公开(公告)号：US06919183B2

公开(公告)日：2005-07-19

申请号：US10050279

申请日：2002-01-16

申请人： James P. Fandl ,  Neil Stahl ,  Gang Chen ,  George D. Yancopoulos

发明人： James P. Fandl ,  Neil Stahl ,  Gang Chen ,  George D. Yancopoulos

IPC分类号： A01K67/027 ,  C07K14/315 ,  C07K14/52 ,  C07K16/28 ,  C12N5/10 ,  C12N15/09 ,  C40B30/04 ,  G01N33/569 ,  G01N33/68 ,  G01N33/53 ,  C12N21/02 ,  C12N21/08

CPC分类号： C40B30/04 ,  A01K2217/05 ,  A01K2267/01 ,  C07K14/315 ,  C07K14/52 ,  C07K16/2863 ,  C07K2317/77 ,  C07K2319/00 ,  C12N15/1051 ,  G01N33/56966 ,  G01N33/6845 ,  G01N2500/10

摘要： A method for identifying and isolating cells which produce secreted proteins. This method is based upon a specific characteristic or the expression level of the secreted protein by transiently capturing the secreted protein on the surface of an individual cell, allowing selection of rare cell clones from a heterogeneous population. Also provided is the use of this method to generate cells which produce a desired level of secreted protein or secreted protein of a particular characteristic(s), and organisms which possess such cells. In particular, the method allows rapid isolation of high expression recombinant antibody-producing cell lines, or may be applied directly to rapid isolation of specific hybridomas, or to the isolation of antibody-producing transgenic animals. This method is applicable for any cell which secretes protein.

摘要翻译： 用于鉴定和分离产生分泌蛋白的细胞的方法。 该方法基于通过瞬时捕获单个细胞表面上的分泌蛋白质的分泌蛋白质的特定特征或表达水平，允许从异质群体中选择稀有细胞克隆。 还提供了这种方法用于产生产生具有特定特征的所需水平的分泌蛋白质或分泌蛋白质的细胞以及具有这种细胞的生物体的用途。 特别地，该方法允许高表达重组抗体产生细胞系的快速分离，或者可以直接应用于快速分离特异性杂交瘤，或分离产生抗体的转基因动物。 该方法适用于分泌蛋白质的任何细胞。

公开(公告)号：US08673589B2

公开(公告)日：2014-03-18

申请号：US12323161

申请日：2008-11-25

申请人： Gang Chen ,  Changlin Dou ,  James P. Fandl

发明人： Gang Chen ,  Changlin Dou ,  James P. Fandl

IPC分类号： C12P21/06 ,  C12N15/74

CPC分类号： C12N15/635 ,  A01K2217/05 ,  C07K16/00 ,  C07K2317/14 ,  C07K2317/21 ,  C07K2319/00 ,  C12N15/63 ,  C12N15/85 ,  C12N2830/00 ,  C12N2830/15 ,  C12N2830/42 ,  C12N2840/20 ,  C12P21/00

摘要： Compositions and methods for the inducible expression of genes in eukaryotic cells are provided. Expression of a nucleotide sequence of interest encoding a protein of interest is controlled by a regulatory fusion protein that consists of a transcription blocking domain and a ligand-binding domain. When a cognate ligand for the ligand-binding domain is present, transcription of the nucleotide sequence of interest is blocked. Upon removal of the cognate ligand, the nucleotide sequence of interest is transcribed. The method is useful for large scale bioreactor production of a desired protein of interest in eukaryotic cells.

摘要翻译： 提供了在真核细胞中可诱导表达基因的组合物和方法。 通过由转录阻断结构域和配体结合结构域组成的调节融合蛋白来控制感兴趣的蛋白质的目标核苷酸序列的表达。 当存在配体结合结构域的同源配体时，感兴趣的核苷酸序列的转录被阻断。 去除同源配体后，转录目的核苷酸序列。 该方法可用于在真核细胞中大规模生物反应器生产期望的蛋白质。

