700 results about "Mammalian cell" patented technology

The invention relates to the technical field of biological medicines, and particularly discloses an anti-human CD3 rabbit recombinant monoclonal antibody as well as a preparation method and application thereof. The antibody obtains anti-CD3 immune response by immunizing a rabbit, is obtained by combining single B cell sorting, gene cloning and a mammalian cell expression system, and has high affinity and good specificity. The antibody can effectively recognize CD3 positive T cells in human peripheral blood and can be applied to the fields of flow cytometry, immunohistochemistry, T cell function research and the like. Experimental results show that the antibody has the advantages of high titer, strong fluorescence signal, low background and the like, and is superior to the traditional murine anti-CD3 antibody. Amino acid sequences of light chain and heavy chain variable regions of the antibody are further obtained, and controllability and repeatability of antibody expression are achieved. The antibody has important scientific research value and industrial application prospect, and is suitable for various scenes such as T cell marker detection, immune state evaluation, immunotherapy auxiliary research and the like.

The present invention belongs to the technical field of bioengineering. Provided are an IscB mutein and the use thereof. Compared to the wild-type IscB protein, the IscB mutein comprises arginine at at least one of the following amino acid positions: position 84, position 96, position 102, position 111, position 159, position 368 and position 386; preferably, the IscB mutein further comprises arginine at position 401 and position 456. The relatively strong interaction between the IscB mutein and a double-stranded DNA target improves the editing activity for mammalian cell lines, which can greatly enhance the application potential of the IscB mutein as a new-generation underlying tool for gene editing, and promote the use of the IscB mutein in the aspects of precision medicine, animal model construction and crop breeding.

The invention relates to a triple-hole paper-based three-dimensional cell culture chip as well as a preparation method and application thereof. The chip comprises a paraffin hydrophobic region and a paper-based hydrophilic region. The preparation method mainly comprises the following steps: 1) designing a mask plate; 2) cutting the mask plate by laser; and 3) preparing the triple-hole paper-based chip by hot pressing. The thickness of the chip is 100-200 [mu] m, the porous fiber structure of the paper-based material allows transportation of oxygen and waste, and a three-dimensional microenvironment for in-vivo cell growth can be simulated. Meanwhile, a transparent and liquid-tight supporting layer is fused on the lower layer of the paper-based chip, so that leakage of cells inoculated on the upper layer of the paper-based chip is avoided. The paper-based chip is embedded into a 6-pore plate, and bladder cancer cells (5637), mouse astrocytoma (U87) and human breast cancer cells (MCF-7) are inoculated in a paper-based hydrophilic region respectively. The cell compatibility of the paper-based chip and the growth behaviors of three different types of cells in three-dimensional paper fibers are investigated. It is proved that the chip can be used for researching three-dimensional construction and growth of human and mammalian cells, and the difference between three-dimensional culture and two-dimensional culture is revealed by detecting the proliferation capacity and survival rate of different cells.

Reported herein is, inter alia, a method for producing a trivalent bispecific antibody, the method comprising the steps of culturing a mammalian cell comprising a deoxyribonucleic acid encoding the trivalent bispecific antibody, and recovering the trivalent bispecific antibody from the cell or culture medium, wherein the deoxyribonucleic acid encoding the trivalent bispecific antibody is stably integrated into the genome of the mammalian cell and comprises in the 5' to 3' direction a first expression cassette encoding a first heavy chain, a second expression cassette encoding the first heavy chain, a third expression cassette encoding a first light chain, a fourth expression cassette encoding the first light chain, a fifth expression cassette encoding the second heavy chain, a sixth expression cassette encoding the first light chain or the second heavy chain or the second light chain, and a seventh expression cassette encoding the second light chain, wherein the first heavy chain The chain comprises, from N-terminus to C-terminus, a first heavy chain variable domain, a CH1 domain, a hinge region, a CH2 domain, a CH3 domain, a peptide linker, a second heavy chain variable domain and a CL domain, the second heavy chain comprises, from N-terminus to C-terminus, a first heavy chain variable domain, a CH1 domain, a hinge region, a CH2 domain and a CH3 domain, the first light chain comprises, from N-terminus to C-terminus, a first light chain variable domain and a CH1 domain, and the second light chain comprises, from N-terminus to C-terminus, a second light chain variable domain and a CL domain, wherein the second heavy chain variable domain and the first light chain variable domain form a first binding site, and the first heavy chain variable domain and the second light chain variable domain form a second binding site.

