公开(公告)号：US20070059794A1

公开(公告)日：2007-03-15

申请号：US10554747

申请日：2004-04-28

申请人： Akira Ideno ,  Jun-ichi Hata ,  Akiko Togi ,  Masahiro Furutani

发明人： Akira Ideno ,  Jun-ichi Hata ,  Akiko Togi ,  Masahiro Furutani

IPC分类号： C12P21/06 ,  C07H21/04 ,  C12N9/64

CPC分类号： C12N15/62 ,  A61K38/1709 ,  C07K2319/20 ,  C07K2319/35 ,  C07K2319/40 ,  C07K2319/705 ,  G01N33/6803

摘要： The present invention provides a chaperonin-target protein complex and a method of producing the same, and a method of stabilizing the target protein, a method of immobilizing the target protein, a method of analyzing the structure of the target protein, a sustained-release formulation, and a method of producing an antibody against the target protein. The chaperonin-target protein complex in the present invention includes a fusion protein having a chaperonin subunit and an affinity tag linked to the chaperonin subunit via a peptide bond and a target protein for which the affinity tag shows a specific affinity, wherein the target protein is bound to the affinity tag by means of the specific affinity, thereby forming a chaperonin ring structure consisting of a plurality of chaperonin subunits. The chaperonin-target protein in the present invention stabilizes the target protein and surely immobilize on a carrier without causing any change in its stereostructure.

摘要翻译： 本发明提供了伴侣伴侣 - 靶蛋白复合物及其制造方法，使靶蛋白稳定化的方法，固定靶蛋白的方法，分析靶蛋白的结构的方法，持续释放 制剂和产生针对靶蛋白的抗体的方法。 本发明中的伴侣伴侣 - 靶蛋白复合物包括伴侣伴侣亚基和通过肽键与伴侣蛋白亚基连接的亲和标签的融合蛋白和亲和标签显示特异性亲和力的靶蛋白，其中靶蛋白为 通过特异性亲和力结合亲和标签，从而形成由多个伴侣蛋白亚基组成的伴侣蛋白环结构。 本发明中的伴侣伴侣靶蛋白稳定靶蛋白，并确定地固定在载体上，而不引起其立体结构的任何改变。

公开(公告)号：US07276355B2

公开(公告)日：2007-10-02

申请号：US10451883

申请日：2001-12-26

申请人： Masahiro Furutani ,  Junichi Hata ,  Akiko Togi

发明人： Masahiro Furutani ,  Junichi Hata ,  Akiko Togi

IPC分类号： C12P21/04

CPC分类号： C12N15/62 ,  C07K19/00 ,  C07K2319/21 ,  C07K2319/35 ,  C07K2319/40 ,  C07K2319/75 ,  C12N15/70 ,  C12P21/02

摘要： This invention provides a recombinant protein expression system using a host and cell-free translation system, and is capable of universally expressing a large amount of any protein as soluble protein, while preventing toxicity in hosts, formation of inclusion bodies, and decomposition with proteases. Such may be achieved by expressing the desired protein as a fusion protein with chaperonins, such as about 60 kDa molecular chaperones, 60 kDa heat shock proteins, or thermosomes, and accommodating the desired protein inside of a stereostructure of a chaperonin. The present invention provides a process for producing a protein, which comprises transcribing and translating a gene containing a gene encoding the linked chaperonin subunits and a gene encoding a desired protein thereby synthesizing a fusion protein having the desired protein linked via a peptide linkage to the linked chaperonin subunits.

摘要翻译： 本发明提供了使用宿主和无细胞翻译系统的重组蛋白质表达系统，并且能够普遍表达大量的任何蛋白质作为可溶性蛋白质，同时防止宿主中的毒性，包涵体的形成和蛋白酶的分解。 这可以通过用伴侣蛋白（例如约60kDa的分子伴侣，60kDa的热休克蛋白或热单体）表达所需的蛋白质作为融合蛋白，并且将伴侣蛋白的立体结构中的所需蛋白质容纳。 本发明提供了一种生产蛋白质的方法，其包括转录和翻译含有编码连接的伴侣蛋白亚基的基因的基因和编码所需蛋白质的基因，从而合成具有通过肽键连接的所需蛋白质的融合蛋白与所连接的 伴侣亚单位。