公开(公告)号：US20100304436A1

公开(公告)日：2010-12-02

申请号：US12791637

申请日：2010-06-01

申请人： Gang CHEN ,  Darya Burakov ,  James P. Fandl

发明人： Gang CHEN ,  Darya Burakov ,  James P. Fandl

IPC分类号： C12P21/00 ,  C07H21/04 ,  C12N5/10

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。

公开(公告)号：US07771997B2

公开(公告)日：2010-08-10

申请号：US12132846

申请日：2008-06-04

申请人： Gang Chen ,  Robert Babb ,  James P. Fandl

发明人： Gang Chen ,  Robert Babb ,  James P. Fandl

IPC分类号： C12N5/10

CPC分类号： C12N15/85 ,  C07K16/00 ,  C07K2319/30 ,  C12N15/86 ,  C12N2740/10043 ,  C12N2800/30 ,  C12N2840/203 ,  C12P21/02

摘要： Expression-enhancing nucleotide sequences for expression in eukaryotic systems are provided that allow for enhanced and stable expression of recombinant proteins in eukaryotic cells. Enhanced expression and stability regions (EESYRs) 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.

摘要翻译： 提供了用于在真核系统中表达的表达增强核苷酸序列，其允许重组蛋白在真核细胞中增强和稳定表达。 提供增强的表达和稳定区（EESYR）用于在真核细胞中表达感兴趣的基因。 提供染色体基因座，序列和载体用于真核细胞中基因的增强和稳定表达。

公开(公告)号：US20090137416A1

公开(公告)日：2009-05-28

申请号：US12240541

申请日：2008-09-29

申请人： James P. Fandl ,  Gang Chen ,  Neil Stahl ,  George D. Yancopoulos

发明人： James P. Fandl ,  Gang Chen ,  Neil Stahl ,  George D. Yancopoulos

IPC分类号： C40B30/04 ,  G01N33/53

CPC分类号： G01N33/6854 ,  C12N15/65 ,  G01N33/566 ,  G01N33/56966 ,  G01N2333/7051

摘要： A method of detecting and isolating cells that produce a secreted protein of interest (POI) that has a T cell receptor variable 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.

摘要翻译： 一种检测和分离产生具有T细胞受体可变区的目标分泌蛋白质（POI）的细胞的方法，包括：a）通过以下步骤构建细胞系：瞬时或稳定表达结合POI的细胞表面捕获分子，通过 用编码这种细胞表面捕获分子的核酸转染细胞系; b）同时或随后用编码POI的第二核酸同时转染所述细胞，其中所述POI被分泌; c）通过使细胞与结合POI的检测分子接触来检测表面显示的POI; 和d）基于检测分子分离细胞。

公开(公告)号：US06472178B1

公开(公告)日：2002-10-29

申请号：US09031693

申请日：1998-02-27

申请人： James P. Fandl ,  Neil E. Stahl ,  Stanley J. Wiegand

发明人： James P. Fandl ,  Neil E. Stahl ,  Stanley J. Wiegand

IPC分类号： C12N1518

CPC分类号： G01N33/6842 ,  A61K38/185 ,  C07K14/475 ,  G01N33/68

摘要： Modified ciliary neurotrophic factor, methods for production and methods of use, especially in the treatment of Huntington's disease, obesity, and gestational or adult onset diabetes.

摘要翻译： 改良的睫状神经营养因子，生产方法和使用方法，特别是治疗亨廷顿病，肥胖症和妊娠或成人发病的糖尿病。

公开(公告)号：US08389239B2

公开(公告)日：2013-03-05

申请号：US12793898

申请日：2010-06-04

申请人： Gang Chen ,  Robert Babb ,  James P. Fandl

发明人： Gang Chen ,  Robert Babb ,  James P. Fandl

IPC分类号： C12P21/02

CPC分类号： C12N15/85 ,  C07K16/00 ,  C07K2319/30 ,  C12N15/86 ,  C12N2740/10043 ,  C12N2800/30 ,  C12N2840/203 ,  C12P21/02

摘要： Expression-enhancing nucleotide sequences for expression in eukaryotic systems are provided that allow for enhanced and stable expression of recombinant proteins in eukaryotic cells. Enhanced expression and stability regions (EESYRs) 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.

