公开(公告)号：US20130084593A1

公开(公告)日：2013-04-04

申请号：US13561395

申请日：2012-07-30

申请人： Yen-Tung Luan ,  Wenge Wang ,  Ryan Nolan ,  Denis Drapeau

发明人： Yen-Tung Luan ,  Wenge Wang ,  Ryan Nolan ,  Denis Drapeau

IPC分类号： C12Q1/02

CPC分类号： C12Q1/02 ,  C12N5/0018 ,  C12N2500/32

摘要： This invention relates to methods for rationally designing cell culture media for use in cell cultures, e.g., cell cultures employed in polypeptide production; cell culture media designed with the disclosed methods; methods of producing a polypeptide of interest, e.g., an antibody, using such media; polypeptides produced using the methods and media disclosed herein; and pharmaceuticals compositions containing such polypeptides. The rationally designed media contain a concentration of an amino acid that is calculated for use in cell mass, a concentration of the amino acid that is calculated for use in cell maintenance, and a concentration of the amino acid that is calculated for incorporation into the polypeptide of interest.

摘要翻译： 本发明涉及用于合理设计用于细胞培养物的细胞培养基的方法，例如用于多肽生产的细胞培养物; 用公开的方法设计的细胞培养基; 使用这种培养基生产感兴趣多肽的方法，例如抗体; 使用本文公开的方法和介质产生的多肽; 和含有这种多肽的药物组合物。 合理设计的培养基含有计算用于细胞质量的氨基酸浓度，计算用于细胞维持的氨基酸的浓度以及计算用于掺入多肽的氨基酸的浓度 出于兴趣。

公开(公告)号：US20080108553A1

公开(公告)日：2008-05-08

申请号：US11936866

申请日：2007-11-08

申请人： Yen-Tung Luan ,  Wenge Wang ,  Ryan Nolan ,  Denis Drapeau

发明人： Yen-Tung Luan ,  Wenge Wang ,  Ryan Nolan ,  Denis Drapeau

IPC分类号： A61K38/02 ,  C12P21/00 ,  C12N5/06 ,  C07K2/00

CPC分类号： C12Q1/02 ,  C12N5/0018 ,  C12N2500/32

摘要： This invention relates to methods for rationally designing cell culture media for use in cell cultures, e.g., cell cultures employed in polypeptide production; cell culture media designed with the disclosed methods; methods of producing a polypeptide of interest, e.g., an antibody, using such media; polypeptides produced using the methods and media disclosed herein; and pharmaceuticals compositions containing such polypeptides. The rationally designed media contain a concentration of an amino acid that is calculated for use in cell mass, a concentration of the amino acid that is calculated for use in cell maintenance, and a concentration of the amino acid that is calculated for incorporation into the polypeptide of interest. The rationally designed media may contain a baseline-adjusted concentration, A, of at least one amino acid that is calculated according to the formula A=[(M*X)+(N*P)+(Y*M*X)]*F, wherein X is the concentration of the amino acid that is used per unit of cell mass, P is the concentration of the amino acid that is used for incorporation into the polypeptide of interest per unit of polypeptide titer, M is the multiplier for the desired peak cell density of the cell culture, N is the multiplier for the desired concentration of the polypeptide of interest, Y is the cell maintenance factor; and F is the baseline factor. This rationally designed media may also be used to produce a starting cell culture medium comprising a concentration, B, of at least one amino acid according to the formula B=[A−(Z*V)]/(1−V), wherein Z is a concentration of the amino acid in the feeding cell culture medium, and V is a volume of the feeding culture medium as a proportion of the desired cell culture medium volume.

