγ2 adenovirus can stably express rat phospholipids Enzyme Cγ2, with high virus titer and significant pro-apoptotic effect; recombinant adenovirus and the rat phospholipase Cγ2 expressed by it can significantly inhibit cell proliferation and promote apoptosis of rat liver tumor cells, and can be used for the prevention of liver tumors And treatment provides new treatment ideas.

The invention provides a preparation method and application of a sclerostin single-chain antibody. The preparation method comprises the following steps: extracting anti-SOST monoclonal antibody 5H3D1hybrid tumor cell total RNA; amplifying the antibody light-chain variable region gene and heavy-chain variable region gene by an RT-PCR method; introducing Linker3 between genes VL and VH by an overlapping extending splicing PCR method; forming the single-chain antibody SOST-scFv through connection. The scFv gene with the correct sequencing is cloned to an expression vector PET-22b(+) and is thentransferred into E.coliBL21 for solubility expression; polyacrylamide gel electrophoresis SDS-PAGE is used for identifying expression products; ELISA and West-blot are used for detecting the reactionactivity of the expression protein. The preparation method and the application have the advantages that the molecular weight is small; the permeability is high; the antigen combination activity is high; the bone generation is obviously promoted; the bone loss is inhibited.

The invention belongs to the field of molecular biology, and specifically relates to an epithelial cell adhesion molecule (EpCAM) single-domain antibody and a nucleotide sequence for encoding the antibody. A phage display technology is utilized, a single-domain heavy chain antibody library capable of specifically binding to a target antigen EpCAM is screened from a single-domain heavy chain antibody library immunized with alpaca, a coding sequence of positive clone is further introduced into a prokaryotic expression vector, and therefore the high-expression EpCAM single-domain antibody is obtained. The antibody disclosed by the invention provides a basis for clinic of targeted immunological therapy of the EpCAM.

The invention relates to a method for detecting and diagnosing early-stage acute infection of toxoplasmosis by utilizing aptamers which are separately YZ-39 and YZ-56. The action mechanism of the aptamers is that the aptamers are specifically combined with a rhoptry secretory protein ROP of the toxoplasma gondii. The secretory protein ROP is the early-stage secretory protein of a toxoplasma gondii infected host cell, and is of great significance on diagnosing early-stage acute infection of the toxoplasmosis. At present, the report about detecting early-stage acute infection of toxoplasmosis by utilizing aptamers to detect the secretory protein ROP is not available at home and abroad. The discovery and application of the ROP specific aptamers provide a novel method for detecting the early-stage acute infection of the toxoplasmosis.

The invention discloses an extracting method of dog adipose-derived stem cells, and a preparation and application of the dog adipose-derived stem cells. The preparation for treating dog chronic nephrosis is prepared from the dog allogeneic adipose-derived stem cells. The extracting method of the adipose-derived stem cells comprises the steps: the dog abdominal adipose tissue is obtained, digesting, filtering and centrifuging are conducted, red blood cell lysis buffer resuspending is conducted, and the cells of P0-P3 generations are cultured; and the obtained adipose-derived stem cells of the P3 generation are subjected to surface antigen testing, the adipose-derived stem cells which have more than 70% of CD29 and MHC-1 expressions, less than 2% of CD34 and CD45 expressions, and the high differentiative potential, achieve adipogenesis, osteogenesis and chondrogenesis, and have normal chromosomes, feminine endotoxin and nont-detected mycoplasma are screened out and resuspended with the dosage of 1*10<6> cells/kg-5*10<6> cells/kg through normal saline of 0.5-1.0 mL to prepare the application of the dog allogeneic adipose-derived stem cells, and the preparation is used for treating dogchronic nephrosis. The stem cell preparation is weak in immunogenicity and is not involved in argument in the aspects of society, ethic and law; a transplanted person does not need to provide autologous stem cells, and nearly no damage is caused; and a separating method is simple, and the cells are very high in activity and easy to increase massively.