1137results about "Urinary tract/kidney cells" patented technology

The invention provides human placenta-derived vasogenic stem cells (hPASCs) and application thereof. The human placenta-derived angiogenic stem cells (hPASCs) disclosed by the invention are preserved in China Center for Type Culture Collection, Wuhan University, Wuhan, China; the preservation date is April 2, 2025; the preservation number of the strain is CCTCC (China Center For Type Culture Collection) NO: The hPASCs disclosed by the invention contain angiogenic cell subgroups, have stem cell stemness, have an angiogenesis effect in vivo and in vitro, and can be used for treatment of diseases such as cardiovascular diseases and vascularization construction of artificial organs and tissues.

The invention provides a UTR (Untranslated Region) sequence for improving mRNA (Messenger Ribonucleic Acid) stability and translation efficiency and application of the UTR sequence, and particularly provides an mRNA molecule which comprises a coding region for coding polypeptide or protein and a fragment thereof, a 5 'UTR sequence positioned at the upstream of the coding region and / or a 3' UTR sequence positioned at the downstream of the coding region, the 5 'UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 1, and the 3' UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 2; and / or, the 5 'UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 3, and the 3' UTR sequence is a nucleotide sequence as shown in SEQ ID NO: 4. According to the present invention, the mRNA stability and the translation efficiency are improved through the new and optimized UTR sequence, and the UTR sequence can be used as the element for enhancing the RNA expression efficiency in the nucleic acid treatment drug or the mRNA vaccine so as to provide more and better choices for the mRNA therapy.

The invention belongs to the technical field of biology, and particularly relates to a mouse hepatitis virus antibody or an antigen binding fragment thereof as well as a preparation method and application thereof. The antibody or the antigen binding fragment thereof can specifically recognize and bind to the mouse hepatitis virus or the NSP3 protein thereof, and has good affinity with the mouse hepatitis virus or the NSP3 protein thereof; the kit can be used for preparing products for diagnosing, preventing and / or treating mouse hepatitis virus infection or diseases caused by the mouse hepatitis virus infection, detecting existence or level of mouse hepatitis virus or NSP3 protein in a sample, screening medicines for preventing and / or treating the mouse hepatitis virus infection or the diseases caused by the mouse hepatitis virus infection, and carrying out basic research and / or vaccine development.

The invention discloses a bispecific nano antibody targeting CXCL1 and PD-L1 and application thereof, and belongs to the technical field of antibody engineering and bioengineering. The bispecific nano-antibody targeting CXCL1 and PD-L1 comprises at least one nano-antibody for recognizing a target CXCL1 and a nano-antibody for recognizing a target PD-L1, and nano-antibody monomers are connected through a linker. The bispecific nano-antibody targeting PD-L1 / CXCL1 provided by the invention has a unique structure, so that the bispecific nano-antibody targeting PD-L1 / CXCL1 has specific recognition and binding capacities to CXCL1 and PD-L1, the affinity to CXCL1 reaches 1.08 nM, the affinity to PD-L1 reaches 1.09 nM, drug resistance of a patient to antibody treatment caused by immune escape can be effectively prevented, and the bispecific nano-antibody targeting PD-L1 / CXCL1 is suitable for more patients and has good clinical application prospects. And a new thought is provided for CRC treatment.

The invention provides a monoclonal antibody of a targeted human folate receptor alpha (FR1). Specifically, a mouse is immunized through FR1 recombinant protein, and a monoclonal antibody with high affinity to FR1 is screened out. In addition, the invention also provides an amino acid sequence of the monoclonal antibody, nucleic acid containing the sequence, a carrier or a conjugate containing the nucleic acid, and application of the monoclonal antibody in FR1 overexpressed tumors / cancers.

