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公开(公告)号：US20240018553A1
公开(公告)日：2024-01-18
申请号：US18461798
申请日：2023-09-06
发明人： Ying Shen , Darya Burakov , Gang Chen , James P. Fandl
CPC分类号： C12N15/907 , C12N15/85
摘要： Expression-enhancing nucleotide sequences for eukaryotic expressions systems are provided that allow for enhanced and stable expression of recombinant proteins in eukaryotic cells. Genomic integration sites providing enhanced expression and methods of use thereof are provided for expression of a gene of interest in a eukaryotic cell. Chromosomal loci, sequences, and vectors are provided for enhanced and stable expression of genes in eukaryotic cells.
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公开(公告)号：US10351622B2
公开(公告)日：2019-07-16
申请号：US15624982
申请日：2017-06-16
发明人： Gang Chen , Darya Burakov , Dipali Deshpande
IPC分类号： C07K16/22 , C12N15/85 , C07K14/47 , C12N15/79 , C07K16/00 , C12N9/24 , C12N15/09 , C12N15/63
摘要： The present invention relates to discovery of the ectopic expression of EDEM2 in a production cell to improve the yield of a useful multi-subunit protein. Thus, the present invention provides for production cell lines, such as the canonical mammalian biopharmaceutical production cell, the CHO cell, containing recombinant polynucleotides encoding EDEM2. Also disclosed is a production cell containing both an EDEM2-encoding polynucleotide as well an XBP1-encoding polynucleotide. Improved titers of antibodies produced by these cell lines are disclosed, as well as the improved cell densities attained by these cells in culture.
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公开(公告)号：US09206455B2
公开(公告)日：2015-12-08
申请号：US13779952
申请日：2013-02-28
发明人： Gang Chen , Darya Burakov , James P. Fandl
CPC分类号： C12P21/005 , C07K16/00 , C07K16/2863 , C07K16/2866 , C07K2317/14 , C07K2317/41 , C12N9/0006 , C12N15/00 , C12Y101/01271 , Y02P20/52
摘要： An isolated nucleic acid encoding an FX protein having a serine at position 79, a lysine at position 90, a leucine at position 136, an arginine at position 211, a serine at position 289, and a combination thereof is provided. Cells having a gene encoding a modified FX protein are provided, wherein the cells exhibit a reduced ability to fucosylate a glycoprotein at a first temperature, but exhibit the ability to fucosylate the glycoprotein at a second temperature. Methods and compositions for making glycoproteins with reduced fucosylation are provided.
摘要翻译： 提供了编码FX蛋白的分离核酸，其在79位具有丝氨酸，在90位具有赖氨酸，136位的亮氨酸，211位的精氨酸，289位的丝氨酸及其组合。 提供具有编码经修饰的FX蛋白的基因的细胞，其中细胞在第一温度下表现出降低糖蛋白岩藻糖化能力，但在第二温度下表现出对糖蛋白的岩藻糖化能力。 提供了用于制备具有降低的岩藻糖基化的糖蛋白的方法和组合物。
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公开(公告)号：US12054751B2
公开(公告)日：2024-08-06
申请号：US18066303
申请日：2022-12-15
发明人： Gang Chen , Darya Burakov , James P. Fandl
CPC分类号： C12N9/0006 , C12N15/00 , C12P21/005 , C12Y101/01271 , C07K16/00 , C07K16/2863 , C07K16/2866 , C07K2317/14 , C07K2317/41 , Y02P20/52
摘要： An isolated nucleic acid encoding an FX protein having a serine at position 79, a lysine at position 90, a leucine at position 136, an arginine at position 211, a serine at position 289, and a combination thereof is provided. Cells having a gene encoding a modified FX protein are provided, wherein the cells exhibit a reduced ability to fucosylate a glycoprotein at a first temperature, but exhibit the ability to fucosylate the glycoprotein at a second temperature. Methods and compositions for making glycoproteins with reduced fucosylation are provided.
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公开(公告)号：US20230304062A1
公开(公告)日：2023-09-28
申请号：US18047349
申请日：2022-10-18
发明人： Yu Zhao , Michael Goren , Darya Burakov , Gang Chen
CPC分类号： C12P21/02 , C12N5/00 , C12N15/86 , C12N2750/14143 , C12N2830/002
摘要： Compositions and methods for the transcription of polynucleotide sequences encoding polypeptides in cells are provided. The cells and methods provided are useful for controlling expression of polypeptides for a wide variety of purposes.