The invention relates to the technical fields of gene engineering, antibody engineering and immunodiagnosis. The invention provides a recombinant humanized anti-Cpn IgM monoclonal antibody. The monoclonal antibody comprises a light chain variable region and a heavy chain variable region, the light chain variable region comprises three complementary determining regions CDR1, CDR2 and CDR3, and the amino acid sequences of the three complementary determining regions are respectively shown as SEQ ID NO.3, SEQ ID NO.4 and SEQ ID NO.5; the heavy chain variable region comprises three complementary determining regions CDR1, CDR2 and CDR3, and the amino acid sequences of the three complementary determining regions are shown as SEQ ID NO.6, SEQ ID NO.7 and SEQ ID NO.8 respectively. The invention also provides a method for preparing the monoclonal antibody. The recombinant humanized anti-Cpn IgM monoclonal antibody provided by the invention is expressed in mammalian cells by utilizing a gene recombination technology, has the characteristics of high purity, small batch difference and the like, and can be applied to a Cpn IgM antibody detection kit as a quality control product; as a quality control product, the recombinant humanized anti-Cpn IgM monoclonal antibody has the advantages of high sensitivity, high safety, inter-batch stability and the like.

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 discloses a decafluorobiphenyl-based Tat47-57 binding peptide, the amino acid sequence of the binding peptide is shown as SEQ ID NO.1 or SEQ ID NO.2, the fourth amino acid and the eighth amino acid of the SEQ ID NO.1 are bound through decafluorobiphenyl, and the fifth amino acid and the twelfth amino acid of the SEQ ID NO.2 are bound through decafluorobiphenyl. In particular, the decafluorobiphenyl-based Tat47-57 binding peptides (P3 and P6) are efficient CPPs, capable of delivering a variety of cargo molecules into the cytoplasm of mammalian cells. Meanwhile, decafluorobiphenyl cyclization plays a key role in cellular uptake and endosome escape. The two peptides P3 and P6 show remarkable cell permeability, and are speculated to enter cells through gridding protein dependent endocytosis. They also show efficient endosome escape ability, and have successfully applied P3 and P6 as CPPs to deliver phospholipid peptides and avidin.

The disclosure features novel methods of producing nucleic acid lipid nanoparticle (LNP) compositions employing a modifying agent after formation of a precursor nucleic acid lipid nanoparticle, the produced compositions thereof, and methods involving the nucleic acid lipid nanoparticles useful in the delivery of therapeutics and / or prophylactics, such as a nucleic acid, to mammalian cells or organs to, for example, to regulate polypeptide, protein, or gene expression.

The invention relates to the technical field of gene editing, in particular to a Cas9 protein mutant and application thereof. According to the present invention, the mutation is introduced at the position far away from the catalytic site to induce the Cas9 protein conformation change so as to obtain the new Cas9 protein mutant, only the Cas9-sgRNA target chain is cut, and the Cas9-sgRNA non-target chain is not cut, such that the new Cas9 protein mutant can be used for constructing the novel base editor, and the high-efficiency base editing can be performed on the mammalian cell genome. The Cas9 protein mutant disclosed by the invention is fused with a mutant TX with DNA binding deletion of 3 '-> 5' exonuclease TREX2, so that the gene editing efficiency of a 3'terminal generated by paired single-nick enzyme cutting can be greatly improved, and the safety of the Cas9 protein mutant is maintained.

The invention provides a method for obtaining engineered recombinant mammalian cells and application of the engineered recombinant mammalian cells, and relates to the technical field of biology. The method comprises introducing into a mammalian cell a minicircle DNA comprising a transposon and a polynucleotide encoding a transposase, the transposon comprising a fragment encoding an antigen binding domain. The method has the advantages of shortening the preparation period of recombinant mammalian cells, simplifying the process and reducing the cost.