公开(公告)号：US20070092937A1

公开(公告)日：2007-04-26

申请号：US10554751

申请日：2004-04-28

申请人： Akiko Togi ,  Masahiro Furutani

发明人： Akiko Togi ,  Masahiro Furutani

IPC分类号： C07K14/195 ,  C12P21/06 ,  C07H21/04 ,  C12N15/74 ,  C12N1/21 ,  C12N1/18

CPC分类号： C12P21/06 ,  C07K2319/23 ,  C07K2319/35 ,  C07K2319/43 ,  C07K2319/61 ,  C07K2319/92 ,  C12N15/62

摘要： It is intended to provide techniques whereby a target protein is efficiently obtained as a normal protein even in the case of a protein which can be hardly expressed by usual recombinant DNA techniques. A fusion protein composed of a molecular chaperone, an intervening protein having an activity of cleaving a peptide bond, and a target protein is prepared, from which fusion protein the target protein is cleaved by the action of the intervening protein having the activity of cleaving a peptide bond. Examples of the intervening protein having the activity of cleaving a peptide bond include an intein and a protein consisting of a partial sequence of the intein.

摘要翻译： 旨在提供即使在通常的重组DNA技术几乎不能表达的蛋白质的情况下也可有效地获得靶蛋白作为正常蛋白质的技术。 制备由分子伴侣组成的融合蛋白，具有切割肽键的活性的中间蛋白质和靶蛋白质，通过具有切割活性的中间蛋白质的作用将靶蛋白切割成融合蛋白 肽键。 具有切割肽键活性的中间蛋白质的实例包括内含肽和由内含肽的部分序列组成的蛋白质。

公开(公告)号：US07608424B2

公开(公告)日：2009-10-27

申请号：US11837795

申请日：2007-08-13

申请人： Masahiro Furutani ,  Junichi Hata ,  Akiko Togi

发明人： Masahiro Furutani ,  Junichi Hata ,  Akiko Togi

IPC分类号： C12P21/04

CPC分类号： C12N15/62 ,  C07K19/00 ,  C07K2319/21 ,  C07K2319/35 ,  C07K2319/40 ,  C07K2319/75 ,  C12N15/70 ,  C12P21/02

摘要： This invention provides a recombinant protein expression system using a host and cell-free translation system, and is capable of universally expressing a large amount of any protein as soluble protein, while preventing toxicity in hosts, formation of inclusion bodies, and decompositions with proteases. Such may be achieved by expressing the desired protein as a fusion protein with chaperoning, such as about 60 kDa molecular chaperons, 60 kDa heat shock proteins, or thermosomes, and accommodating the desired protein inside of a stereostructure of a chaperonin. The present invention provides a process for producing a protein, which comprises transcribing and translating a gene containing a gene encoding the linked chaperonin subunits and a gene encoding a desired protein thereby synthesizing a fusion protein having the desired protein linked via a peptide linkage to the linked chaperonin subunits.

摘要翻译： 本发明提供了使用宿主和无细胞翻译系统的重组蛋白质表达系统，并且能够普遍表达大量的任何蛋白质作为可溶性蛋白质，同时防止宿主中的毒性，包涵体的形成和蛋白酶的分解。 这可以通过用伴侣蛋白表达所需蛋白质作为融合蛋白，例如约60kDa的分子伴侣，60kDa的热休克蛋白或者热单体，并且在伴侣蛋白的立体结构内容纳所需的蛋白质来实现。 本发明提供了一种生产蛋白质的方法，其包括转录和翻译含有编码连接的伴侣蛋白亚基的基因的基因和编码所需蛋白质的基因，从而合成具有通过肽键连接的所需蛋白质的融合蛋白与所连接的 伴侣亚单位。