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公开(公告)号：US20210340564A1
公开(公告)日：2021-11-04
申请号：US17366239
申请日：2021-07-02
发明人： Dipali Deshpande , Darya Burakov , Gang Chen , James Fandl
摘要： The invention provides a new expression system comprising a mammalian selectable marker that promotes desirable post-translational modifications of glycoproteins. In particular, the invention includes methods and compositions for optimal recombinant protein expression in mammalian cells by employing a selection marker system based on GPT genes of mammalian origin. The invention includes methods that facilitate selectivity and enhanced expression copies as well as protein yield of recombinant proteins in mammalian cells, and methods of using GPT expression systems.
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公开(公告)号：US20190270800A1
公开(公告)日：2019-09-05
申请号：US16417792
申请日：2019-05-21
发明人： Gang Chen , Darya Burakov , Dipali Deshpande
摘要： The present invention relates to discovery of the ectopic expression of EDEM2 in a production cell to improve the yield of a useful multi-subunit protein. Thus, the present invention provides for production cell lines, such as the canonical mammalian biopharmaceutical production cell, the CHO cell, containing recombinant polynucleotides encoding EDEM2. Also disclosed is a production cell containing both an EDEM2-encoding polynucleotide as well an XBP1-encoding polynucleotide. Improved titers of antibodies produced by these cell lines are disclosed, as well as the improved cell densities attained by these cells in culture.
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公开(公告)号：US20170241887A1
公开(公告)日：2017-08-24
申请号：US15443290
申请日：2017-02-27
发明人： James P. Fandl , Gang Chen , Neil Stahl , George D. Yancopoulos , Dipali Deshpande , Darya Burakov , Thomas Aldrich , Vishal Kamat
IPC分类号： G01N15/14 , G01N33/569 , C07K16/00 , G01N33/68
CPC分类号： G01N15/14 , C07K16/00 , C07K16/2863 , C07K16/4283 , C07K2317/31 , C07K2317/33 , C07K2317/52 , C07K2317/526 , C07K2317/54 , C07K2317/565 , C07K2317/622 , C07K2317/92 , C07K2317/94 , C07K2319/74 , G01N33/56966 , G01N33/68 , G01N2015/0065 , G01N2015/1006 , G01N2015/149
摘要： A method of detecting and isolating cells that produce a secreted protein of interest (POI) that has an immunoglobulin CH3 domain and/or substituted CH3 domain, comprising: a) constructing a cell line transiently or stably expressing a cell surface capture molecule, which binds the POI, by transfecting the cell line with a nucleic acid that encodes such cell surface capture molecule; b) transfecting said cell simultaneously or subsequently with a second nucleic acid that encodes a POI wherein such POI is secreted; c) detecting the surface-displayed POI by contacting the cells with a detection molecule, which binds the POI; and d) isolating cells based on the detection molecule.
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公开(公告)号：US09193786B2
公开(公告)日：2015-11-24
申请号：US14754112
申请日：2015-06-29
发明人： Gang Chen , Darya Burakov , Dipali Deshpande
CPC分类号： C07K16/22 , C07K14/47 , C07K14/4705 , C07K16/00 , C07K2317/14 , C07K2317/51 , C07K2317/515 , C12N9/2488 , C12N15/09 , C12N15/63 , C12N15/79 , C12N15/85 , C12N2015/8518 , C12Y302/01
摘要： The present invention relates to discovery of the ectopic expression of EDEM2 in a production cell to improve the yield of a useful multi-subunit protein. Thus, the present invention provides for production cell lines, such as the canonical mammalian biopharmaceutical production cell—the CHO cell, containing recombinant polynucleotides encoding EDEM2. Also disclosed is a production cell containing both an EDEM2-encoding polynucleotide as well an XBP1-encoding polynucleotide. Improved titers of antibodies produced by these cell lines are disclosed, as well as the improved cell densities attained by these cells in culture.
摘要翻译： 本发明涉及在生产细胞中发现EDEM2的异位表达以改善有用的多亚单位蛋白的产量。 因此，本发明提供了生产细胞系，例如含有编码EDEM2的重组多核苷酸的规范哺乳动物生物制药生产细胞 - CHO细胞。 还公开了含有EDEM2编码多核苷酸以及编码XBP1的多核苷酸两者的生产细胞。 公开了由这些细胞系产生的抗体的改善的滴度，以及培养中由这些细胞获得的改善的细胞密度。
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公开(公告)号：US11788102B2
公开(公告)日：2023-10-17
申请号：US16575506
申请日：2019-09-19
发明人： Ying Shen , Darya Burakov , Gang Chen , James P. Fandl
CPC分类号： C12N15/907 , C12N15/85
摘要： Expression-enhancing nucleotide sequences for eukaryotic expressions systems are provided that allow for enhanced and stable expression of recombinant proteins in eukaryotic cells. Genomic integration sites providing enhanced expression and methods of use thereof are provided for expression of a gene of interest in a eukaryotic cell. Chromosomal loci, sequences, and vectors are provided for enhanced and stable expression of genes in eukaryotic cells.
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