The invention belongs to the field of biological medicines, and particularly discloses application of a Ptprj agonist GJ103 in preparation of a medicine for treating acute kidney injury (AKI) induced by cisplatin. In-vivo and in-vitro experiments prove that the GJ103, by activating Ptprj, can significantly down-regulate expression of pro-apoptotic protein Bax and Cleved Caspase-3, up-regulate anti-apoptotic protein Bcl2 and reduce infiltration of inflammatory factors TNF-alpha and IL-6, so that apoptosis and inflammatory response of renal tubular epithelial cells are relieved. In in-vivo experiments, GJ103 (20-40mg / kg / day) can reduce serum creatinine and urea nitrogen levels of cis-platinum model mice and improve pathological injuries such as renal tubule dilatation; in in-vitro experiments, 20-40 [mu] M of GJ103 can inhibit apoptosis of renal tubular epithelial cells and reduce expression of renal injury markers NGAL and Kim-1. The pharmaceutical composition contains GJ103 and a pharmaceutical carrier, the preparation form can be a 4mg / mL injection (the purity is greater than or equal to 99.46%) or an oral preparation, and a new strategy is provided for clinical treatment of cisplatin renal toxicity.

The invention relates to the technical field of biological medicine, in particular to an antibody aiming at staphylococcus aureus enterotoxin B and application thereof. A heavy chain variable region of the antibody provided by the invention comprises CDR sequences as shown in SEQ ID NO.1-3, and a light chain variable region of the antibody comprises CDR sequences as shown in SEQ ID NO.9-11. The antibody has high affinity, can specifically bind to staphylococcus aureus SEB, can block binding of SEB and MHC II / TCR, significantly inhibits cytokine storm, shows a dose-dependent protection effect in an MRSA systemic infection model, and can be used for preparing an MRSA systemic infection model. The monoclonal antibody can be used for treating, preventing or diagnosing the infection of the staphylococcus aureus, provides a solution of non-antibiotic therapy for SEB poisoning and drug-resistant staphylococcus aureus infection, and has important clinical and public health values.

The invention belongs to the technical field of gene engineering, and particularly relates to a biotin ligase mutant as well as a coding gene, an expression vector, a recombinant cell and application thereof. The amino acid sequence of the biotin ligase mutant is as shown in SEQ ID NO. 2. The biotin ligase (Bar A) mutant provided by the invention can be efficiently expressed in eukaryotes, the expression level of the biotin ligase (Bar A) mutant is obviously improved in mammalian cells compared with a wild type, and the biotin ligase (Bar A) mutant has good catalytic activity after expression, can effectively catalyze biotinylation of protein containing a biotin receptor sequence, and has good application prospects. The method has a wide application prospect in the fields of biotinylated protein preparation, and protein labeling, positioning, immunodetection, affinity purification, function research and the like based on a biotin-avidin system (BAS).

The invention relates to the field of immunology, and particularly discloses a TCR combination and application thereof, the TCR combination is used for recognizing HBV antigen epitopes, the TCR combination comprises a plurality of separated TCRs, the plurality of separated TCRs form two TCR groups, and the two TCR groups comprise a first TCR group and a second TCR group; wherein the first TCR group specifically recognizes a first antigen epitope, and the amino acid sequence of the first antigen epitope is as shown in SEQ ID NO: 1; the first TCR group specifically recognizes a first antigen epitope, the second TCR group specifically recognizes a second antigen epitope, the amino acid sequence of the second antigen epitope is shown as SEQ ID NO: 2, the second antigen epitope respectively and correspondingly recognizes different antigen epitopes, the hepatitis B virus specific TCR and the epitope thereof are screened out, and an important means can be provided for treatment of TCR-T immune cells infected by hepatitis B virus.

