公开(公告)号：US5643792A

公开(公告)日：1997-07-01

申请号：US181242

申请日：1994-01-13

申请人： Ken Okabayashi ,  Takao Ohmura ,  Kazumasa Yokoyama ,  Haruhide Kawabe

发明人： Ken Okabayashi ,  Takao Ohmura ,  Kazumasa Yokoyama ,  Haruhide Kawabe

IPC分类号： C12N1/20 ,  C07K14/765 ,  C12N1/15 ,  C12N1/21 ,  C12N9/04 ,  C12N9/64 ,  C12N15/00 ,  C12N15/01 ,  C12N15/52 ,  C12N15/74 ,  C12N15/78 ,  C12N15/81 ,  C12P21/00 ,  C12R1/84 ,  C12N15/11

CPC分类号： C12N9/0006 ,  C07K14/765 ,  C12N15/52 ,  C12N15/74 ,  C12N15/78 ,  C12N15/815 ,  C12Y101/03013

摘要： A methylotrophic and glucotrophic mutant strain capable of producing a heterologous protein and a method for producing a heterologous protein, comprising culture of the mutant strain. The mutant strain of the present invention can be grown in a medium containing both methanol and glucose, with the effect that the growth of the strain and production of a heterologous protein proceed at the same time. Accordingly, a heterologous protein can be produced in a large amount in a short time.

摘要翻译： 能够产生异源蛋白质的甲基营养型和葡萄糖营养型突变菌株和产生异源蛋白质的方法，包括突变菌株的培养。 本发明的突变菌株可以在含有甲醇和葡萄糖的培养基中生长，其效果是菌株的生长和异源蛋白质的产生同时进行。 因此，可以在短时间内大量生产异源蛋白质。

公开(公告)号：US5369020A

公开(公告)日：1994-11-29

申请号：US031823

申请日：1993-03-16

申请人： Akinori Sumi ,  Wataru Ohtani ,  Naoto Furuhata ,  Kazuya Takeshima ,  Kaeko Kamide ,  Takao Ohmura ,  Kazumasa Yokoyama

发明人： Akinori Sumi ,  Wataru Ohtani ,  Naoto Furuhata ,  Kazuya Takeshima ,  Kaeko Kamide ,  Takao Ohmura ,  Kazumasa Yokoyama

IPC分类号： A61K38/16 ,  A61P7/00 ,  B01D15/00 ,  C07K14/765 ,  C12N1/15 ,  C12N1/19 ,  C12N15/09 ,  C12N15/14 ,  C12P21/00 ,  C12P21/02 ,  C12R1/84 ,  C12R1/865 ,  C12P1/02

CPC分类号： C12P21/02 ,  C07K14/765

摘要： A method for suppressing coloring of human serum albumin expressed by genetic engineering, which comprises culture and/or purification in the presence of an amine compound selected from the group consisting of alkylamines, diamines, guanidines, benzamidines, basic amino acids, and aminophenylacetic acids. According to the present invention, coloring of HSA expressed by genetic engineering can be suppressed to from one-half to one-tenth of that without treatment for coloring suppression. In addition, HSA can be recovered in high yields, and the treatment of the invention does not affect the inherent properties of HSA.

摘要翻译： 一种用于抑制由遗传工程表达的人血清白蛋白着色的方法，其包括在选自烷基胺，二胺，胍胺，苯甲脒，碱性氨基酸和氨基苯基乙酸的胺化合物存在下培养和/或纯化。 根据本发明，通过遗传工程表达的HSA的着色可以抑制在不进行着色抑制处理的情况下的一半到十分之一。 另外，HSA可以以高收率回收，本发明的处理不影响HSA的固有特性。