Disclosed are capsid-modified rAAV expression vectors, as well as infectious virions, compositions, and pharmaceutical formulations containing them. Also provided are methods of preparing and using the disclosed capsid-protein-mutated rAAV constructs in a variety of diagnostic and therapeutic modalities, including, inter alia, as mammalian cell-targeting delivery agents, and as human gene therapy vectors. Also disclosed are large-scale production methods for capsid-modified rAAV expression vectors, viral particles, and infectious virions having improved transduction efficiencies over those of the corresponding, un-modified, rAAV vectors, as well as use of the disclosed compositions in the manufacture of medicaments for a variety of in vitro and / or in vivo applications.

The disclosure features novel lipids and compositions involving the same. Nanoparticle compositions include a novel lipid as well as additional lipids such as phospholipids, structural lipids, and PEG lipids. Nanoparticle compositions further including therapeutic and / or prophylactics such as RNA are useful in the delivery of therapeutic and / or prophylactics to mammalian cells or organs to, for example, regulate polypeptide, protein, or gene expression.

The utility model discloses a bioreactor bubble-free air interchanger which comprises a shell, an air inlet pipe extends into the shell, an air outlet port of the air inlet pipe extends into the shell, the top of the shell is connected with an exhaust pipe, an air inlet port of the exhaust pipe is located at the top of the shell, a liquid passing hole is formed in the cylinder wall of the shell, and the liquid passing hole is communicated with the air inlet port of the air inlet pipe. A filtering body for liquid to pass through is arranged on the periphery of the shell and is of a porous net structure. Compared with the prior art, the bubble-free air interchanger of the bioreactor has the advantages that the bubble-free air interchanger of the bioreactor is made of 316 stainless steel materials, is small in occupied space and easy to install and operate, can achieve bubble-free air interchange in the bioreactor, eliminates bubble damage to cells while meeting the requirement of a culture system for oxygen concentration, and improves the air interchange efficiency of the bioreactor. The device is suitable for the culture process of mammalian cells sensitive to shear force. The device is flexible in application, can be adapted to bioreactors of different specifications, and is easy for process development and amplification.

The invention relates to the field of preparation of materials for articular cartilage repair, in particular to collagen and chitosan cross-linked temperature-sensitive gel for articular cartilage repair and a preparation method of the collagen and chitosan cross-linked temperature-sensitive gel. The cartilage repair material prepared by the invention adopts the recombinant human collagen with a triple helix structure as a main raw material. The raw material is prepared by using a mammalian cell expression system, is close to protein folding and polymerization of natural protein, and has a spatial structure and modification necessary for active protein. The recombinant human collagen with the triple-helix structure is good in biocompatibility and free of rejection reaction and anaphylactic reaction, and the risk of disease transmission possibly caused by animal-derived collagen can be avoided. The injection type temperature-sensitive gel is liquid at low temperature and is converted into hydrogel at 37 DEG C, so that the injection type temperature-sensitive gel is suitable for being used as a transfer carrier for in-vivo injection treatment, secondary injury of an organism is effectively avoided, and the injection type temperature-sensitive gel has a wide clinical application prospect.

To provide a shellac modified object which can be changed from a state where cell adhesiveness to a mammalian cell is high, to a state where the cell adhesiveness is low.SOLUTION: A shellac modified object is expressed by a formula (1). In the formula, Z is a part excluding a carboxy group modified by a functional group Y in the structure of the shellac, X1 to X4 are a dialkylamino group, alkylamino group, hydroxy group, halogen group, H or the like.SELECTED DRAWING: Figure 1

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

The present disclosure provides a composition containing a purified population of plant-derived exosome-like nanoparticles isolated from tissue of a vascular plant, wherein the exosome-like nanoparticles comprise a tuned cargo comprising a protein signature and an miRNA signature, wherein the tuned cargo of the plant-derived exosome-like nanoparticles is a result of exposure of the plant to combinations of abiotic stress conditions that cause the plant to modulate its signaling pathways and metabolism to ensure its survival in a challenging environment. The tuned cargo of the plant-derived exosome-like nanoparticles can modulate bioactivities of mammalian cells directly or indirectly. The present disclosure also provides a method for improving appearance of human skin and human hair health, including eyelashes and eyebrows, comprising applying a composition comprising the abiotically stressed plant-derived exosome-like nanoparticles containing the tuned cargo and a carrier; and applying the composition topically.

