502 results about "Mammalian cell" patented technology

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 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 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.

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.

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.

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.

The invention belongs to the technical field of gene engineering, and particularly relates to application of an Arnt2 gene in improving expression of exogenous recombinant protein of mammalian cells. The nucleotide sequence of the Arnt2 gene is as shown in SEQ ID No.1. The preparation method comprises the following steps: firstly, constructing a recombinant vector for overexpressing the Arnt2, and then transfecting the recombinant vector for overexpressing the Arnt2 into a host cell for expressing recombinant protein to obtain a recombinant cell for stably overexpressing the Arnt2. Experiments show that when the recombinant cell is used for recombinant protein expression, the yield of the recombinant protein is remarkably increased, the expression stability of the recombinant protein is enhanced, and it is indicated that the Arnt2 gene can be used for improving the expression of exogenous recombinant protein of mammalian cells.

This invention provides a recombinant horseshoe crab three-factor composition and its application. The composition comprises recombinant horseshoe crab factor C, recombinant horseshoe crab factor B, and recombinant horseshoe crab coagulase factor; all three factors are expressed by mammalian HEK293 cells. This allows for the expression of factors C, B, and PCE in mammalian HEK293 cells. Simultaneous expression of all three factors enhances factor C activity and improves sensitivity; furthermore, the expressed factors are secretible and possess high activity.

Herein is reported a method for producing a recombinant adeno-associated viral particle preparation (rAAVp), the method comprising the steps of culturing a mammalian cell comprising an expression cassette directed against: a non-adeno-associated viral gene interposed between two AAV inverted terminal repeats (ITRs), and thereby producing the rAAVp; an adeno-associated virus rep gene; an adeno-associated virus cap gene; an adeno-associated virus E1A gene; an adeno-associated virus E1B gene; an adeno-associated virus E2A gene; an adeno-associated virus E4orf6; and an adeno-associated virus VA RNA gene, wherein the culturing is carried out at a pH value in the range of pH 7.4 to pH 7.6 and including a terminal value. The yield of the rAAVp produced by the culture performed at a pH value in the range of pH 7.4 to pH 7.6 and including a terminal value is higher than the yield of the rAAVp produced by the culture performed at a pH value in the range of pH 7.0 to pH 7.2 and including a terminal value, and the yield of the rAAVp produced by the culture performed at a pH value in the range of pH 7.4 to pH 7.6 and including a terminal value is lower than the yield of the rAAVp produced by the culture performed at a pH value in the range of pH 7.0 to pH 7.2. And the rAAVp produced by the culture at a pH value in the range of pH 7.4 to pH 7.6 and including an endvalue has a higher percentage of intact particles than the rAAVp produced by the culture at a pH value in the range of pH 7.0 to pH 7.2 and including an endvalue.

In alternative embodiments, provided are self-activating Förster resonance energy transfer (saFRET) biosensors, and methods for making and using them. In alternative embodiments, provided are self-activating FRET (saFRET) biosensors, and methods that couple FRET and sequencing (FRET-Seq) to integrate random mutagenesis, fluorescence-activated cell sorting (FACS), and next-generation sequencing (NGS) to screen and identify sensitive biosensors from large-scale libraries directly in mammalian cells, utilizing the design of saFRET biosensors as provided herein.

The invention discloses a chikungunya virus envelope E2 protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. The chikungunya virus envelope protein E2 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 chikungunya virus envelope protein E2 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the chikungunya virus-resistant monoclonal antibody with high neutralizing activity and capable of 100% protection of mice against chikungunya virus lethal attack is obtained by carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse toxicity attack experiment after mammalian cell expression and purification, and the recombinant chikungunya virus-resistant monoclonal antibody has characteristics of high neutralizing activity and high immunogenicity, and can be used for preparing the chikungunya virus-resistant monoclonal antibody, and the recombinant chikungunya virus-resistant monoclonal antibody. The monoclonal antibody has application value in prevention and treatment of yellow fever.