摘要翻译： 本发明涉及用于合理设计用于细胞培养物的细胞培养基的方法，例如用于多肽生产的细胞培养物; 用公开的方法设计的细胞培养基; 使用这种培养基生产感兴趣多肽的方法，例如抗体; 使用本文公开的方法和介质产生的多肽; 和含有这种多肽的药物组合物。 合理设计的培养基含有计算用于细胞质量的氨基酸浓度，计算用于细胞维持的氨基酸的浓度以及计算用于掺入多肽的氨基酸的浓度 出于兴趣。 合理设计的培养基可以含有根据公式A = [（M * X）+（N * P）+（Y * M * X）]计算的至少一个氨基酸的基线调整浓度A * F，其中X是每单位细胞质量使用的氨基酸的浓度，P是单位多肽滴度用于掺入感兴趣多肽的氨基酸的浓度，M是 细胞培养物所需的峰值细胞密度，N是感兴趣多肽所需浓度的乘数，Y是细胞维持因子; F是基线因子。 这种合理设计的培养基也可用于产生包含至少一种根据式B = [A-（Z * V）] /（1-V）的氨基酸的浓度B的起始细胞培养基，其中 Z是饲养细胞培养基中氨基酸的浓度，V是作为所需细胞培养基体积的比例的饲养培养基的体积。

公开(公告)号：US08232075B2

公开(公告)日：2012-07-31

申请号：US11936866

申请日：2007-11-08

申请人： Yen-Tung Luan ,  Wenge Wang ,  Ryan Nolan ,  Denis Drapeau

发明人： Yen-Tung Luan ,  Wenge Wang ,  Ryan Nolan ,  Denis Drapeau

IPC分类号： C12P21/00 ,  C12N5/00 ,  A61K38/02

CPC分类号： C12Q1/02 ,  C12N5/0018 ,  C12N2500/32

摘要： This invention relates to methods for rationally designing cell culture media for use in cell cultures, e.g., cell cultures employed in polypeptide production; cell culture media designed with the disclosed methods; methods of producing a polypeptide of interest, e.g., an antibody, using such media; polypeptides produced using the methods and media disclosed herein; and pharmaceuticals compositions containing such polypeptides. The rationally designed media contain a concentration of an amino acid that is calculated for use in cell mass, a concentration of the amino acid that is calculated for use in cell maintenance, and a concentration of the amino acid that is calculated for incorporation into the polypeptide of interest.

摘要翻译： 本发明涉及用于合理设计用于细胞培养物的细胞培养基的方法，例如用于多肽生产的细胞培养物; 用公开的方法设计的细胞培养基; 使用这种培养基生产感兴趣多肽的方法，例如抗体; 使用本文公开的方法和介质产生的多肽; 和含有这种多肽的药物组合物。 合理设计的培养基含有计算用于细胞质量的氨基酸浓度，计算用于细胞维持的氨基酸浓度，以及计算用于掺入多肽的氨基酸浓度 出于兴趣。

公开(公告)号：US07294484B2

公开(公告)日：2007-11-13

申请号：US11213308

申请日：2005-08-25

申请人： Denis Drapeau ,  Yen-Tung Luan ,  James R. Mercer ,  Wenge Wang ,  Daniel R. Lasko

发明人： Denis Drapeau ,  Yen-Tung Luan ,  James R. Mercer ,  Wenge Wang ,  Daniel R. Lasko

IPC分类号： C12P21/06

CPC分类号： C12P21/02 ,  C07K16/18 ,  C07K16/22 ,  C07K16/2896 ,  C07K2317/24 ,  C12N5/0037 ,  C12N2500/22 ,  C12N2500/32 ,  C12N2500/38 ,  C12N2500/60 ,  C12N2510/02

摘要： An improved system for large scale production of proteins and/or polypeptides in cell culture, particularly in media characterized by one or more of: i) a cumulative amino acid concentration greater than about 70 mM; ii) a molar cumulative glutamine to cumulative asparagine ratio of less than about 2; iii) a molar cumulative glutamine to cumulative total amino acid ratio of less than about 0.2; iv) a molar cumulative inorganic ion to cumulative total amino acid ratio between about 0.4 to 1; or v) a combined cumulative glutamine and cumulative asparagine concentration between about 16 and 36 mM, is provided. The use of such a system allows high levels of protein production and lessens accumulation of certain undesirable factors such as ammonium and/or lactate. Additionally, culture methods including a temperature shift, typically including a decrease in temperature when the culture has reached about 20-80% of it maximal cell density, are provided. Alternatively or additionally, the present invention provides methods such that, after reaching a peak, lactate and/or ammonium levels in the culture decrease over time.