公开(公告)号：US20080081361A1

公开(公告)日：2008-04-03

申请号：US11837795

申请日：2007-08-13

申请人： Masahiro FURUTANI ,  Junichi Hata ,  Akiko Togi

发明人： Masahiro FURUTANI ,  Junichi Hata ,  Akiko Togi

IPC分类号： C12N1/20 ,  C07K14/00 ,  C12N15/00

CPC分类号： C12N15/62 ,  C07K19/00 ,  C07K2319/21 ,  C07K2319/35 ,  C07K2319/40 ,  C07K2319/75 ,  C12N15/70 ,  C12P21/02

摘要： This invention provides a recombinant protein expression system using a host and cell-free translation system, and is capable of universally expressing a large amount of any protein as soluble protein, while preventing toxicity in hosts, formation of inclusion bodies, and decompositions with proteases. Such may be achieved by expressing the desired protein as a fusion protein with chaperoning, such as about 60 kDa molecular chaperons, 60 kDa heat shock proteins, or thermosomes, and accommodating the desired protein inside of a stereostructure of a chaperonin. The present invention provides a process for producing a protein, which comprises transcribing and translating a gene containing a gene encoding the linked chaperonin subunits and a gene encoding a desired protein thereby synthesizing a fusion protein having the desired protein linked via a peptide linkage to the linked chaperonin subunits.

摘要翻译： 本发明提供了使用宿主和无细胞翻译系统的重组蛋白质表达系统，并且能够普遍表达大量的任何蛋白质作为可溶性蛋白质，同时防止宿主中的毒性，包涵体的形成和蛋白酶的分解。 这可以通过用伴侣蛋白表达所需蛋白质作为融合蛋白，例如约60kDa的分子伴侣，60kDa的热休克蛋白或者热单体，并且在伴侣蛋白的立体结构内容纳所需的蛋白质来实现。 本发明提供了一种生产蛋白质的方法，其包括转录和翻译含有编码连接的伴侣蛋白亚基的基因的基因和编码所需蛋白质的基因，从而合成具有通过肽键连接的所需蛋白质的融合蛋白与所连接的 伴侣亚单位。

公开(公告)号：US20050260710A1

公开(公告)日：2005-11-24

申请号：US11092988

申请日：2005-03-30

申请人： Kazuo Suzuki ,  Masahiro Furutani ,  Kenji Yamamoto ,  Naohito Ohno ,  Kei Takahashi ,  Yasuaki Aratani ,  Akiko Togi

发明人： Kazuo Suzuki ,  Masahiro Furutani ,  Kenji Yamamoto ,  Naohito Ohno ,  Kei Takahashi ,  Yasuaki Aratani ,  Akiko Togi

IPC分类号： C07K16/00 ,  C07K16/44 ,  C12N1/21 ,  C12N15/74 ,  C12P21/06

CPC分类号： C07K16/00 ,  A61K2039/505 ,  C07K2317/52 ,  C07K2317/622 ,  C07K2319/00

摘要： The purpose of the present invention is to provide methods for producing recombinant polyclonal immunoglobulins using recombinant DNA techniques, which provides constant supply of immunoglobulin preparations with minimum risk of infection. A method of the present invention comprises the following steps: (1) isolating a plurality of types of genes from cDNAs derived from tissues or cells expressing immunoglobulins, said genes encoding a plurality of types of polypeptides respectively, and said polypeptides containing a plurality of types of immunoglobulin variable regions respectively, and preparing mixture of the genes; (2) preparing a plurality of types of recombinant vectors into which a plurality of types of genes are introduced respectively by contacting said mixture of the genes with vectors, and preparing mixture of the recombinant vectors; (3) preparing a plurality of types of transformants into which a plurality of types of recombinant vectors are introduced respectively by contacting said mixture of the recombinant vectors with host cells, and preparing mixture of the transformants; and (4) culturing said mixture of the transformants, and obtaining mixture of polypeptides from the transformants culture, wherein said polypeptides contain a plurality of types of immunoglobulin variable regions respectively.

摘要翻译： 本发明的目的是提供使用重组DNA技术制备重组多克隆免疫球蛋白的方法，其提供具有最小感染风险的免疫球蛋白制剂的恒定供应。 本发明的方法包括以下步骤：（1）从衍生自表达免疫球蛋白的组织或细胞的cDNA分离多种类型的基因，所述基因分别编码多种类型的多肽，所述多肽含有多种类型 的免疫球蛋白可变区，并制备基因的混合物; （2）通过将所述基因混合物与载体接触，制备多种类型的重组载体，其中分别导入多种类型的基因，并制备重组载体的混合物; （3）通过使重组载体的混合物与宿主细胞接触，制备分别引入多种类型的重组载体的多种类型的转化体，并制备转化体的混合物; （4）培养所述转化体的所述混合物，并从所述转化体培养物获得多肽的混合物，其中所述多肽分别含有多种类型的免疫球蛋白可变区。