公开(公告)号：US20090299035A1

公开(公告)日：2009-12-03

申请号：US12443623

申请日：2007-10-10

申请人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota ,  Gou Sarara ,  Yukiko Aruga ,  Chiori Yamane

发明人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota ,  Gou Sarara ,  Yukiko Aruga ,  Chiori Yamane

IPC分类号： C07K14/00

CPC分类号： C07K14/31 ,  C07K14/315 ,  C07K17/06 ,  C07K2319/21

摘要： An object of the present invention is to provide a novel protein having the following amino acid sequence altered for specifically and efficiently binding a protein to an immobilization carrier via the carboxy terminus. The protein is used for immobilizing a portion represented by R1-R2 on an immobilization carrier, comprising the amino acid sequence represented by the general formula R1-R2-R3-R4-R5 [wherein: the sequences are oriented from the amino terminal side to the carboxy terminal side; the sequence of the R1 portion is the sequence of a subject protein to be immobilized and contains neither a lysine residue nor a cysteine residue; the sequence of the R2 portion may be absent, but when the sequence of the R2 portion is present, the sequence of the R2 portion is a spacer sequence composed of amino acid residues other than lysine and cysteine residues; the sequence of the R3 portion is composed of two residues of amino acid represented by cysteine-X (where X denotes an amino acid residue other than lysine or cysteine); the sequence of the R4 portion may be absent, but when the sequence of the R4 portion is present, the sequence of the R4 portion contains neither a lysine residue nor a cysteine residue, but contains an acidic amino acid residue capable of acidifying the isoelectric point of the entire protein comprising the amino acid sequence represented by the general formula R1-R2-R3-R4-R5; and the sequence of an R5 portion is an affinity tag sequence for protein purification.

摘要翻译： 本发明的一个目的是提供一种具有以下氨基酸序列的新蛋白质，所述氨基酸序列经由羧基末端将蛋白质特异性和有效地结合到固定化载体上。 该蛋白质用于将由R1-R2表示的部分固定在固定化载体上，其包含由通式R1-R2-R3-R4-R5表示的氨基酸序列[其中：序列从氨基末端侧取向至 羧基末端侧; R1部分的序列是待固定的主题蛋白的序列，既不含有赖氨酸残基也不含有半胱氨酸残基; R2部分的序列可以不存在，但是当存在R2部分的序列时，R2部分的序列是由赖氨酸和半胱氨酸残基以外的氨基酸残基组成的间隔序列; R3部分的序列由半胱氨酸-X（其中X表示除赖氨酸或半胱氨酸以外的氨基酸残基）表示的两个氨基酸残基组成; R4部分的序列可以不存在，但当存在R4部分的序列时，R4部分的序列既不含有赖氨酸残基也不含有半胱氨酸残基，但含有能够酸化等电点的酸性氨基酸残基 包含由通式R1-R2-R3-R4-R5表示的氨基酸序列的整个蛋白质; 并且R5部分的序列是用于蛋白质纯化的亲和标签序列。

公开(公告)号：US20100130721A1

公开(公告)日：2010-05-27

申请号：US12596600

申请日：2008-04-17

申请人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota ,  Gou Sarara ,  Hisashi Takahashi ,  Yukiko Aruga ,  Chiori Yamane

发明人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota ,  Gou Sarara ,  Hisashi Takahashi ,  Yukiko Aruga ,  Chiori Yamane

IPC分类号： C07K17/00

CPC分类号： C07K17/00

摘要： This invention provides an immobilized protein bound to an immobilization carrier at a protein amino terminus via the sole α-amino group of the protein comprising an amino acid sequence containing neither lysine residues nor cysteine residues represented by the general formula S1-R1-R2, wherein: the sequences are oriented from the amino terminal side to the carboxy terminal side; the sequence of the S1 portion may be absent, but when the sequence of the S1 portion is present, the sequence of the S1 portion is a spacer sequence composed of amino acid residues other than lysine and cysteine residues; the sequence of the R1 portion is the sequence of a subject protein to be immobilized and contains neither lysine residues nor cysteine residues; and the sequence of the R2 portion may be absent, but when the sequence of the R2 portion is present, the sequence of the R2 portion is a spacer sequence composed of amino acid residues other than lysine and cysteine residues.

摘要翻译： 本发明提供了通过蛋白质的唯一α-氨基在蛋白质氨基末端与固定化载体结合的固定化蛋白，其包含不含有由通式S1-R1-R2表示的赖氨酸残基和半胱氨酸残基的氨基酸序列，其中 ：序列从氨基末端侧到羧基末端侧; S1部分的序列可以不存在，但是当存在S1部分的序列时，S1部分的序列是由赖氨酸和半胱氨酸残基以外的氨基酸残基组成的间隔序列; R1部分的序列是待固定的主题蛋白的序列，既不含有赖氨酸残基也不含有半胱氨酸残基; 并且R2部分的序列可以不存在，但是当R2部分的序列存在时，R2部分的序列是由赖氨酸和半胱氨酸残基以外的氨基酸残基组成的间隔序列。

公开(公告)号：US20080051555A1

公开(公告)日：2008-02-28

申请号：US10575254

申请日：2004-10-07

申请人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota

发明人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hiroyuki Sota

IPC分类号： C07K1/14 ,  C07K14/00

CPC分类号： C07K17/06

摘要： A support having an affinity for an antibody characterized in that the carboxy end of a protein or a peptide, which is capable of binding to an antibody molecule, is immobilized via an amide bond mediated by a linker sequence to an insoluble support having a primary amino group. This support has an excellent ability to adsorb antibody molecules.