The invention provides a bovine parainfluenza virus type 3 QL3 strain, and the preservation number of the bovine parainfluenza virus type 3 QL3 strain is CGMCC No.46469. The invention also provides a bovine parainfluenza inactivated vaccine containing the bovine parainfluenza virus type 3 QL3 strain and purified F protein and HN protein of the QL3 strain. The novel bovine parainfluenza virus type 3 inactivated vaccine is obtained by optimizing the antigen components of the bovine parainfluenza virus type 3 inactivated vaccine, and high-level neutralizing antibodies can be generated after immunizing cattle; the safety of the vaccine is improved through process optimization. Safety and potency test results show that after the inactivated vaccine is used for immunizing cattle, no adverse reaction exists, and high-level neutralizing antibodies are generated. Results show that the immune effect of the bovine parainfluenza virus type 3 inactivated vaccine can be improved.

The present invention relates to antibodies and antibody-drug conjugates, and more particularly to antibodies and antibody-drug conjugates (ADCs) targeting TROP2 and / or NECTIN4 as well as compositions containing said antibodies or ADCs and therapeutic applications thereof.

The invention relates to the technical field of antibodies, and provides an anti-CD45 antibody and application thereof, the antibody comprises at least one of the following CDRs: a heavy chain variable region CDR: an amino acid sequence of SEQ ID NO: 1-3 or a conservatively modified form thereof; and a light chain variable region CDR: SEQ ID NO: 4-6 or an amino acid sequence in a conservative modification form thereof. The antibody or the antigen binding fragment thereof can efficiently and specifically recognize CD45, can effectively detect CD45, and can be used for immunohistochemistry exemplarily.

The invention relates to the technical field of cell engineering, and particularly discloses a construction method and application of a sika deer immortalized renal epithelial cell line, and the construction method comprises the following steps: taking renal epithelial primary cells from healthy sika deer renal cortex tissues to obtain primary renal epithelial cells; treating the primary renal epithelial cells with trypsin to obtain a cell suspension; inoculating the cell suspension into a complete culture medium, and transfecting with lentivirus loaded with SV40 large T antigen genes to obtain transfected cells; 3 [mu] g / mL puromycin is applied to the transfected cells for selective culture, so that non-transfected cells are eliminated, and drug-resistant cells are obtained; carrying out continuous passage on the drug-resistant cells for at least 30 generations to obtain the immortalized renal epithelial cell line of the sika deer; according to the invention, through a specific action mechanism (inhibiting a p53 / pRb pathway and blocking cell cycle exit) of the SV40 large T antigen, a proliferation limit caused by inhibition of telomerase activity of primary cells of the cervidae animals is overcome, and a cell resource library capable of realizing continuous passage is established.

The invention belongs to the technical field of biological medicine, and particularly relates to an engineered cell microvesicle and a delivery system based on the engineered cell microvesicle, the system realizes efficient preparation of 1-5 [mu] m cell microvesicles, and the cell microvesicles have a large space volume and can be used for preparing the cell microvesicles. The carrier can be used for loading and delivery of target protein, polypeptide and recombinase which are specifically expressed in mother cells. The system transfects mother cells through lentivirus transfection or plasmid transfection to further produce cell microvesicles, and the microvesicles can load more goods and inherit membrane proteins of the mother cells, and can also effectively load intracellular proteins to realize effective delivery. By virtue of good structural stability, high immunogenicity and excellent biocompatibility, the cell microvesicle reduces systematic toxic and side effects of a traditional carrier, is expected to become an effective drug delivery system, and has great application potential in the field of gene therapy.

The invention provides brain cell specific targeting delivery based on exosome, and the exosome secreted by 293T cells is used for engineering transformation, so that the capabilities of penetrating BBB and specifically targeting neurons are achieved. According to the present invention, the L1CAM is loaded on the exosome, the exosome is obtained by transfecting a 293T cell of a Lamp2b-L1CAM PCDH-HygroHA plasmid through a 293T-Lamp2b-L1CAM cell line, and the BBB cell-penetrating peptide 4F-T7 and the neuronal targeting protein L1CAM are loaded on the surface of the exosome so as to treat the neurodegenerative diseases such as the Alzheimer's disease and the Parkinson's disease.

