22007results about "Foreign genetic material cells" patented technology

To provide VHH-containing polypeptides that bind CD123, and a method for treating cancer that includes administration of the VHH-containing polypeptides.SOLUTION: Provided is a polypeptide comprising at least one VHH domain that binds to CD123, comprising CDR1, CDR2, and CDR3, each comprising a specific amino acid sequence.SELECTED DRAWING: Figure 3A

To provide methods for treating a subject having or at risk of developing a neurocognitive disorder, such as frontotemporal lobar degeneration or neuronal ceroid lipofuscinosis.SOLUTION: Provided is a method comprising administering, to a subject diagnosed as having a neurocognitive disorder (NCD), a composition comprising a population of cells comprising a transgene encoding a progranulin (PGRN) or a granulin (GRN).SELECTED DRAWING: Figure 1A

A recombinant humanized collagen and a preparation method therefor. The recombinant humanized type III collagen can be efficiently and stably expressed in three expression systems of yeast, Escherichia coli, and CHO cells, and has the biological activity superior to that of a commercially available collagen product. The recombinant humanized type III collagen has structural characteristics and functional activity similar to those of a natural collagen, and thus has wide application prospects in biomedical materials, tissue engineering products, cosmetics, foods, health care products or drugs, medical instruments, medical cosmetology and the like.

The invention relates to a nano antibody targeting cadherin 17 and application thereof, and relates to the technical field of immunology and molecular biology, the amino acid sequence of a complementary determining region of the nano antibody targeting cadherin 17 comprises CDR1 shown as any one of SEQ ID NO: 21-32, CDR2 shown as any one of SEQ ID NO: 34-45, and CDR3 shown as any one of SEQ ID NO: 47-58. According to the invention, an alpaca immune phage display library is constructed, a series of nano antibody molecules targeting cadherin 17 (CDH17) are screened from the alpaca immune phage display library, and functions and applications of the nano antibody are verified at various levels of CDH17 antigen protein, CDH17 target cells, CDH17 CAR-T cells and the like.

The invention provides recombinant human III-type collagen which is of a triple-helix structure and assembled into collagenous fibers and application of the recombinant human III-type collagen, and belongs to the technical field of bioengineering. Collagen fragments capable of forming a triple helix structure are screened out through a bioinformatics algorithm, the fragments are connected in series, a key C-terminal structural domain and a specific amino acid sequence are combined, and formation of a stable triple helix advanced structure and assembly of collagenous fibers are effectively promoted. The recombinant human III type collagen is efficiently expressed through a eukaryotic expression system, and shows excellent stability, low immunogenicity and good biocompatibility. Experimental results show that the prepared recombinant human III-type collagen not only has a natural triple helix structure, but also can be further assembled into macromolecular collagen fibers, is suitable for multiple fields such as medical beauty, bioengineering and tissue repair, and has wide application prospects and market values.

To provide antibodies that bind to lymphocyte activation gene 3 (LAG3) protein, which is a T cell co-inhibitor, and methods of use.SOLUTION: In various embodiments of the invention, the antibodies are fully human antibodies that specifically bind to LAG3. In some embodiments, the antibodies of the invention are useful for inhibiting or neutralizing LAG3 activity, thus providing means of treating a disease or disorder such as cancer or viral infection. The antibodies of the present invention are useful, inter alia, for targeting immune cells expressing LAG3 and for modulating LAG3 activity.SELECTED DRAWING: None

The present disclosure relates in some aspects to methods, cells, and compositions for preparing cells and compositions for genetic engineering and cell therapy. Provided in some embodiments are streamlined cell preparation methods, e.g., for isolation, processing, incubation, and genetic engineering of cells and populations of cells. Also provided are cells and compositions produced by the methods and methods of their use. The cells can include immune cells, such as T cells, and generally include a plurality of isolated T cell populations or types. In some aspects, the methods are capable of preparing of a plurality of different cell populations for adoptive therapy using fewer steps and / or resources and / or reduced handling compared with other methods.

The invention provides a lipid nanoparticle (LNP) compound for efficiently delivering nucleic acid to immune cells, and belongs to a biological medicine technology. According to the present invention, on the basis of the previously researched and developed ionizable cationic lipid 1-126, the Box-Behnken design is adopted to design 13 LNP formulas, and the microfluidic technology is adopted to complete the preparation of the LNP; by detecting the efficiency of LNP transfecting different immune cells and optimizing LNP components, the lipid nanoparticle compound for efficiently delivering mRNA in various immune cells is screened, the compound comprises 40% of cationic lipid 1-126, 16% of DSPC, 1.5% of DMG-PEG2000 and 42.5% of beta-sitosterol, and an efficient delivery system is provided for the fields of vaccine development and gene therapy.