摘要翻译： 具有对抗体亲和力的载体，其特征在于能够结合抗体分子的蛋白质或肽的羧基末端通过由接头序列介导的酰胺键固定到具有伯氨基的不溶性载体上 组。 该载体具有优异的吸附抗体分子的能力。

公开(公告)号：US20080146459A1

公开(公告)日：2008-06-19

申请号：US10553626

申请日：2004-04-09

申请人： Masahiro Iwakura ,  Kiyonori Hirota

发明人： Masahiro Iwakura ,  Kiyonori Hirota

IPC分类号： C40B40/10 ,  C40B50/18

CPC分类号： G01N33/6845 ,  B01J19/0046 ,  B01J2219/00497 ,  B01J2219/00527 ,  B01J2219/00585 ,  B01J2219/00596 ,  B01J2219/00605 ,  B01J2219/00659 ,  B01J2219/00725 ,  C07K17/02 ,  C40B40/10

摘要： A protein array having a protein aligned and immobilized in high density and in uniformly oriented form is produced by introducing a polymer compound having a primary amino group as a repeating unit on the surface of a planar substrate so as to obtain a substrate for protein array and binding the carboxyl terminal of the main chain of a protein with the primary amino group of the polymer compound on the substrate.

摘要翻译： 通过将具有作为重复单元的伯氨基作为重复单元的高分子化合物引入到平面基板的表面上，从而获得蛋白质排列的基板，从而制造具有以高密度和均匀定向形式排列并固定的蛋白质的蛋白质阵列 将蛋白质主链的羧基末端与高分子化合物的伯氨基结合在基材上。

公开(公告)号：US20080261827A1

公开(公告)日：2008-10-23

申请号：US11792095

申请日：2005-12-02

申请人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hisashi Takahashi

发明人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hisashi Takahashi

IPC分类号： C40B30/10 ,  C40B60/12 ,  C40B40/10

CPC分类号： G01N33/5308 ,  G01N21/253 ,  G01N21/31 ,  G01N21/6452 ,  G01N33/54386

摘要： To provide a method for detecting a protein by immobilizing a protein on a protein array substrate at a high density with controlled orientation, irradiating the immobilized protein with ultraviolet light, visible light, or infrared light, and measuring the light not absorbed by the protein and further analyzing an interaction between the protein on the substrate and another protein and/or a compound other than proteins, a system used for the method, and to provide a protein array suitable for the system. A system including a protein array in which a protein is immobilized in aligned position on a light-transmissive substrate at a high density, a light-irradiating means, and a light-detecting means and being for detecting or analyzing a protein on the protein array and/or a compound which is other than proteins and interacts with the immobilized protein, wherein the protein array is irradiated with light by the light-irradiating means, and the light transmitted through the protein array is measured by the light-detecting means for detecting the light absorption of the protein on the protein array and/or the compound which is other than proteins and interacts with the immobilized protein.

摘要翻译： 为了提供一种通过将具有受控取向的高密度蛋白质固定在蛋白质阵列基底上的蛋白质的检测方法，用紫外光，可见光或红外光照射固定化蛋白质，并测定不被蛋白质吸收的光， 进一步分析底物上的蛋白质与另一种蛋白质和/或蛋白质以外的化合物之间的相互作用，用于该方法的系统，以及提供适用于该系统的蛋白质阵列。 一种包含蛋白质阵列的系统，其中蛋白质以高密度固定在透光基底上的对准位置，光照射装置和光检测装置，并用于检测或分析蛋白质阵列上的蛋白质 和/或不同于蛋白质并与固定化蛋白质相互作用的化合物，其中通过光照射装置用光照射蛋白质阵列，通过蛋白质阵列传输的光由用于检测的光检测装置测量 蛋白质在蛋白质阵列上的光吸收和/或不同于蛋白质并与固定化蛋白质相互作用的化合物。

公开(公告)号：US08038945B2

公开(公告)日：2011-10-18

申请号：US11792095

申请日：2005-12-02

申请人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hisashi Takahashi

发明人： Masahiro Iwakura ,  Kiyonori Hirota ,  Hisashi Takahashi

IPC分类号： G01N21/00

CPC分类号： G01N33/5308 ,  G01N21/253 ,  G01N21/31 ,  G01N21/6452 ,  G01N33/54386

摘要： To provide a method for detecting a protein by immobilizing a protein on a protein array substrate at a high density with controlled orientation, irradiating the immobilized protein with ultraviolet light, visible light, or infrared light, and measuring the light not absorbed by the protein and further analyzing an interaction between the protein on the substrate and another protein and/or a compound other than proteins, a system used for the method, and to provide a protein array suitable for the system. A system including a protein array in which a protein is immobilized in aligned position on a light-transmissive substrate at a high density, a light-irradiating means, and a light-detecting means and being for detecting or analyzing a protein on the protein array and/or a compound which is other than proteins and interacts with the immobilized protein, wherein the protein array is irradiated with light by the light-irradiating means, and the light transmitted through the protein array is measured by the light-detecting means for detecting the light absorption of the protein on the protein array and/or the compound which is other than proteins and interacts with the immobilized protein.