摘要翻译： 特别是在以下一个或多个特征的介质中大规模生产蛋白质和/或多肽的改进系统：i）累积的氨基酸浓度大于约70mM; ii）累积谷氨酰胺的累积天冬酰胺摩尔比小于约2; iii）累积谷氨酰胺与累积总氨基酸的摩尔比小于约0.2; iv）摩尔累计无机离子与累积总氨基酸比例在约0.4至1之间; 或v）组合的累积谷氨酰胺和约16至36mM之间的累积天冬酰胺浓度。 使用这样的系统允许高水平的蛋白质产生并减少某些不需要的因素例如铵和/或乳酸盐的积累。 此外，提供包括温度变化的培养方法，通常包括当培养物达到其最大细胞密度的约20-80％时的温度降低。 或者或另外，本发明提供了在达到峰值之后，培养物中的乳酸盐和/或铵水平随时间降低的方法。

公开(公告)号：US07300773B2

公开(公告)日：2007-11-27

申请号：US11213633

申请日：2005-08-25

申请人： Denis Drapeau ,  Yen-Tung Luan ,  James R. Mercer ,  Wenge Wang ,  Daniel R. Lasko

发明人： Denis Drapeau ,  Yen-Tung Luan ,  James R. Mercer ,  Wenge Wang ,  Daniel R. Lasko

IPC分类号： C12P21/06

CPC分类号： C12P21/02 ,  C07K14/7151 ,  C07K2319/30 ,  C12N2500/32 ,  C12N2500/90 ,  C12N2500/95 ,  C12P21/00

摘要： An improved system for large scale production of proteins and/or polypeptides in cell culture, particularly in media characterized by one or more of: i) a cumulative amino acid concentration greater than about 70 mM; ii) a molar cumulative glutamine to cumulative asparagine ratio of less than about 2; iii) a molar cumulative glutamine to cumulative total amino acid ratio of less than about 0.2; iv) a molar cumulative inorganic ion to cumulative total amino acid ratio between about 0.4 to 1; or v) a combined cumulative glutamine and cumulative asparagine concentration between about 16 and 36 mM, is provided. The use of such a system allows high levels of protein production and lessens accumulation of certain undesirable factors such as ammonium and/or lactate. Additionally, culture methods including a temperature shift, typically including a decrease in temperature when the culture has reached about 20-80% of it maximal cell density, are provided. Alternatively or additionally, the present invention provides methods such that, after reaching a peak, lactate and/or ammonium levels in the culture decrease over time.

摘要翻译： 特别是在以下一个或多个特征的介质中大规模生产蛋白质和/或多肽的改进系统：i）累积的氨基酸浓度大于约70mM; ii）累积谷氨酰胺的累积天冬酰胺摩尔比小于约2; iii）累积谷氨酰胺与累积总氨基酸的摩尔比小于约0.2; iv）摩尔累计无机离子与累积总氨基酸比例在约0.4至1之间; 或v）组合的累积谷氨酰胺和约16至36mM之间的累积天冬酰胺浓度。 使用这样的系统允许高水平的蛋白质产生并减少某些不需要的因素例如铵和/或乳酸盐的积累。 此外，提供包括温度变化的培养方法，通常包括当培养物达到其最大细胞密度的约20-80％时的温度降低。 或者或另外，本发明提供了在达到峰值之后，培养物中的乳酸盐和/或铵水平随时间降低的方法。