The invention belongs to the field of molecular biology and immunology, and particularly relates to an anti-CD3 nano antibody capable of activating the activity of T cells as well as a preparation method and application of the anti-CD3 nano antibody. The invention provides a preparation method of the anti-CD3 nano antibody. The preparation method comprises the following steps: constructing a CD3 plasmid and an overexpression cell line, and immunizing alpaca to obtain peripheral blood mononuclear cells (PBMC) of the alpaca; extracting RNA (Ribonucleic Acid) and carrying out reverse transcription to obtain cDNA; carrying out PCR amplification on the antibody gene segment and cloning the antibody gene segment to a phage display vector; and electrically transforming the vector into SS320 competent cells to obtain the nano antibody library. After two rounds of panning, detecting and screening an antibody clone capable of being combined with the CD3 protein through ELISA (Enzyme-Linked Immunosorbent Assay); an antibody sequence is selected through sequencing, and an antibody protein is expressed by using a mammalian cell high-throughput expression system; and then detecting and screening the antibody which is only specifically combined with the CD3. The method disclosed by the invention is simple, convenient and rapid to operate, and can be used for preparing active antibodies for treating potential tumors and proliferative diseases.

The invention discloses a porcine muscle stem cell line capable of being cultured in vitro for a long time and application of the porcine muscle stem cell line, and belongs to the technical field of stem cells and cell culture meat. The porcine muscle stem cell line is prepared by the following steps: performing CDKN2A gene knockout on porcine muscle stem cells by using a CRISPR / Cas9 gene editing technology, performing monoclonal screening and genotype identification to obtain a cell strain with successful CDKN2A knockout, and then performing carrier-free and serum-free suspension culture domestication on the cell strain. And a carrier-free and serum-free porcine muscle stem cell line capable of being subjected to in-vitro long-term suspension culture is obtained. Verification shows that the CDKN2A knockout porcine muscle stem cell line subjected to carrier-free and serum-free suspension culture domestication not only can be subjected to carrier-free and serum-free suspension culture in a shake flask for a long time, but also can maintain a good differentiation characteristic. The porcine muscle stem cell line which can be cultured on a large scale, is derived from edible animals and can be cultured in vitro for a long time is created, and excellent cell resources are provided for commercial production of cultured meat.

The invention belongs to the field of biological medicines, and particularly relates to a varicella-zoster virus gE fusion protein as well as a preparation method and application thereof. The varicella-zoster virus gE fusion protein comprises an amino acid sequence as shown in SEQ ID NO: 1 and at least one of the following fragments: a signal peptide, a Foldon sequence, a Ferritin sequence and a His tag sequence, the signal peptide can improve the protein expression quantity, the Foldon sequence can improve the stability of the protein, the particle size of an antigen can be increased, the immunogenicity of the antigen can be improved, and the fusion protein can be used for preparing a medicine for treating the varicella-zoster virus gE fusion protein. The Ferritin sequence can enhance immune response, a vaccine prepared from the gE fusion protein has excellent immunogenicity, and compared with a commercially available vaccine Shingrix, higher humoral immunity and cellular immunity levels can be induced.

The invention provides an artificial nucleic acid molecule which is used for improving the expression quantity of target amino acid, polypeptide or protein. The artificial nucleic acid molecule at least comprises a target 5'untranslated region (UTR), a target coding region (CDS) and a target 3 'untranslated region (UTR). Wherein the sequence of the target 5 'UTR is one of the following sequences: 5' UTR of a high-expression gene and a 5 'UTR variant of the high-expression gene. The sequence of the target 3 'UTR is one of the following sequences: 3' UTR of a high-expression gene and a 3 'UTR variant of the high-expression gene. Optionally, the artificial nucleic acid molecule may further comprise, for example, a 5 '-end cap structure (Cap), a PolyA tail. The 5 'UTR and the 3' UTR have regulating effects on translation and stability of nucleic acid molecules, so that the 5 'UTR, the 3' UTR and variants thereof are selected from high-expression genes, the nucleic acid molecules can be further stabilized and are not easy to degrade, and the amount of protein or polypeptide obtained by translation of the nucleic acid molecules can be increased. The invention also provides methods for making, delivering, and using such artificial nucleic acid molecules, as well as the use of the artificial nucleic acid molecules for the treatment and / or prevention of related diseases or disorders.