Platelet lysate compositions and cell culture media compositions for maintaining and / or growing mammalian cells, such as mammalian endothelial cells (ECs) and mammalian endothelial progenitor cells (EPCs), in particular human ECs (huECs) and human EPCs (huEPCs), such as primary huECs and primary huEPCs, are provided. The cell culture media compositions contain a basal medium, a platelet lysate and, optionally, one or more exogenously added growth factors. Also provided are methods for making and using such cell culture media compositions to grow and / or maintain ECs and EPCs, including huECs and huEPCs, as well as cell culture vessels, dishes, plates, and / or flasks pretreated with the cell culture media compositions.

Provided are a fusion protein comprising recombinant porcine interferon λ1 and a use thereof, and a preparation method and use for the fusion protein. The fusion protein is formed by respectively linking porcine interferon λ1 to other fragments (such as porcine interferon γ) and / or porcine serum albumin (PSA) directly or indirectly by means of linker elements. The porcine interferon λ1 fusion protein can be prepared by using a mammalian cell expression system on the basis of genetic engineering technology. Compared with natural porcine interferon λ1, the provided recombinant porcine interferon λ1 fusion protein has a long half-life period and can be used for preparing drugs for preventing or treating porcine viral diseases.

The invention discloses a multi-functional domain fusion extracellular matrix protein as well as a coding gene, a preparation method and application thereof, and belongs to the fields of biotechnology, medical cosmetology and wound repair. The multi-functional-domain fusion extracellular matrix protein provided by the invention contains core functional domains of human-derived III-type collagen, elastin and fibronectin, integrates key biological functions of the human III-type collagen, elastin and fibronectin, and is obtained by expression of mammalian cells. The method can promote formation of a collagen triple helix structure, is simple in process, high in yield and easy for industrial production, and can be widely applied to the fields of effective skin care, skin aging resistance, medical cosmetology, wound repair and the like.

The present disclosure relates to a mammalian cell which is modified to express on the surface of its membrane a binding domain which binds to a target molecule. The disclosure also relates to protein constructs and nucleic acids for producing such modified mammalian cells, and to methods for using the mammalian cells to deliver therapeutic agents to target cells or tissues in vivo.

The present invention relates to a method for producing higher titers and / or a higher quality of recombinant viruses and / or less or reduced impurities in virus like particle (VLP) production in respective production processes in mammalian cell lines, the method comprising the step of supplementing a suitable mammalian cell line culture media with at least one intermediate substrate of the tricarboxylic acid (TCA) cycle and optionally supplementing the suitable mammalian cell line culture media with further additives and / or further adjusting the conditions of the mammalian cell line culture media. Furthermore, the present invention relates to the balanced suitable mammalian cell line culture media and the use of this balanced suitable mammalian cell line culture media for producing higher titers and / or a higher quality of recombinant viruses and / or less or reduced impurities in virus like particle (VLP) production in respective production processes for producing higher titers and / or quality of recombinant viruses.

This document relates to materials and methods for controlling ligand gated ion channel (LGIC) activity. For example, modified LGICs including at least one LGIC subunit having a modified ligand binding domain (LBD) and / or a modified ion pore domain (IPD) are provided. Also provided are exogenous LGIC ligands that can bind to and activate the modified LGIC, as well as methods of modulating ion transport across the membrane of a cell of a mammal, methods of modulating the excitability of a cell in a mammal, and methods of treating a mammal having a channelopathy.

The invention provides an anti-influenza B virus nano antibody and application thereof, the nano antibody comprises a heavy chain variable region, the heavy chain variable region comprises CDR1, CDR2 and CDR3, and the CDR3 of the heavy chain variable region is selected from an amino acid sequence as shown in SEQ ID NO: 8 or SEQ ID NO: 16. The nano antibody is obtained through a multi-antigen cross immunization and panning method, the nano antibody can be specifically combined with Colorado / 06 / 2017 and Florida / 4 / 2006 influenza B viruses, the sensitivity is high, the specificity is high, and the phenomenon of missing detection in clinical detection can be avoided in clinical application. The nano antibody can be efficiently expressed through prokaryotic cells, and compared with eukaryotic mammalian cell expression of a traditional antibody, the nano antibody is lower in production cost and more beneficial to popularization and application in detection of infectious diseases.