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

The invention relates to a Vero cell serum-free culture medium and application thereof, the serum-free culture medium is suitable for free suspension growth of Vero cells and is free of animal sources and protein components, adherent cells can be directly domesticated through a certain method to screen out Vero suspension cells, and serum-free suspension culture of the Vero suspension cells is achieved in the serum-free culture medium. The density of cells subjected to suspension culture by adopting the serum-free culture medium can reach more than 107 cells / mL, the cell viability is more than 90%, the cells are well dispersed, the cells can still normally grow and proliferate after continuous subculture for 20 generations, and the cells are sensitive to various viruses.

The present invention relates to methods of preparing a therapeutic exosome using a protein newly-identified to be enriched on the surface of exosomes. Specifically, the present invention provides methods of using the proteins for affinity purification of exosomes. It also provides methods of localizing a therapeutic peptide on exosomes, and targeting exosomes to a specific organ, tissue or cell by using the proteins. The methods involve generation of surface-engineered exosomes that include one or more of the exosome proteins at higher density, or a variant or a fragment of the exosome protein.

The invention relates to the technical field of cell culture, in particular to a dynamic culture device and method for organoids, the dynamic culture device comprises a culture bin and a transverse flow reaction generator, the end, away from the transverse flow reaction generator, of the culture bin is communicated with a liquid supply bin, and the two ends of the culture bin are connected with a second driving unit through a transmission mechanism; the transverse flow reaction generator is rotationally connected with a first driving unit; and vortex bulges / grooves are formed in the inner wall of the culture bin. Under the driving of the second driving unit, the culture bin is driven to rotate; the transverse flow reaction generator is driven by the first driving unit to reciprocate in the axis direction of the culture bin, and liquid in the culture bin collides with the vortex protrusions / grooves in the inner wall of the culture bin under the transverse effect of the transverse flow reaction generator, so that transverse vortexes are formed. The device is simple in structure, can create a culture environment for providing specific mechanical stimulation for the organoid, is beneficial to amplification of the organoid, and improves the structure and function maturity of the organoid.

The invention discloses a prebiotic-loaded intestinal targeting exosome and a preparation method and application thereof.The preparation method comprises the steps that firstly, plasmids containing intestinal cell targeting peptide sequences are constructed through a gene editing technology, the plasmids are transfected and introduced into cells, the transfected cells are screened with antibiotics, and cell strains stably expressing intestinal cell targeting peptides are obtained; then culturing and adding prebiotics, and continuously culturing to enable the cells to take in the prebiotics and release the exosomes loaded with the prebiotics; and finally, collecting cell culture supernate, and centrifuging and resuspending for multiple times under a low-temperature condition to obtain the prebiotics-loaded intestinal targeting exosome. According to the preparation method, extraction of the exosome, loading of the prebiotics and an intestinal targeting modification process are optimized, so that the prepared intestinal targeting exosome loaded with the prebiotics can be used for remarkably improving the loading efficiency and intestinal targeting of the prebiotics and enhancing the stability of the exosome in gastrointestinal tracts; therefore, the delivery efficiency and the treatment effect of the prebiotics in the intestinal tract are improved.

The invention relates to an engineered exosome loaded with specific cytokines as well as a preparation method and application of the engineered exosome, and belongs to a biopharmaceutical technology. The engineered exosome is prepared by the following steps: (1) constructing an expression vector or lentivirus carrying an artificially optimized coding gene of a hepatocyte growth factor; (2) transfecting host cells, and screening successfully transfected host cells; (3) carrying out amplification culture on the selected transfected host cells, and separating and collecting exosomes of the transfected host cells; the expression vector is a transient expression vector or a lentiviral vector; an ExoBoost cell is adopted as the host cell. Experimental data show that the engineered exosome provided by the invention can promote hair follicle growth and has the application potential of improving AGA symptoms.