The invention discloses a site for stably expressing protein in a CHO cell gene NW023276806.1 and application of the site, and belongs to the technical field of biological genes. The site belongs to a fixed position in a CHO cell genome, different protein genes are introduced based on a micro-homologous end connection mechanism through a CRISPR / Cas9 tool, and stable expression is carried out. By adopting a site-specific integration method, a target gene is integrated to a stable expression area in a site-specific manner, repeated high-expression monoclonal screening is effectively avoided, and an MMEJ mechanism is introduced to integrate a donor fragment, so that the research and development time for constructing a stable expression cell strain in biological pharmacy can be effectively shortened, and the cost is reduced.

The present invention provides a method for packaging a BaEV retroviral vector. The method includes constructing a stable BaEV retroviral packaging cell line, transfecting a retroviral vector plasmid containing a target sequence into a virus-producing cell line, where the virus-producing cell line is a HEK293T cell line or its derivative cell line, to harvest the transiently transfected and virus-producing retroviral vector, mixing it with the stable BaEV retroviral packaging cell line, and promoting the transduction of the retroviral vector into the stable BaEV retroviral packaging cell line by horizontal centrifugation. The present invention can achieve the packaging of a BAEV-type retroviral vector carrying a CD19CAR foreign gene plasmid, construct a BAEV-type retroviral vector-producing cell line, and produce a high-titer BAEV-type retroviral vector.

The invention discloses a method for improving beef quality, an action mechanism and an experimental method, in a cattle body, vitamin A can activate the expression of EBF2 through an active metabolite RA of the vitamin A, and the EBF2 can inhibit the transcription process of CYP26B1 in a targeted manner to maintain the activity of a retinol signal channel, so that PPAR gamma and downstream lipid metabolism related genes thereof are activated, and the activity of the retinol signal channel is improved. Fatty acid transport and lipid accumulation in fat cells in the cattle muscle are promoted, fat deposition in the cattle muscle is promoted, and the beef quality is improved. By constructing the molecular network for regulating and controlling the formation of the fat in the cattle muscle, a complex regulation and control mechanism for controlling the fat deposition in the cattle muscle can be understood more deeply, a method for improving the beef quality based on regulating and controlling the vitamin A mediated key factor EBF2 is provided, and a new way is provided for improving the beef quality in a targeted manner.

The invention provides a porcine epidemic diarrhea virus S protein mutant, a nucleic acid molecule, a subunit vaccine and application. Compared with wild type porcine epidemic diarrhea virus S protein, the porcine epidemic diarrhea virus S protein mutant has the advantage that amino acid at the 1076th site, amino acid at the 1077th site, amino acid at the 1016th site and amino acid at the 1017th site are mutated into proline. The natural trimer which is stable and maintains the conformation of the S protein before fusion is constructed, the safe and efficient PEDV subunit vaccine is obtained, and the use effect and immunogenicity of the porcine epidemic diarrhea vaccine can be improved; moreover, compared with a wild type S protein, the porcine epidemic diarrhea virus S protein mutant disclosed by the invention has a higher expression quantity.

The invention provides a monoclonal antibody of a p-Tau217 protein and an application of the monoclonal antibody. In particular, the present invention provides an antibody or an antigen binding fragment thereof targeting a p-Tau217 protein. The antibody provided by the invention can specifically recognize and bind the p-Tau217 protein, does not recognize p-Tau181 and non-phosphorylated Tau protein, and can be used for preparing a detection preparation or a kit for diagnosing diseases related to the p-Tau217 protein, such as Alzheimer's disease.

The invention belongs to the field of interferon gene engineering, and discloses a long-acting cat omega2 interferon recombinant protein FeIFN-omega2-Fc and application, and the FeIFN-omega2-Fc provided by the invention has the characteristics of high activity, obvious antiviral effect and long half-life period. The applicant carries out effective fusion and mutation on the basis of omega2 interferon, and finally obtains the recombinant protein FeIFN-omega2-Fc. The protein obviously activates downstream signal factors of interferon, has the effect of resisting various cat viruses, shows good safety and curative effect when cat herpes treatment experiments are carried out on cats, and compared with FeIFN-omega2, the half-life period of recombinant interferon is obviously prolonged. The FeIFN-omega2-Fc has a good antiviral effect, can effectively prevent and treat cat viral diseases, has a wide application prospect, and particularly has a relatively good popularization potential in clinical treatment.