A composition is provided comprising a cell population and pharmaceuticals derived from human gingival fibroblasts, present in proportions not found in natural gingival tissue, but having a cellular phenotype preferentially selected to express proteins favorable for angiogenesis and anti-inflammatory effects, while reducing the formation of cell populations that promote tumorigenesis and / or metalloproteinases that inhibit tissue regeneration. A method for producing such a composition that increases proliferation several times over compared to previously known methods is provided, and a method for using such a composition in a wide range of human cell therapies is also provided.

The invention relates to the technical field of gene engineering, in particular to an overexpression vector for improving expression of exogenous recombinant protein of mammalian cells, a recombinant cell and application. The invention also relates to an application of the gene as shown in SEQ ID NO.1 and / or SEQ ID NO.2 in improving expression of exogenous recombinant protein of mammalian cells. The mammalian cell is a CHO cell or an HEK239 cell containing an exogenous recombinant protein. The gene shown in SEQ ID NO.1 is a Ccne1 gene of which the Gene ID is 100753358, and the gene shown in SEQ ID NO.2 is a Ccne1 gene of which the Gene ID The gene shown in the SEQ ID NO.2 is an Aire gene of which the Gene ID is 100768045. The gene shown in the SEQ ID NO.2 is an Aire gene. An overexpression vector of the gene is constructed and transferred into a cell strain containing an exogenous recombinant protein, so that the expression of the exogenous recombinant protein of mammalian cells can be remarkably improved.

The invention relates to a novel human immunodeficiency virus envelope protein antigen expressed by mammals. Various embodiments of the invention relate to a polypeptide comprising 1-10 epitopes or more of the HIV envelope protein and a fusion protein, wherein the polypeptide lacks a transmembrane domain of the HIV gp41 protein. Such polypeptides can be expressed in mammalian cells, such as human cells, to produce polypeptides useful, for example, in the development of novel anti-HIV antibodies. The polypeptides described herein and the novel antibodies developed therefrom are generally useful in medical diagnostics, and they can also be used in the prophylactic and therapeutic treatment of HIV.

To provide an improved antibody-drug conjugate and a pharmaceutical composition containing the antibody-drug conjugate.SOLUTION: The invention is based on the concept that it is possible to generate antibodies that exhibit improved potency (e.g., one or more of increased (e.g., detectable increase) toxin release in target mammalian cells, increased (e.g., detectable increase) killing of target mammalian cells, and increased (e.g., detectable increase) endolysosomal delivery). In some embodiments of any of the antibodies described herein, the heavy chain CH1 - CH2 - CH3 sequence consisting of the specific sequence comprises a substitution of lysine to cysteine at amino acid position 105 and a deletion of threonine at amino acid positions 106 and 108.SELECTED DRAWING: None

Provided herein are methods of selecting an optimal fixed orientation of a dissolved oxygen (DO) probe in a perfusion bioreactor and culturing a mammalian cell in a perfusion bioreactor having the DO probe positioned at the selected fixed orientation.

The invention relates to methods and compositions for preventing or treating a neuropathology in a subject associated with or induced or caused by a P. gingivalis infection, preventing the deposition of or reducing the level of P. gingivalis gingipain in neuronal tissue, delaying the onset of a P. gingivalis-induced or associated neuropathology, for preventing or slowing the rate of abnormal protein deposition in the neuronal tissue, and / or reducing neuroinflammation, the methods comprising administering an RNA polynucleotide encoding a protein comprising or consisting of: - one or more amino acid sequences of an active site of an Arg- or Lys-gingipain of P. gingivalis, or a sequence that is at least 80% identical thereto; and / or - the amino acid sequence of one or more adhesin binding motifs (ABMs) of an adhesin domain of an Arg- or Lys-gingipain of P. gingivalis, or a sequence that is at least 80% identical thereto, wherein the polynucleotide is capable of being translated in a mammalian cell.