摘要翻译： 为了提供一种通过将具有受控取向的高密度蛋白质固定在蛋白质阵列基底上的蛋白质的检测方法，用紫外光，可见光或红外光照射固定化蛋白质，并测定不被蛋白质吸收的光， 进一步分析底物上的蛋白质与另一种蛋白质和/或蛋白质以外的化合物之间的相互作用，用于该方法的系统，以及提供适用于该系统的蛋白质阵列。 一种包含蛋白质阵列的系统，其中蛋白质以高密度固定在透光基底上的对准位置，光照射装置和光检测装置，并用于检测或分析蛋白质阵列上的蛋白质 和/或不同于蛋白质并与固定化蛋白质相互作用的化合物，其中通过光照射装置用光照射蛋白质阵列，通过蛋白质阵列传输的光由用于检测的光检测装置测量 蛋白质在蛋白质阵列上的光吸收和/或不同于蛋白质并与固定化蛋白质相互作用的化合物。

公开(公告)号：US20090011958A1

公开(公告)日：2009-01-08

申请号：US11758629

申请日：2007-06-05

申请人： Masahiro Iwakura ,  Kiyonori Hirota

发明人： Masahiro Iwakura ,  Kiyonori Hirota

IPC分类号： C40B50/06 ,  C07K17/02 ,  C12N11/02

CPC分类号： C07K17/06 ,  C07K14/001

摘要： This invention provides a process for effectively producing an immobilized protein that is immobilized only at the carboxy terminus. This is a process for immobilizing on a carrier for immobilization a protein represented by general formula (1): NH2—R1—COOH   (1) wherein a protein having a sulfhydryl group represented by general formula (2) is produced: NH2—R1—CONH—R2—CO—NH—CH(CH2—SH)—CO—NH—R3—COOH   (2), the resulting protein is immobilized adsorptively on the carrier for immobilization represented by general formula (3) via ion interactions under neutral conditions: NH2—Y   (3), and a sulfhydryl group of the cysteine residue in the protein represented by general formula (2) that is adsorbed on the carrier for immobilization represented by general formula (3) is cyanated using a cyanating reagent to convert it into a cyanocysteine residue,thereby producing an immobilized protein represented by general formula (4): NH2—R1—CO—NH—R2—CO—NH—Y   (4) wherein R1 and R2 represent arbitrary amino acid sequences, R3 represents an amino acid sequence that has a strong negative charge around the neutral region and can acidify the isoelectric point of NH2—R1—CONH—R2—CO—NH—CH(CH2—SH)—CO—NH—R3—COOH, and Y represents an arbitrary carrier for immobilization.

公开(公告)号：US20070015241A1

公开(公告)日：2007-01-18

申请号：US10959021

申请日：2004-10-04

申请人： Masahiro Iwakura

发明人： Masahiro Iwakura

IPC分类号： C12N9/24 ,  C07H21/04 ,  C12P21/06 ,  C12N15/74 ,  C12N1/21

CPC分类号： C12N9/003 ,  C07K2319/00 ,  C12N9/00 ,  C12N9/0065 ,  C12N9/248 ,  C12Y302/01008

摘要： A sulfur atom-free enzyme protein retaining the activity of the original enzyme protein and having oxidation resistance wherein L-cystein and L-methionine residues of the enzyme protein are substituted with 18 types of L-amino acid residues: L-alanine; L-aspartic acid; L-glutamic acid; L-phenylalanine; L-glycine; L-histidine; L-isoleucine; L-lysine; L-leucine; L-asparagine; L-proline; L-glutamine; L-arginine; L-serine; L-threonine; L-valine; L-tyrosine; and L-tryptophan. A enzyme protein retaining the activity of the wild type enzyme while having antioxidation properties against oxidation by hydrogen peroxide, and the like and a process for producing the same.

摘要翻译： 保留原始酶蛋白活性并具有抗氧化性的无硫原子的酶蛋白，其中酶蛋白的L-半胱氨酸和L-甲硫氨酸残基被18种L-氨基酸残基取代：L-丙氨酸; L-天冬氨酸; L-谷氨酸; L-苯丙氨酸 L-甘氨酸; L-组氨酸 L-异亮氨酸; L-赖氨酸 L-亮氨酸 L-天冬酰胺; L-脯氨酸 L-谷氨酰胺; L-精氨酸 L-丝氨酸 L-苏氨酸 L-缬氨酸 L-酪氨酸; 和L-色氨酸。 保留野生型酶的活性，同时具有抗过氧化氢氧化的抗氧化性的酶蛋白等及其制造方法。