The invention discloses a mammalian cell expression vector, an expression plasmid and an expression system. The expression vector sequentially comprises the following elements: (a) a promoter; (b) a Kozak sequence; (c) a polyadenine sequence, the polyadenine sequence sequentially comprises a bGH PolyA sequence and a 5 'UTR PolyA sequence, the nucleotide sequence of the bGH PolyA sequence is as shown in SEQ ID NO.5, and the nucleotide sequence of the 5' UTR PolyA sequence is as shown in SEQ ID NO.6; (d) copying the start site; and (e) screening marker genes. According to the present invention, the expression vector is optimized by the expression element, such that the exogenous gene can be conveniently, efficiently and flexibly inserted and edited by the expression vector so as to achieve the stable extracellular or intracellular expression of the exogenous protein, and the expression efficiency of the exogenous protein can be improved by carrying out the expression plasmid construction based on the expression vector; and more efficient expression and purification of the foreign protein are realized.

The invention belongs to the technical field of cell cryopreservation, and relates to a cryopreservation method based on ultrasonic ice planting. In the method, the nucleation temperature of ice planting is less than-9 DEG C, and the cooling rate from nucleation to icing is less than 5 DEG C / min. Compared with the prior art, the method has the advantages that the time required for cryopreservation of the cells is remarkably shortened, meanwhile, damage to the cells caused by ice crystals generated in the cooling process is reduced, and the problem that an existing cell cryopreservation cooling method cannot meet the nucleation and freezing uniformity of a cell solution in the cooling process is solved.

The invention discloses a CA9-targeted chimeric antigen receptor and application thereof, and belongs to the technical field of biological medicines. The CA9 targeting chimeric antigen receptor provided by the invention comprises a CD8 alpha leading signal peptide, an anti-CA9 single-chain variable fragment, a CD8 alpha hinge, a CD8 transmembrane structural domain and a CD3 zeta intracellular costimulatory signal transduction structural domain. According to the invention, an anti-CA9 single-chain variable fragment is used as an antigen binding structural domain to construct a chimeric antigen receptor (CAR) molecule, the macrophage expressing the anti-CA9CAR can realize specific targeting and phagocytosis killing of CA9 positive kidney cancer cells, and the constructed lipid nanoparticle delivery system can effectively transfer a gene expression vector, so that the gene expression vector can realize effective target site delivery, and the CA9 positive kidney cancer cells can be effectively targeted and killed. The constructed hydrogel-liposome combined drug delivery system has the performance of relieving toxic and side effects, controllable release and hemostasis.

The invention belongs to the technical field of biology, and provides a nanometer antibody targeting Nectin-4, and a nucleic acid molecule and an expression vector for coding the nanometer antibody targeting Nectin-4. The invention also provides a chimeric antigen receptor which comprises an extracellular antigen binding domain, a transmembrane domain and an intracellular domain, and the extracellular antigen binding domain is a Nectin-4 nano antibody; the invention also provides a chimeric antigen receptor CAR which contains the nano antibody as scFv and co-expresses CXCR1, a CAR-T cell constructed by the chimeric antigen receptor CAR, and an enhanced CAR-T cell obtained by modifying the CAR-T cell with microbial short-chain fatty acid, so as to improve the infiltration and anti-tumor activity of the CAR-T cell targeting Nectin-4.

The invention provides an anti-CDH17 antigen binding fragment, an antibody and application thereof. CDR1 of a heavy chain variable region of the antigen binding fragment comprises amino acid sequences as shown in SEQ ID NO.1-3, CDR2 of the heavy chain variable region of the antigen binding fragment comprises amino acid sequences as shown in SEQ ID NO.4-6, and CDR3 of the heavy chain variable region of the antigen binding fragment comprises amino acid sequences as shown in SEQ ID NO.7-9; cDR1 of a light chain variable region of the antigen binding fragment comprises amino acid sequences as shown in SEQ ID NO.10 to 12, CDR2 of the light chain variable region of the antigen binding fragment comprises amino acid sequences as shown in SEQ ID NO.13 to 15, and CDR3 of the light chain variable region of the antigen binding fragment comprises amino acid sequences as shown in SEQ ID NO.16 to 18. The antigen binding fragment and the anti-CDH17 antibody provided by the invention have high specificity, and a chimeric antigen receptor and a CAR-T cell prepared from the antibody have obvious cytotoxicity on cells stably expressing CDH17 protein.