公开(公告)号：US20060121569A1

公开(公告)日：2006-06-08

申请号：US11213633

申请日：2005-08-25

申请人： Denis Drapeau ,  Yen-Tung Luan ,  James Mercer ,  Wenge Wang ,  Daniel Lasko

发明人： Denis Drapeau ,  Yen-Tung Luan ,  James Mercer ,  Wenge Wang ,  Daniel Lasko

IPC分类号： C12P21/06 ,  C12N5/06

CPC分类号： C12P21/02 ,  C07K14/7151 ,  C07K2319/30 ,  C12N2500/32 ,  C12N2500/90 ,  C12N2500/95 ,  C12P21/00

摘要： An improved system for large scale production of proteins and/or polypeptides in cell culture, particularly in media characterized by one or more of: i) a cumulative amino acid concentration greater than about 70 mM; ii) a molar cumulative glutamine to cumulative asparagine ratio of less than about 2; iii) a molar cumulative glutamine to cumulative total amino acid ratio of less than about 0.2; iv) a molar cumulative inorganic ion to cumulative total amino acid ratio between about 0.4 to 1; or v) a combined cumulative glutamine and cumulative asparagine concentration between about 16 and 36 mM, is provided. The use of such a system allows high levels of protein production and lessens accumulation of certain undesirable factors such as ammonium and/or lactate. Additionally, culture methods including a temperature shift, typically including a decrease in temperature when the culture has reached about 20-80% of it maximal cell density, are provided. Alternatively or additionally, the present invention provides methods such that, after reaching a peak, lactate and/or ammonium levels in the culture decrease over time.

摘要翻译： 特别是在以下一个或多个特征的介质中大规模生产蛋白质和/或多肽的改进系统：i）累积的氨基酸浓度大于约70mM; ii）累积谷氨酰胺的累积天冬酰胺摩尔比小于约2; iii）累积谷氨酰胺与累积总氨基酸的摩尔比小于约0.2; iv）摩尔累计无机离子与累积总氨基酸比例在约0.4至1之间; 或v）组合的累积谷氨酰胺和约16至36mM之间的累积天冬酰胺浓度。 使用这样的系统允许高水平的蛋白质产生并减少某些不需要的因素例如铵和/或乳酸盐的积累。 此外，提供包括温度变化的培养方法，通常包括当培养物达到其最大细胞密度的约20-80％时的温度降低。 或者或另外，本发明提供了在达到峰值之后，培养物中的乳酸盐和/或铵水平随时间降低的方法。

公开(公告)号：US08039231B2

公开(公告)日：2011-10-18

申请号：US12425056

申请日：2009-04-16

申请人： Yen-Tung Luan ,  Wenge Wang ,  Gregg Nyberg ,  Jose Manuel Gomes ,  Denis Drapeau ,  Terry Cardoza

发明人： Yen-Tung Luan ,  Wenge Wang ,  Gregg Nyberg ,  Jose Manuel Gomes ,  Denis Drapeau ,  Terry Cardoza

IPC分类号： C12P21/06

CPC分类号： C12P21/02 ,  A61K38/00 ,  C07K14/51 ,  C12N1/38 ,  C12N5/0037 ,  C12N2500/24 ,  C12N2500/38 ,  C12N2501/90 ,  C12N2510/02

摘要： Methods and processes for improved recombinant protein production are provided. The methods are useful for production of growth factors, particularly those of the TGF-β superfamily, including bone morphogenetic proteins (BMPs), such as BMP-2. Suitable host cells are cultured in media where iron is present at a concentration of at least 2.25 μM and if pyridoxal is present, it makes up less than 55% of the molar concentration of vitamin B6 in the media.