公开(公告)号：US5440018A

公开(公告)日：1995-08-08

申请号：US36387

申请日：1993-03-24

申请人： Takao Ohmura ,  Akinori Sumi ,  Wataru Ohtani ,  Naoto Fuluhata ,  Kazuya Takeshima ,  Kaeko Kamide ,  Munehiro Noda ,  Masahide Kondo ,  Syoichi Ishikawa ,  Kazuhiro Oohara ,  Kazumasa Yokoyama

发明人： Takao Ohmura ,  Akinori Sumi ,  Wataru Ohtani ,  Naoto Fuluhata ,  Kazuya Takeshima ,  Kaeko Kamide ,  Munehiro Noda ,  Masahide Kondo ,  Syoichi Ishikawa ,  Kazuhiro Oohara ,  Kazumasa Yokoyama

IPC分类号： A61K38/00 ,  C07K1/14 ,  C07K14/765 ,  C12N15/14 ,  A61K38/38 ,  C07K1/16 ,  C07K1/36

CPC分类号： C07K14/765 ,  A61K38/00

摘要： Human serum albumin obtained by gene manipulation techniques can be purified by a combination of specified steps in which a culture supernatant obtained from a human serum albumin-producing host is subjected to ultrafiltration, heat treatment, acid treatment and another ultrafiltration, followed by subsequent treatments with a cation exchanger, a hydrophobic chromatography carrier and an anion exchanger, and by salting-out to thereby obtain a pure form of human serum albumin which contains substantially no proteinous and polysaccharide contaminants, which is formulated into a pharmaceutical preparation. This process makes it possible to effeciently purify recombinant human serum albumin and to provide substantially pure human serum albumin which does not contain producer host-related substances and other contaminants and is sufficiently free from coloration.

摘要翻译： 通过基因操作技术获得的人血清白蛋白可以通过特定步骤的组合来纯化，其中将得自人血清白蛋白的宿主的培养上清液进行超滤，热处理，酸处理和另一次超滤，然后用 阳离子交换剂，疏水性色谱载体和阴离子交换剂，并通过盐析，从而获得纯的形式的人血清白蛋白，其基本上不含有配制成药物制剂的蛋白质和多糖污染物。 该方法使得可以有效地纯化重组人血清白蛋白并提供基本上纯的人血清白蛋白，其不含生产者宿主相关物质和其它污染物，并且没有着色。

公开(公告)号：US5521287A

公开(公告)日：1996-05-28

申请号：US202130

申请日：1994-02-25

申请人： Takao Ohmura ,  Akinori Sumi ,  Wataru Ohtani ,  Naoto Furuhata ,  Kazuya Takeshima ,  Kaeko Kamide ,  Munehiro Noda ,  Masahide Kondo ,  Syoichi Ishikawa ,  Kazuhiro Oohara ,  Kazumasa Yokoyama ,  Nagatoshi Fujiwara

发明人： Takao Ohmura ,  Akinori Sumi ,  Wataru Ohtani ,  Naoto Furuhata ,  Kazuya Takeshima ,  Kaeko Kamide ,  Munehiro Noda ,  Masahide Kondo ,  Syoichi Ishikawa ,  Kazuhiro Oohara ,  Kazumasa Yokoyama ,  Nagatoshi Fujiwara

IPC分类号： A61K38/00 ,  C07K14/765 ,  A61K38/38 ,  C07K1/16 ,  C07K1/36

CPC分类号： C07K14/765 ,  A61K2039/545 ,  A61K38/00

摘要： Human serum albumin obtained by gene manipulation techniques can be purified by a combination of specified steps in which a culture supernatant obtained from a human serum albumin-producing host is subjected to ultrafiltration, heat treatment, acid treatment and another ultrafiltration, followed by subsequent treatments with a cation exchanger, a hydrophobic chromatography carrier and an anion exchanger, and by salting-out to thereby obtain a pure form of human serum albumin which contains substantially no proteinous and polysaccharide contaminants, which is formulated into a pharmaceutical preparation. The thus obtained human serum albumin can further be purified by treating recombinant human serum albumin with a hydrophobic chromatography carrier at pH of 2 to 5 and a salt concentration of 0.4 to 1 and exposing the carrier to a pH of 6 to 8 and a salt concentration of 0.01 to 0.3 M, or treating the culture supernatant with boric acid or a salt thereof at pH 8 to 11 for 1 to 10 hours and recovering the supernatant. This process makes it possible to effeciently purify recombinant human serum albumin and to provide substantially pure human serum albumin which does not contain producer host-related substances and other contaminants and is sufficiently free from coloration.

