284 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 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.

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.

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 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 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.

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.

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.

This specification provides an improved method for producing Fc-containing proteins. The method generally comprises culturing mammalian cells that produce Fc-containing proteins at a first pH setting for a first period of time, and then culturing the cells at a second pH setting higher than the first pH setting for a second period of time.

Provided herein are engineered or modified cell lines in which the expression of at least one of ADM2, AURKA, BCL2, CCND1, CHD1, CREB3L3, RIGI / DDX58, ECSIT, EHHADH, ERBB2, FAS, FBXW7, FGFR3, FOXL2, GATA1, HDAC5, HOXA5, HSP90B1, IL6, ITGA2B, KAT5, LAS1L, LIF, NFKB1, NFKB2, NFKB1A, NFE2L2, NGEF, NME1, NR3C1, POLN, PPAR-A, PTEN, RELB, RPA1, RXRA, SMAD4, SS18, TCF7, TGM2, THEM6, TNFAIP3, TP63, TRPC1, DBH, ITGAM, HHEX, TXNIP, ASNS, PLEKH02, CDKN1A, C5orf64, COL20A1, GJB3, ISYNA1, PARD6G-AS1, ACTA1, ACTC1, ADCY10, ADCY4, ADORA2A, ALKBH5, ATP1A4, ATP1B2, ATP2B2, BAIAP3, BAZ1B, BTK, CBL, CCNA1, CD14, CLTC, COPS3, CRHR2, CXADR, DAPK1, DCTN5, DYNLRB1, EME2, EXOC3L1, EYA3, EYA4, FANCA, FEN1, GAK, GCGR, GHSR, GSK3B, H2BC5, H4C7, H4C8, HAP1, HCAR1, INO80, KCNB1, LAT2, LIN7C, MAP1B, MBD4, MYH11, MYH15, MYH3, MYH7B, NAPA, NKD2, NLRP5, NPM2, NR4A3, NFRKB, PAXIP1, PIAS1, PIAS4, POLE, POLE3, POLD1, POLH, PPP4R2, PTGER3, RAB25, RAB40C, RAB44, RAD9A, REV1, RIF1, RNF111, RNF168, RNF4, RPA4, RTEL1, SCRIB, SNRNP70, SPHK1, STX3, SYT1, SYT2, SYT5, SYTL1, SYNJ2, TERF2, TNFSF10, TNPO2, TOP3A, TRIM25, TRIM72, TXLNA, UBA7, UNC5A, VAV3, VPS18, WASL, XRN2, and YY1 is altered as compared to a control cell line. Also provided are methods of producing recombinant AAV particles in said engineered or modified cell lines.

An engineered tyrosyl-tRNA synthetase / tRNA pair that co-translationally incorporates O-sulfotyrosine in response to UAG codons in E. coli and mammalian cells is described herein. This platform enables recombinant expression of eukaryotic proteins homogeneously sulfated at chosen sites.

The inventions provide mammalian cell lines integrated with recombinant adeno-associated virus (rAAV) plasmids, cell genomes and methods of making the cell genomes for production of rAAV. The inventions also provide regulatory fusion proteins (RFPs) (which can act as activators or repressors) and repressor proteins (which act as repressors), such as antibiotic repressors, and arranged operators to control transcription of at least one polynucleotide of interest, such as AAV Rep genes and / or non-AAV helper genes.

The present invention describes methods for obtaining mammalian cell lines that express recombinant equine chorionic gonadotropin (reCG) hormone. Cell lines expressing reCG, large-scale methods for producing reCG, reCG with higher biological activity relative to PMSG, formulations containing reCG, nucleic acids encoding reCG, and uses are also described.

Herein is reported a method for separating recombinant viral particles from other compounds of a mammalian cell culture broth comprising the steps of adding a silica-based filter aid like diatomaceous earth (DE) or synthetic silica-based filter aid to a mammalian cell culture broth comprising recombinant viral particles at a weight ratio of more than 1:2 DE or synthetic silica-based filter aid / biological wet mass (BWM) to obtain a pre-filtration mixture and subjecting the pre-filtration mixture to alluvial filtration, whereby recombinant viral particles are separated from other compounds of the mammalian cell culture broth.

