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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。
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公开(公告)号：US20070110743A1
公开(公告)日：2007-05-17
申请号：US11585607
申请日：2006-10-24
申请人： Denis Drapeau , Yen-Tung Luan , Paul Thoday , Wenge Wang , Judy Chou
发明人： Denis Drapeau , Yen-Tung Luan , Paul Thoday , Wenge Wang , Judy Chou
IPC分类号： A61K39/395 , C07H21/04 , C12P21/06 , C12N5/06 , C07K16/18
CPC分类号： C12N5/0018 , C12N2500/33 , C12N2501/998 , C12N2510/02 , C12P21/02
摘要： Methods of producing a protein in cell culture comprising an anti-senescence compound, such as the antioxidant carnosine, are provided. According to teachings of the present invention, cells grown in a cell culture medium comprising an anti-senescence compound exhibit increased viability and productivity. Furthermore, cell cultures grown in the presence of an anti-senescence compound exhibit decreased levels of high molecular weight aggregates in the cell culture medium.
摘要翻译： 提供了包含抗衰老化合物如抗氧化肌肽的细胞培养物中产生蛋白质的方法。 根据本发明的教导，在包含抗衰老化合物的细胞培养基中生长的细胞表现出增加的活力和生产力。 此外，在抗衰老化合物存在下生长的细胞培养物在细胞培养基中表现出降低的高分子量聚集体的水平。
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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。
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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
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。
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公开(公告)号：US08192951B2
公开(公告)日：2012-06-05
申请号：US11934458
申请日：2007-11-02
申请人： Wenge Wang , Yen Tung Luan
发明人： Wenge Wang , Yen Tung Luan
CPC分类号： C12N5/0018 , C12N2500/34 , C12N2510/02
摘要： An improved system for large scale production of proteins and/or polypeptides in cell culture is provided. In accordance with the present invention, cells expressing the protein or polypeptide of interest are grown in media that comprise a glycolysis-inhibiting substance. Additionally and/or alternatively, cells expressing the protein or polypeptide of interest are grown in media in which glutamine is limited. The use of such a system allows high levels of protein or polypeptide production and lessens accumulation of undesirable metabolic waste products such as lactate. Proteins and polypeptides expressed in accordance with the present invention may be advantageously used in the preparation of pharmaceutical, immunogenic, agricultural or other commercial compositions.
摘要翻译： 提供了在细胞培养物中大规模生产蛋白质和/或多肽的改进系统。 根据本发明，表达目的蛋白或多肽的细胞在包含糖酵解抑制物质的培养基中生长。 另外和/或替代地，表达目的蛋白质或多肽的细胞在限制谷氨酰胺的培养基中生长。 使用这种系统允许高水平的蛋白质或多肽产生并减少不合需要的代谢废物（例如乳酸盐）的积累。 根据本发明表达的蛋白质和多肽可有利地用于制备药物，免疫原性，农业或其它商业组合物。
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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％。
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公开(公告)号：US07913483B2
公开(公告)日：2011-03-29
申请号：US12371603
申请日：2009-02-15
申请人： Yafu Qiu , Weilin Xu , Dong Qiu , Cailing Ding , Weigang Cui , Wenge Wang , Xiaolin Shen , Aiguo Yang , Baocang Li , Jianping Wang , Guang Qin
发明人： Yafu Qiu , Weilin Xu , Dong Qiu , Cailing Ding , Weigang Cui , Wenge Wang , Xiaolin Shen , Aiguo Yang , Baocang Li , Jianping Wang , Guang Qin
IPC分类号： D02G3/36
摘要： On each draft element of a ring spinning frame, two pieces of short-staple roving from the roving bobbin enter into the draft mechanism to be drafted though a guide funnel in parallel, two pieces of filament are fed from the back of the front roller, and combine with two pieces of roving at front jaw respectively. The drafted two pieces of roving and filament are output from the front jaw and enter into the twisting triangle area to be twisted, and then are wound onto a yarn bobbin to produce yarn. Based on the relative position of the two pieces of roving and two pieces of filament, different yarns can be produced. Furthermore, multi-component yarn, such as core structure, wrapped structure, strand-like structure, can be produced on a ring spinning frame. The structure of yarn can be precisely determined. Special fiber yarn can be produced on traditional spinning frame.
摘要翻译： 在环锭纺纱机的每个牵引元件上，从粗纱筒管的两条短裤粗纱进入牵伸机构，通过平行的引导漏斗进行牵引，从前辊的后部进给两根细丝， 并分别与前颚两条粗纱相结合。 将两条粗纱和长丝从前颚输出并进入扭转三角形区域进行扭曲，然后缠绕在纱线架上以产生纱线。 基于两条粗纱和两条长丝的相对位置，可以生产不同的纱线。 此外，可以在环锭纺纱机上制造诸如芯结构，包裹结构，线状结构的多组分纱线。 可以精确确定纱线的结构。 特殊纤维纱可以在传统的纺纱机上生产。
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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
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。
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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
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。
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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。
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