The present inventions provide eukaryotic cells, such as mammalian cells, that comprise adeno-associated virus (AAV) polynucleotides, including AAV capsid proteins (Cap), and are capable of expressing the polypeptides encoded by the AAV polynucleotides, and thereby are capable of producing AAV, including recombinant AAV. The eukaryotic cells also may comprise adenovirus (Ad) polynucleotides. The present inventions also provide methods of expressing AAV polynucleotides, as well as Ad polynucleotides, in eukaryotic cells, such as CHO cells, HEK 293 and BHK cells. The present inventions further provides other products and methods described herein.

This application relates to a method for producing a protein containing an unnatural amino acid (UAA), the method comprising culturing a host cell, wherein the host cell is a eukaryotic cell, together with: a nucleotide sequence encoding a first recoding tRNA or a first recoding tRNA, wherein the first recoding tRNA comprises an anticodon complementary to a first codon, and wherein the first codon is a rare codon; and a nucleotide sequence encoding a first aminoacyl-tRNA synthetase or a first aminoacyl-tRNA synthetase, wherein the first aminoacyl-tRNA synthetase is capable of charging the first recoding tRNA with the first unnatural amino acid.

This document provides methods and materials for manipulating telomere length. For example, this document provides methods and materials for using one or more agents that can manipulate (e.g., increase or decrease) telomere length. In some cases, one or more agents that can increase telomere length can be contacted with a cell to increase telomere length in that cell. For example, one or more agents that can increase telomere length can be administered to a mammal having one or more TBDs to increase telomere length in cells within the mammal (e.g., to treat the mammal). In some cases, one or more agents that can decrease telomere length can be contacted with a cell to decrease telomere length in that cell. For example, one or more agents that can decrease telomere length can be administered to a mammal having a disease or disorder associated with increased telomere synthesis and / or increased telomerase activity to decrease telomere length in cells within the mammal (e.g., to treat the mammal).

The invention belongs to the technical field of antibody preparation, and particularly relates to an anti-CD38 nano antibody as well as a preparation method and application thereof. The method comprises the following steps: constructing and expressing CD38 antibody protein, immunizing alpaca, obtaining immune alpaca PBMC (peripheral blood mononuclear cells), extracting RNA (ribonucleic acid) of the cells, carrying out reverse transcription to obtain cDNA (complementary deoxyribonucleic acid), carrying out PCR (polymerase chain reaction) to obtain antibody gene segments, constructing the antibody gene segments on a phage expression vector to construct a CD38 immune antibody library, carrying out two rounds of panning, carrying out Clone ELISA (enzyme-linked immuno sorbent assay) detection on clones to obtain an antibody combined with the CD38 protein, carrying out sequencing analysis, and selecting a unique sequence. High-throughput expression is carried out through a mammalian cell high-throughput expression system, antibody protein is obtained, FACS detection is carried out through a CD38 high-expression cell line, and finally the antibody combined with CD38 is obtained. The method is simple and rapid to operate, a mammalian cell expression system induces efficient expression of the antibody, processing modification can be performed after translation, and the activity of the antibody is closer to that of a natural antibody.

Provided herein are engineered Epstein-Barr virus nuclear antigen 1 (EBNA1), coding molecules thereof, vectors and mammalian cell expression systems comprising the same, and polypeptide of interest recombinantly produced by the foregoing. Also provided are methods for the preparation of the engineered EBNAls, coding molecules thereof, vectors and mammalian cell expression systems and methods for using the same in recombinant expression.

To provide a recombinant polynucleotide for treating a particular disease of ectopic tissue calcification, and to provide a viral vector containing the recombinant polynucleotide.SOLUTION: Provided is a recombinant polynucleotide encoding a precursor polypeptide comprising an Azurocidin signal peptide fused to ectonucleotide pyrophosphatase / phosphodiesterase- 1 (ENPP1) or to ectonucleotide pyrophosphatase / phosphodiesterase-3 (ENPP3), the precursor polypeptide being proteolytically cleaved to produce soluble ENPP1 or soluble ENPP3 which is active to reduce ectopic calcification of soft tissue, upon expression of the polynucleotide in mammalian cells.SELECTED DRAWING: Figure 4