公开(公告)号：US20100291626A1

公开(公告)日：2010-11-18

申请号：US12793898

申请日：2010-06-04

申请人： Gang CHEN ,  Robert Babb ,  James P. Fandl

发明人： Gang CHEN ,  Robert Babb ,  James P. Fandl

IPC分类号： C12P21/00 ,  C12N5/10

CPC分类号： C12N15/85 ,  C07K16/00 ,  C07K2319/30 ,  C12N15/86 ,  C12N2740/10043 ,  C12N2800/30 ,  C12N2840/203 ,  C12P21/02

摘要： Expression-enhancing nucleotide sequences for expression in eukaryotic systems are provided that allow for enhanced and stable expression of recombinant proteins in eukaryotic cells. Enhanced expression and stability regions (EESYRs) 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.

摘要翻译： 提供了用于在真核系统中表达的表达增强核苷酸序列，其允许重组蛋白在真核细胞中增强和稳定表达。 提供增强的表达和稳定区（EESYR）用于在真核细胞中表达感兴趣的基因。 提供染色体基因座，序列和载体用于真核细胞中基因的增强和稳定表达。

公开(公告)号：US20090162901A1

公开(公告)日：2009-06-25

申请号：US12323161

申请日：2008-11-25

申请人： Gang Chen ,  Changlin Dou ,  James P. Fandl

发明人： Gang Chen ,  Changlin Dou ,  James P. Fandl

IPC分类号： C12P21/00

CPC分类号： C12N15/635 ,  A01K2217/05 ,  C07K16/00 ,  C07K2317/14 ,  C07K2317/21 ,  C07K2319/00 ,  C12N15/63 ,  C12N15/85 ,  C12N2830/00 ,  C12N2830/15 ,  C12N2830/42 ,  C12N2840/20 ,  C12P21/00

摘要： Compositions and methods for the inducible expression of genes in eukaryotic cells are provided. Expression of a nucleotide sequence of interest encoding a protein of interest is controlled by a regulatory fusion protein that consists of a transcription blocking domain and a ligand-binding domain. When a cognate ligand for the ligand-binding domain is present, transcription of the nucleotide sequence of interest is blocked. Upon removal of the cognate ligand, the nucleotide sequence of interest is transcribed. The method is useful for large scale bioreactor production of a desired protein of interest in eukaryotic cells.

摘要翻译： 提供了在真核细胞中可诱导表达基因的组合物和方法。 通过由转录阻断结构域和配体结合结构域组成的调节融合蛋白来控制感兴趣的蛋白质的目标核苷酸序列的表达。 当存在配体结合结构域的同源配体时，感兴趣的核苷酸序列的转录被阻断。 去除同源配体后，转录目的核苷酸序列。 该方法可用于在真核细胞中大规模生物反应器生产期望的蛋白质。

公开(公告)号：US07534604B2

公开(公告)日：2009-05-19

申请号：US11035599

申请日：2005-01-14

申请人： James P. Fandl ,  Gang Chen ,  Nicholas Papadopoulos ,  Thomas H. Aldrich

发明人： James P. Fandl ,  Gang Chen ,  Nicholas Papadopoulos ,  Thomas H. Aldrich

IPC分类号： C12P21/04 ,  C12N5/00 ,  C07H21/04

CPC分类号： C07K16/2863 ,  C07H21/04 ,  C07K2317/31 ,  C07K2317/52 ,  C07K2317/622 ,  C07K2317/75 ,  C07K2319/30

摘要： A fusion polypeptide comprising (A)x-M-(A′)y, wherein A and A′ are each polypeptides capable of binding a target receptor. The fusion polypeptides of the invention form multimeric proteins which activate the target receptor. A and A′ may be each be an antibody or fragment derived from an antibody specific for a target receptor, such as the same or different ScFv fragments, and/or a ligand or ligand fragment or derivative capable of binding the target protein, M is a multimerizing component, and X and Y are independently a number between 1-10.

摘要翻译： 包含（A）x-M-（A'）y的融合多肽，其中A和A'是能够结合靶受体的多肽。 本发明的融合多肽形成激活靶受体的多聚体蛋白。 A和A'可以各自为衍生自靶受体（例如相同或不同ScFv片段）的特异性抗体的抗体或片段，和/或能够结合靶蛋白质的配体或配体片段或衍生物，M为 多聚化组分，X和Y独立地为1-10之间的数。