An oligonucleotide molecule has 10 to 50 nucleotides that include at least one G-quartet forming motif having 10 to 20 nucleotide residues. At least 60% of the residues of the G-quartet forming motif are guanosine or deoxyguanosine residues. The oligonucleotide molecule inhibits tumor growth and / or viral or bacterial replication and / or exerts anti-inflammatory effects in mammalian cells.

The invention provides a preparation method of a recombinant human blood coagulation factor VIII and a product of the recombinant human blood coagulation factor VIII. The preparation method comprises the step of culturing a mammalian cell strain for expressing the recombinant human blood coagulation factor VIII in a serum-free culture medium of a compound containing copper ions and methionine sulfoximine. By adopting the preparation method provided by the invention, the purity of the recombinant human blood coagulation factor VIII product can be further improved on the basis of keeping the high activity and high yield of the recombinant human blood coagulation factor VIII product obtained by culture.

To provide an IL-15-based molecule that has high stability, an extended in vivo half-life, increased in vivo biological activity, and increased productivity in mammalian cells.SOLUTION: Provided are an immunocytokine comprising a heterodimeric protein complex based on IL-15 / IL-15Rα, and use thereof as a therapeutic agent, in particular as an agent for the treatment of cancer and an autoimmune disease. Provided further are an immunocytokine comprising a heterodimeric protein complex based on an IL-15 / IL-15Rα and an immunomodulatory antibody, and use thereof as a therapeutic agent, in particular as an agent for the treatment of cancer and an autoimmune disease.SELECTED DRAWING: None

The present disclosure relates to methods, cells, and compositions for producing a product of interest, e.g., a recombinant protein. In particular, the present disclosure provides improved mammalian cells expressing the product of interest, where the cells (e.g., Chinese Hamster Ovary (CHO) cells) have reduced or eliminated activity, e.g., expression, of certain host cell proteins, e.g., enzymes including, but not limited to, certain lipases, esterases, and / or hydrolases.

Provided herein are antigen-binding protein constructs capable of specifically binding FOLR1 or an epitope of FOLR1 presented on the surface of a target mammalian cell, wherein said antigen binding is pH-dependent. Provided are also uses of said antigen-binding protein constructs.

The invention relates to the technical field of biology, in particular to a screening system and a screening method for transcription factor binding targets. The invention provides a system and a method for screening transcription factor binding targets based on rolling circle amplification (RCA) by optimizing a template structure in an existing rolling circle amplification system, increasing auxiliary reactants and regulating RCA amplification time, template initial concentration and PCR template sample loading amount. The screening system and the screening method can respond to screening requirements of transcription factor weak binding targets, a detection signal is amplified through a synergistic binding effect of multiple sequences, the basic background noise of a constructed library is low, good compatibility is shown in a mammalian cell screening system, obvious cytotoxicity is not caused, and the screening system and the screening method have good application prospects. The library sequence diversity is controllable, and the stability is high; compared with traditional single sequence screening, the method has the advantages that the detection rate of weak binding targets is remarkably increased, and false negative results can be effectively reduced.

The invention discloses a yellow fever virus envelope protein monoclonal antibody and application thereof, and belongs to the technical field of medicines. Yellow fever virus envelope protein E 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 yellow fever virus envelope protein E are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. According to the present invention, the monoclonal antibody with high neutralizing activity and capable of completely protecting mice from yellow fever virus lethal attack is obtained by using the monoclonal antibody as a template, connecting the monoclonal antibody and the constant region gene to an expression vector, carrying out mammalian cell expression and purification, and carrying out enzyme-linked immunosorbent assay, in-vitro virus neutralization and mouse virus attack experiment, such that the monoclonal antibody has high neutralizing activity and can completely protect mice from yellow fever virus lethal attack; the monoclonal antibody has application value in prevention and treatment of yellow fever.