公开(公告)号：US5151499A

公开(公告)日：1992-09-29

申请号：US464077

申请日：1990-01-12

申请人： Shoju Kameyama ,  Kenmi Miyano ,  Motonori Hashimoto ,  Kazuo Takechi ,  Takao Ohmura ,  Yutaka Hirao ,  Yahiro Uemura ,  Kazumasa Yokoyama

发明人： Shoju Kameyama ,  Kenmi Miyano ,  Motonori Hashimoto ,  Kazuo Takechi ,  Takao Ohmura ,  Yutaka Hirao ,  Yahiro Uemura ,  Kazumasa Yokoyama

IPC分类号： A61K38/16 ,  A61K35/16 ,  A61K38/00 ,  A61K38/21 ,  A61K38/43 ,  A61K38/46 ,  A61K38/48 ,  A61K38/55 ,  A61K47/24 ,  A61L2/00 ,  C12N7/06

CPC分类号： C12N7/00 ,  A61K35/16 ,  A61L2/0023 ,  A61L2/0088 ,  C12N2760/20263

摘要： A method of producing a virus-inactivated protein-containing composition from a protein-containing composition which may be contaminated with virus. The method according to the present invention permits production of pharmaceutically safer virus-inactivated protein preparations without spoiling the protein activity.

公开(公告)号：US06617133B1

公开(公告)日：2003-09-09

申请号：US09317214

申请日：1999-05-24

申请人： Munehiro Noda ,  Akinori Sumi ,  Takao Ohmura ,  Kazumasa Yokoyama

发明人： Munehiro Noda ,  Akinori Sumi ,  Takao Ohmura ,  Kazumasa Yokoyama

IPC分类号： C12P2100

CPC分类号： C07K14/765

摘要： The invention provides a process for purifying recombinant human serum albumin (rHSA) by heating a-culture medium containing rHSA and the rHSA-producing host cells, feeding said heated solution upwardly into a fluidized bed in which adsorbent particles are suspended to effect contacting with the adsorbent particles and then recovering the adsorbed fraction containing the rHSA, and a composition comprising rHSA which shows a A350/A280 ratio of below 0.015, when formulated into a 25% solution of said albumin.

摘要翻译： 本发明提供了通过加热含有rHSA和产生rHSA的宿主细胞的培养基来纯化重组人血清白蛋白（rHSA）的方法，将所述加热溶液向上加入流化床中，其中吸附剂颗粒悬浮以实现与 吸附剂颗粒，然后回收含有rHSA的吸附级分，以及包含rHSA的组合物，其在配制成所述白蛋白的25％溶液时显示A350 / A280比率低于0.015。

公开(公告)号：US5962649A

公开(公告)日：1999-10-05

申请号：US522089

申请日：1995-08-31

申请人： Munehiro Noda ,  Akinori Sumi ,  Takao Ohmura ,  Kazumasa Yokoyama

发明人： Munehiro Noda ,  Akinori Sumi ,  Takao Ohmura ,  Kazumasa Yokoyama

IPC分类号： C07K14/765 ,  C07K1/16

CPC分类号： C07K14/765

摘要： The invention provides a process for purifying recombinant human serum albumin (rHSA) by heating a culture medium containing rHSA and the rHSA-producing host cello, feeding said heated solution upwardly into a fluidized bed in which adsorbent particles are suspended to effect contacting with the adsorbent particles and then recovering the adsorbed fraction containing the rHSA, and a composition comprising rHSA which shows a A35D/A280 ratio of below 0.015, when formulated into a 25% solution of said albumin.