The present invention relates to a method for identifying a compound that modulates the mitotic cell cycle in cancer cells by modulating at least one of the expression, the amount, the stability and / or the biological activity of the protein CITED2 and / or CITED2 containing protein complexes in a mammalian cell. Further provided is a method for determining the level of CITED2 and / or CDK11bp58 and / or splicing factor 3B subunit 1 (SF3B1), and / or 14-3-3γ in a sample obtained from a subject, comprising detecting of binding of CITED2 in the sample to at least one molecule selected from the group consisting of CDK11bp58, splicing factor 3B subunit 1 (SF3B1), and 14-3-3γ, wherein a decrease of the binding, when compared to a control indicates an improper centrosome segregation and subsequent an increase of the formation of chaotic monopolar mitotic spindles in said sample. Further provided are pharmaceutical compositions comprising the compound as identified for use in medicine, in particular for use in the prevention or treatment of a disease or condition caused by cancer.

Dual vector systems and these comprehensive mammalian host cells are provided for the expression of multi-chain molecules, with each vector employing a different promoter controlling the expression of a selectable marker. The mammalian host cells can be used to generate a recombinant protein of interest in a mammalian cell culture.

Described herein are artificial centromeres and artificial chromosomes comprising the artificial centromeres and a cargo sequence, methods of designing and making such artificial centromeres and artificial chromosomes, cells comprising such artificial centromeres and artificial chromosomes, and methods of using such artificial centromeres and artificial chromosomes for stable passaging of artificial chromosomes and expression of cargo sequences in a mammalian cell.

The invention provides a skin organoid construction method of human embryonic stem cells based on high-expression recombinant human COL17A1 protein and application of the skin organoid construction method, and relates to the technical field of molecular biology, and the skin organoid construction method comprises the following steps: integrating nucleic acid molecules for coding COL17A1 protein to a KRT5 site of genome DNA of the human embryonic stem cells to obtain recombinant human embryonic stem cells; the nucleic acid molecule for coding the COL17A1 protein contains the gene for coding the COL17A1 protein, and compared with the traditional'random integration 'or'universal safe port site' (AAVS1), the yield of the recombinant human COL17A1 protein is remarkably improved by integrating at the KRT5 site, and the stable and efficient expression of the recombinant human COL17A1 protein is realized. The technical problem that in the prior art, the expression quantity of recombinant human COL17A1 protein is low when mammalian cells such as HEK-293T and CHO are used for expressing the recombinant human COL17A1 protein is solved.

This invention provides materials and methods for regulating ligand-gated ion channel (LGIC) activity. [Solution] A modified LGIC is provided, comprising at least one LGIC subunit having a modified ligand-binding domain (LBD) and / or a modified ion-pore domain (IPD). Also provided are an exogenous LGIC ligand capable of binding to and activating the modified LGIC, as well as a method for regulating ion transport across the membrane of mammalian cells, a method for regulating cellular excitability in mammals, and a method for treating mammals with channel disorders.

Dual vector systems and mammalian host cells comprising these dual vector systems are provided, wherein each vector employs a different promoter that drives the expression of a selectable marker. A mammalian host cell can be used to produce a recombinant protein of interest in mammalian cell culture.

The present invention relates to a serum-free cell culture perfusion medium comprising medium components that make up at least three separate aqueous concentrated feeds and a diluent wherein the resulting serum-free cell culture perfusion medium, upon mixing, adjusts the pH to a neutral pH. The invention also provides a method for preparing the serum-free cell culture perfusion medium. The invention further relates to methods of culturing mammalian cells or producing proteins of interest in perfusion cultures using the serum-free cell culture perfusion media, which achieve high productivity at low cell specific perfusion rates. The invention further relates to the use of novel and improved serum-free cell culture perfusion media to control the osmotic pressure of a perfusion cell culture, where an increase in total productivity and / or cell specific productivity is caused by inhibiting cell growth, increasing osmotic pressure during cell culture, for example during the production phase of perfusion cell culture. Inhibiting cell growth particularly reduces or eliminates the need for wasted cell expulsion.

The invention belongs to the technical field of biology, and discloses a polypeptide derived from a deep sea ecological environment, an application of the polypeptide in preparation of an antibacterial preparation and an antibacterial drug. According to the application of the polypeptide disclosed by the invention in the preparation of the antibacterial preparation, the polypeptide comprises one or more of amino acid fragments with sequences shown as SEQ ID NO: 1-9, the compound has good bacteriostatic or bactericidal activity on one or more of escherichia coli, halomonas aeruginosa, acinetobacter picoteri, staphylococcus aureus, linear preristeria, klebsiella pneumoniae and vibrio parahaemolyticus, and has extremely low toxicity on mammalian cells under an effective concentration. The compound has excellent bacteriostatic activity, bacteriostatic broad-spectrum property and biocompatibility, and has very excellent application prospects in multiple fields such as food preservation, agricultural disease prevention and control, feed additives and the like.