摘要翻译： 提供了改良重组蛋白生产的方法和方法。 该方法可用于生长生长因子，特别是TGF-β的生长因子; 超家族，包括骨形态发生蛋白（BMP），如BMP-2。 合适的宿主细胞在其中铁以至少2.25μM的浓度存在的培养基中培养，并且如果存在吡哆醛，则其占培养基中维生素​​B6的摩尔浓度的小于55％。

公开(公告)号：US20060160180A1

公开(公告)日：2006-07-20

申请号：US11213317

申请日：2005-08-25

申请人： Denis Drapeau ,  Yen-Tung Luan ,  James Mercer ,  Wenge Wang ,  Daniel Lasko

发明人： Denis Drapeau ,  Yen-Tung Luan ,  James Mercer ,  Wenge Wang ,  Daniel Lasko

IPC分类号： C12P21/06 ,  C12N5/06

CPC分类号： C07K16/28 ,  C07K16/00 ,  C07K16/18 ,  C07K16/22 ,  C07K2319/30 ,  C12N5/0018 ,  C12N2500/20 ,  C12N2500/32 ,  C12N2510/04 ,  C12P21/00

摘要： An improved system for large scale production of proteins and/or polypeptides in cell culture, particularly in media characterized by one or more of: i) a cumulative amino acid concentration greater than about 70 mM; ii) a molar cumulative glutamine to cumulative asparagine ratio of less than about 2; iii) a molar cumulative glutamine to cumulative total amino acid ratio of less than about 0.2; iv) a molar cumulative inorganic ion to cumulative total amino acid ratio between about 0.4 to 1; or v) a combined cumulative glutamine and cumulative asparagine concentration between about 16 and 36 mM, is provided. The use of such a system allows high levels of protein production and lessens accumulation of certain undesirable factors such as ammonium and/or lactate. Additionally, culture methods including a temperature shift, typically including a decrease in temperature when the culture has reached about 20-80% of it maximal cell density, are provided. Alternatively or additionally, the present invention provides methods such that, after reaching a peak, lactate and/or ammonium levels in the culture decrease over time.

摘要翻译： 特别是在以下一个或多个特征的介质中大规模生产蛋白质和/或多肽的改进系统：i）累积的氨基酸浓度大于约70mM; ii）累积谷氨酰胺的累积天冬酰胺摩尔比小于约2; iii）累积谷氨酰胺与累积总氨基酸的摩尔比小于约0.2; iv）摩尔累计无机离子与累积总氨基酸比例在约0.4至1之间; 或v）组合的累积谷氨酰胺和约16至36mM之间的累积天冬酰胺浓度。 使用这样的系统允许高水平的蛋白质产生并减少某些不需要的因素例如铵和/或乳酸盐的积累。 此外，提供包括温度变化的培养方法，通常包括当培养物达到其最大细胞密度的约20-80％时的温度降低。 或者或另外，本发明提供了在达到峰值之后，培养物中的乳酸盐和/或铵水平随时间降低的方法。

公开(公告)号：US20060121568A1

公开(公告)日：2006-06-08

申请号：US11213308

申请日：2005-08-25

申请人： Denis Drapeau ,  Yen-Tung Luan ,  James Mercer ,  Wenge Wang ,  Daniel Lasko

发明人： Denis Drapeau ,  Yen-Tung Luan ,  James Mercer ,  Wenge Wang ,  Daniel Lasko

IPC分类号： C12P21/06 ,  C07K14/705 ,  C12N5/06

CPC分类号： C12P21/02 ,  C07K16/18 ,  C07K16/22 ,  C07K16/2896 ,  C07K2317/24 ,  C12N5/0037 ,  C12N2500/22 ,  C12N2500/32 ,  C12N2500/38 ,  C12N2500/60 ,  C12N2510/02

摘要： An improved system for large scale production of proteins and/or polypeptides in cell culture, particularly in media characterized by one or more of: i) a cumulative amino acid concentration greater than about 70 mM; ii) a molar cumulative glutamine to cumulative asparagine ratio of less than about 2; iii) a molar cumulative glutamine to cumulative total amino acid ratio of less than about 0.2; iv) a molar cumulative inorganic ion to cumulative total amino acid ratio between about 0.4 to 1; or v) a combined cumulative glutamine and cumulative asparagine concentration between about 16 and 36 mM, is provided. The use of such a system allows high levels of protein production and lessens accumulation of certain undesirable factors such as ammonium and/or lactate. Additionally, culture methods including a temperature shift, typically including a decrease in temperature when the culture has reached about 20-80% of it maximal cell density, are provided. Alternatively or additionally, the present invention provides methods such that, after reaching a peak, lactate and/or ammonium levels in the culture decrease over time.