摘要翻译： 本发明提供了通过加热含有rHSA和产生rHSA的宿主大肠杆菌的培养基来纯化重组人血清白蛋白（rHSA）的方法，将所述加热溶液向上加入到流化床中，其中吸附剂颗粒悬浮以与吸附剂接触 颗粒，然后回收含有rHSA的吸附级分，以及包含rHSA的组合物，其在配制成所述白蛋白的25％溶液时显示A35D / A280比率低于0.015。

公开(公告)号：US5986062A

公开(公告)日：1999-11-16

申请号：US538471

申请日：1995-10-03

申请人： Takao Ohmura ,  Akinori Sumi ,  Wataru Ohtani ,  Naoto Furuhata ,  Kazuya Takeshima ,  Kaeko Kamide ,  Munehiro Noda ,  Masahide Kondo ,  Syoichi Ishikawa ,  Kazuhiro Oohara ,  Kazumasa Yokoyama ,  Nagatoshi Fujiwara

发明人： Takao Ohmura ,  Akinori Sumi ,  Wataru Ohtani ,  Naoto Furuhata ,  Kazuya Takeshima ,  Kaeko Kamide ,  Munehiro Noda ,  Masahide Kondo ,  Syoichi Ishikawa ,  Kazuhiro Oohara ,  Kazumasa Yokoyama ,  Nagatoshi Fujiwara

IPC分类号： A61K38/00 ,  C07K14/765

CPC分类号： C07K14/765 ,  A61K2039/545 ,  A61K38/00

摘要： Human serum albumin obtained by gene manipulation techniques can be purified by a combination of specified steps in which a culture supernatant obtained from a human serum albumin-producing host is subjected to ultrafiltration, heat treatment, acid treatment and another ultrafiltration, followed by subsequent treatments with a cation exchanger, a hydrophobic chromatography carrier and an anion exchanger, and by salting-out to thereby obtain a pure form of human serum albumin which contains substantially no proteinous and polysaccharide contaminants, which is formulated into a pharmaceutical preparation. The thus obtained human serum albumin can further be purified by treating recombinant human serum albumin with a hydrophobic chromatography carrier at pH of 2 to 5 and a salt concentration of 0.4 to 1 and exposing the carrier to a pH of 6 to 8 and a salt concentration of 0.01 to 0.3 M, or treating the culture supernatant with boric acid or a salt thereof at pH 8 to 11 for 1 to 10 hours and recovering the supernatant. This process makes it possible to effeciently purify recombinant human serum albumin and to provide substantially pure human serum albumin which does not contain producer host-related substances and other contaminants and is sufficiently free from coloration.

摘要翻译： 通过基因操作技术获得的人血清白蛋白可以通过特定步骤的组合来纯化，其中将得自人血清白蛋白的宿主的培养上清液进行超滤，热处理，酸处理和另一次超滤，然后用 阳离子交换剂，疏水性色谱载体和阴离子交换剂，并通过盐析，从而获得纯的形式的人血清白蛋白，其基本上不含有配制成药物制剂的蛋白质和多糖污染物。 由此获得的人血清白蛋白可以进一步通过用pH为2至5和盐浓度为0.4至1的疏水色谱载体处理重组人血清白蛋白并将载体暴露于pH6至8和盐浓度 为0.01〜0.3M，或用硼酸或其盐在pH8〜11处理培养上清1〜10小时，回收上清液。 该方法使得可以有效地纯化重组人血清白蛋白并提供基本上纯的人血清白蛋白，其不含生产者宿主相关物质和其它污染物，并且没有着色。