The invention relates to the technical field of preparation of organoids, and discloses a method for regulating and controlling human induced pluripotent stem cells to differentiate into kidney organoids, which comprises the following steps: resuspending a cell cluster formed by human induced pluripotent stem cells by using a basic differentiation culture solution containing a GSK-3beta inhibitor, inoculating the cell cluster into an ultralow adsorption cell culture plate, and starting to culture, marking the time as the 0th hour of differentiation; and after culturing for 70-76 hours, replacing the basic differentiation culture solution containing the GSK-3beta inhibitor with a basic differentiation culture solution containing a Knockout TM serum substitute (KSR), and continuing to carry out differentiation culture. By adopting the method disclosed by the invention, the human induced pluripotent stem cells can be efficiently, quickly, simply, conveniently and stably subjected to directional induced differentiation into kidney organs containing podocytes, renal tubules and other structures, and the time for constructing a kidney in-vitro model can be shortened, so that the research on kidney diseases, kidney development and the like can be greatly assisted; and a powerful platform is provided for kidney toxicity assessment.

The invention relates to the field of biomedicine, and discloses a method for screening KRAS mutant interacting proteins, an HEK293T cell line for continuously expressing an HA-Turbo ID-KRAS variant is established by utilizing lentivirus transduction, the stability of transgene expression is ensured through puromycin screening, and variation caused by transient transfection is reduced as much as possible. After a Turbo ID mediated proximity marker is activated by biotin treatment, KRAS proximity binding proteins are covalently biotinylated, captured by NeutrAvidin bead purification, and analyzed by quantitative mass spectrometry. According to the invention, two potential KRAS mutation treatment target proteins of LZTR1 and LAMTOR1 can simultaneously inhibit activation and efficacy exertion of three different mutants of G12C, G12D and G12V of the G12 mutant, compared with drugs on the market that a single target of G12C directly targets the KRAS mutant, a brand new KRAS mutation targeting intervention strategy is provided, the clinical curative effect can be further improved, and the occurrence rate of secondary drug resistance can be effectively reduced.

The invention discloses a human normal kidney immortalized cell line NRC-X1 and application thereof, and belongs to the field of microbial animal cell lines. The human normal kidney immortalized cell line is named as NRC-X1, and the preservation number is CCTCC (China Center For Type Culture Collection) NO: C2025168. The invention also discloses the application of the human normal kidney immortalized cell line NRC-X1 in a virus infection mechanism. The invention also discloses application of the human normal kidney immortalized cell line NRC-X1 in research of drug renal toxicity evaluation. The invention also discloses the application of the human normal kidney immortalized cell line NRC-X1 in bioartificial kidney and cell therapy. The invention also discloses application of the human normal kidney immortalized cell line NRC-X1 in research of kidney physiological and pathological mechanisms.

In certain embodiments, a lentiviral vector for the treatment of sickle cell disease (SCD) is provided. In certain embodiments, the vector comprises an expression cassette that encodes that an anti-sickling β-globin gene and an shRNA that inhibits expression of a BCL11A gene (BCL11A shRNA) wherein said expression cassette is in reverse orientation in the vector; a β-globin locus control region (LCR) comprising a reduced length hypersensitive site 1 (HS1) sequence, a reduced length hypersensitive site 2 (HS2) sequence, a reduced length hypersensitive site 3 (HS3) sequence, and a reduced length hypersensitive site 4 (HS4) sequence, where said anti-sickling β-globin gene is operably linked to the human β-globin locus control region.

The invention relates to bispecific antibodies comprising a first antigen-binding site that specifically binds to PD1 and a second antigen-binding site that specifically binds to TIM3, in particular to bispecific antibodies, wherein the bispecific antibody binds to 5 TIM3 with a lower binding affinity when compared to the binding to PD1. The invention further relates to methods of producing these molecules and to methods of using the same.