摘要翻译： 特别是在以下一个或多个特征的介质中大规模生产蛋白质和/或多肽的改进系统：i）累积的氨基酸浓度大于约70mM; ii）累积谷氨酰胺的累积天冬酰胺摩尔比小于约2; iii）累积谷氨酰胺与累积总氨基酸的摩尔比小于约0.2; iv）摩尔累计无机离子与累积总氨基酸比例在约0.4至1之间; 或v）组合的累积谷氨酰胺和约16至36mM之间的累积天冬酰胺浓度。 使用这样的系统允许高水平的蛋白质产生并减少某些不需要的因素例如铵和/或乳酸盐的积累。 此外，提供包括温度变化的培养方法，通常包括当培养物达到其最大细胞密度的约20-80％时的温度降低。 或者或另外，本发明提供了在达到峰值之后，培养物中的乳酸盐和/或铵水平随时间降低的方法。

公开(公告)号：US07335491B2

公开(公告)日：2008-02-26

申请号：US11213317

申请日：2005-08-25

申请人： Denis Drapeau ,  Yen-Tung Luan ,  James R. Mercer ,  Wenge Wang ,  Daniel R. Lasko

发明人： Denis Drapeau ,  Yen-Tung Luan ,  James R. Mercer ,  Wenge Wang ,  Daniel R. Lasko

IPC分类号： C12P21/06

CPC分类号： C07K16/28 ,  C07K16/00 ,  C07K16/18 ,  C07K16/22 ,  C07K2319/30 ,  C12N5/0018 ,  C12N2500/20 ,  C12N2500/32 ,  C12N2510/04 ,  C12P21/00

摘要： An improved system for large scale production of proteins and/or polypeptides in cell culture, particularly in media characterized by one or more of: i) a cumulative amino acid concentration greater than about 70 mM; ii) a molar cumulative glutamine to cumulative asparagine ratio of less than about 2; iii) a molar cumulative glutamine to cumulative total amino acid ratio of less than about 0.2; iv) a molar cumulative inorganic ion to cumulative total amino acid ratio between about 0.4 to 1; or v) a combined cumulative glutamine and cumulative asparagine concentration between about 16 and 36 mM, is provided. The use of such a system allows high levels of protein production and lessens accumulation of certain undesirable factors such as ammonium and/or lactate. Additionally, culture methods including a temperature shift, typically including a decrease in temperature when the culture has reached about 20-80% of it maximal cell density, are provided. Alternatively or additionally, the present invention provides methods such that, after reaching a peak, lactate and/or ammonium levels in the culture decrease over time.

摘要翻译： 特别是在以下一个或多个特征的介质中大规模生产蛋白质和/或多肽的改进系统：i）累积的氨基酸浓度大于约70mM; ii）累积谷氨酰胺的累积天冬酰胺摩尔比小于约2; iii）累积谷氨酰胺与累积总氨基酸的摩尔比小于约0.2; iv）摩尔累计无机离子与累积总氨基酸比例在约0.4至1之间; 或v）组合的累积谷氨酰胺和约16至36mM之间的累积天冬酰胺浓度。 使用这样的系统允许高水平的蛋白质产生并减少某些不需要的因素例如铵和/或乳酸盐的积累。 此外，提供包括温度变化的培养方法，通常包括当培养物达到其最大细胞密度的约20-80％时的温度降低。 或者或另外，本发明提供了在达到峰值之后，培养物中的乳酸盐和/或铵水平随时间降低的方法。