公开(公告)号：US5334512A

公开(公告)日：1994-08-02

申请号：US854841

申请日：1992-03-20

申请人： Kaoru Kobayashi ,  Shinobu Kuwae ,  Tomoshi Ooya ,  Hirotoshi Fukutsuka ,  Akinori Sumi ,  Wataru Ohtani ,  Takao Ohmura ,  Kazumasa Yokoyama

发明人： Kaoru Kobayashi ,  Shinobu Kuwae ,  Tomoshi Ooya ,  Hirotoshi Fukutsuka ,  Akinori Sumi ,  Wataru Ohtani ,  Takao Ohmura ,  Kazumasa Yokoyama

IPC分类号： C07K14/765 ,  C12N1/16 ,  C12N1/19 ,  C12N15/09 ,  C12N15/14 ,  C12P21/00 ,  C12R1/84 ,  C12N1/38 ,  C12N15/81 ,  C12P21/02

CPC分类号： C07K14/765 ,  C12N1/16

摘要： A method for producing human serum albumin which comprises cultivating a human serum albumin-producing host prepared by genetic engineering, in a medium containing a fatty acid having 10 to 26 carbon atoms, or its salt, and a method for cultivating the host. HSA production can be greatly increased by the present invention.

摘要翻译： 一种生产人血清白蛋白的方法，包括在含有10-26个碳原子的脂肪酸的培养基或其盐中培养通过基因工程制备的人血清白蛋白生产宿主及其培养方法。 通过本发明可以大大增加HSA的生产。

公开(公告)号：US5683893A

公开(公告)日：1997-11-04

申请号：US471206

申请日：1995-06-06

申请人： Hideyuki Ohi ,  Masami Miura ,  Shusei Uno ,  Masako Chuganji ,  Ryuji Hiramatsu ,  Takao Ohmura

发明人： Hideyuki Ohi ,  Masami Miura ,  Shusei Uno ,  Masako Chuganji ,  Ryuji Hiramatsu ,  Takao Ohmura

IPC分类号： C07K14/765 ,  C12N1/19 ,  C12N15/09 ,  C12N15/14 ,  C12N15/81 ,  C12P21/02 ,  C12R1/84 ,  C12P21/00 ,  C07H21/04 ,  C12N15/64

CPC分类号： C12N15/815 ,  C07K14/765

摘要： A mutant AOX2 promoter obtained by mutating a sequence of natural AOX2 promoter in a manner comprising at least one of the three mutation modes of (1) a region extending upstream from nucleotide 1187 inclusive and comprising at least nucleotides 845-960 is deleted, (2) nucleotide(s) is(are) replaced in region(s) in nucleotides 1274-1314, and (3) new oligonucleotide(s) is (are) inserted in region(s) in nucleotides 1274-1314, a vector carrying said mutant AOX2 promoter, a transformant into which said vector has been introduced, and a method for producing a heterologous protein, which comprises cultivating said transformant. The promoter of the present invention has remarkably enhanced activity as compared with natural AOX2 promoter, and is highly useful as a promoter to be carried in an expression vector allowing heterologous protein expression. In addition, the vector and the transformant of the invention can efficiently express and produce various useful heterologous proteins.

摘要翻译： 以包含以下三种突变模式中的至少一种的方式突变天然AOX2启动子的突变体AOX2启动子被缺失：（1）包含第1187位核苷酸并且至少包含核苷酸845-960的上游延伸的区域;（2） ）核苷酸在核苷酸1274-1314的区域中被替换，并且（3）将新的寡核苷酸插入到核苷酸1274-1314的区域中，载体携带所述 突变体AOX2启动子，其中引入了所述载体的转化体，以及产生异源蛋白质的方法，其包括培养所述转化体。 与天然AOX2启动子相比，本发明的启动子具有显着增强的活性，并且作为允许异源蛋白质表达的表达载体中携带的启动子是非常有用的。 此外，本发明的载体和转化体可以有效地表达和产生各种有用的异源蛋白质。