The invention discloses a recombinant protein preparation for optimizing a human pluripotent stem cell-derived kidney organoid, which comprises ANXA2, a human pluripotent stem cell-derived kidney organoid and a human pluripotent stem cell-derived kidney organoid, it is found for the first time that ANXA2 can significantly improve fate specialization of early differentiated cells to kidney lineages, significantly improve the stability of kidney organoid products obtained in different experiment batches, and significantly improve the overall vascularization level and maturity degree of kidney organoid products in the final differentiated stage; a new thought and strategy are provided for construction of the human pluripotent stem cell-derived kidney organoid, and the human pluripotent stem cell-derived kidney organoid has a wide application prospect.

The invention discloses a reagent for reducing the agglomeration rate of suspended HEK293F cells, a culture method and a culture medium. The reagent comprises the following components: 0.01-0.1% (v / v) of a nonionic surfactant, 0.05-0.2% (v / v) of polysaccharide or a derivative thereof, 1-5mM of L-glutamine or a derivative thereof, 0.001-0.01 mM of FeCl3, 14-16mM of HEPES and 50-100mu M of an RGD peptide inhibitor. According to the anti-caking agent, the nonionic surfactant, the polysaccharide or the polysaccharide derivative, the L-glutamine or the L-glutamine derivative, the FeCl3, the HEPES and the RGD peptide inhibitor are optimally proportioned, so that the HEK293F cell caking rate is remarkably reduced to 30% or below, after the anti-caking agent is added, cells are in a uniform single-cell suspension state, macroscopic cell agglomerates do not exist, and the anti-caking effect is good. The uptake efficiency of cells on nutrient substances is obviously improved. Compared with a commercially available PEG (Polyethylene Glycol)-containing anti-caking agent, the anti-caking agent disclosed by the invention has the advantages that the anti-caking rate is lower than 35% (the anti-caking rate is only 30% vs after being cultured for 48 hours, and the anti-caking rate is 40%) while the cell viability is maintained to be 95% or above, and no cytotoxic accumulation exists after long-term passage (more than or equal to 20 generations).

The invention discloses a fusion protein taking peptide-N-glycosidase as an active component as well as a preparation method and application of the fusion protein, and belongs to the technical field of biological medicines. The fusion protein comprises: (a) peptide-N-glycosidase or a catalytically active fragment thereof; (b) an immunoglobulin Fc domain, or a combination of a tumor or immune cell antigen binding domain and an immunoglobulin Fc domain; (c) a linker peptide; wherein the form of the tumor or immune cell antigen binding domain is Fab, scFv or VHH; the peptide-N-glycosidase or the catalytic activity fragment of the peptide-N-glycosidase is connected with the Fc structural domain of the immunoglobulin through the connecting peptide; the immunoglobulin Fc domain mediates the fusion protein to form a homodimer or a heterodimer. According to the invention, the synergistic function of targeted binding and local deglycosylation of the target molecule is realized, so that the immunosuppressive activity of the target molecule is interfered, and the anti-tumor immune response is enhanced.

The invention belongs to the technical field of biological medicine, and particularly relates to an in-vitro fusion technology for multi-lineage human kidney organs and application thereof. The invention provides a method for realizing culture of multilineage kidney organs by fusing kidney units and ureteral bud organs in vitro. According to the in-vitro fusion technology of the kidney organoid reported by the invention, two organoid with different pedigree sources, namely the kidney unit and the ureteral bud, can be efficiently fused, a UB tubular structure is integrated into the kidney unit organoid, and the UB tubular structure starts from a single ureteral bud and is wrapped by kidney unit precursor cells in a similar in-vivo development process, and finally develops into a kidney with glomerulus and a plurality of kidney cells. The multi-lineage renal organ comprises a catheter, a proximal tube, a marrow tab thin section, a distal tube, a connecting tube and a ureteral bud with a single outlet. A framework is provided for an engineered renal unit and a smooth collection system, and the method is an important step for generating the functional renal tissue from the beginning.