The invention relates to the technical field of recombinant protein expression, in particular to recombinant human-derived III-type collagen as well as a preparation method and application thereof. The invention provides a recombinant human source III type collagen. The amino acid sequence of the recombinant human source III type collagen is shown as SEQ ID NO. 1. The recombinant human-derived III-type collagen prepared by the invention can be better applied to a human body, better exerts biological functions of the collagen, avoids repulsive interaction, is high in safety, can effectively promote cell proliferation and oxidation resistance, and is high in expression yield, stable in batches and good in quality; the recombinant human III type collagen prepared by the invention has the potential of being applied to preparation and production of biological medicines and cosmetics.

The invention discloses a nanometer antibody targeting FAP, a CAR-T cell and application of the nanometer antibody and the CAR-T cell. The invention provides nano-antibodies VHH1 and VHH2 capable of simultaneously recognizing human or mouse FAP. The nano-antibodies VHH1 and VHH2 are used as antigen binding regions to be constructed in a human or mouse CAR structure, and human or mouse FAP-CAR-T cells with a killing function can be prepared. Results of the mouse FAP-CAR-T cell or the human FAP-CAR-T cell provided by the invention show that the FAP-CAR-T cell has a good treatment effect on removal of FAP positive cells, and is a novel FAP-CAR-T cell which is expected to be applied to clinical solid tumor CAR-T therapy.

The invention relates to a humanized nano antibody targeting cadherin 17 and application of the humanized nano antibody, and relates to the technical field of immunology and molecular biology. The complementarity determining region of the humanized nano antibody comprises a CDR1 of which the amino acid sequence is shown as SEQ ID NO: 2, a CDR2 of which the amino acid sequence is shown as SEQ ID NO: 4 and a CDR3 of which the amino acid sequence is shown as SEQ ID NO: 6; a framework region of the humanized nano antibody comprises an FR1 of which the amino acid sequence is shown as any one of SEQ ID NO: 1 and SEQ ID NO: 8, an FR2 of which the amino acid sequence is shown as any one of SEQ ID NO: 3, SEQ ID NO: 9 and SEQ ID NO: 11, an FR3 of which the amino acid sequence is shown as any one of SEQ ID NO: 5 and SEQ ID NO: 10, and an FR4 of which the amino acid sequence is shown as SEQ ID NO: 7. According to the humanized nano-antibody, the immunogenicity of the nano-antibody is reduced, the targeted binding activity and killing activity are improved, and the humanized nano-antibody can be used for developing immunodetection reagents, CAR-T / NK cell drugs and antibody drugs.

The present disclosure provides inter alia, molecules comprising an autoantibody-binding domain and at least one modified Fc domain. The present disclosure also provides methods and compositions that allow for selective depletion and / or neutralization of pathogenic autoantibodies.

The invention relates to a drug delivery carrier based on extracellular vesicles and application thereof.The drug delivery carrier contains extracellular vesicle scaffold protein and recombinant membrane fusion protein, the recombinant membrane fusion protein comprises a first fragment and a second fragment, the amino acid sequence of the first fragment is shown as SEQ ID NO: 5, and the amino acid sequence of the second fragment is shown as SEQ ID NO: 6; the second fragment has n amino acids, the sequence of the second fragment is the sequence of the first n amino acids at the N end of the amino acid sequence as shown in SEQ ID NO: 7, and n is an integer between 50 and 200. According to the drug delivery carrier disclosed by the invention, the efficiency of releasing drug active molecules before reaching lysosome is remarkably improved, and the drug effect of extracellular vesicle drugs can be improved.

The invention provides a specific T cell receptor targeting KRAS G12V mutant epitope peptide (such as an amino acid sequence as shown in SEQ ID NO: 2) and anti-tumor application of the specific T cell receptor. The specific T cell receptor is composed of two peptide chains alpha and beta and can be specifically combined with a VVGAVGVGK-HLA-A * 11: 01 compound. The invention also provides an antigen binding fragment of the specific T cell receptor, a nucleic acid sequence for coding the T cell receptor, a vector containing the nucleic acid sequence, an engineered cell for expressing the T cell receptor, a composition containing the T cell receptor and application thereof. In addition, the invention also provides a method for screening the KRAS G12V specific T cell receptor. The specific T cell receptor and the antigen binding fragment thereof provided by the invention can be used as an immune effect activator to stimulate the immune response of a body, so that the effect of resisting diseases such as tumors and the like is achieved.