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1. 发明申请

US20090023898A1 METHODS AND COMPOSITIONS FOR PROTEIN PURIFICATION 审中-公开
标题翻译：蛋白质纯化的方法和组合物
公开(公告)号：US20090023898A1
公开(公告)日：2009-01-22
申请号：US12203894
申请日：2008-09-03
申请人： Grigoriy S. Tchaga , George G. Jokhadze
发明人： Grigoriy S. Tchaga , George G. Jokhadze
IPC分类号： C07K1/14
CPC分类号： C07K7/06 , C07K7/08 , C07K14/001 , C07K2319/00 , C07K2319/02 , C07K2319/21 , C07K2319/60 , C12N15/62
摘要： The present invention provides methods of purifying proteins that include a metal ion affinity peptide. The methods generally involve contacting a fusion protein that includes a metal ion affinity peptide with at least two different metal ion chelating resins. In certain representative embodiments, the methods include contacting a fusion protein with a first metal ion chelate resin having a first immobilized metal ion; eluting any bound protein from the first metal ion chelate resin, to produce an eluate; contacting the eluate with a second metal ion chelate resin having a second immobilized metal ion; and eluting any bound protein from the second metal ion chelate resin. Also provided are kits for use in practicing the subject methods. The subject methods find use in a variety of protein purification applications.
摘要翻译：本发明提供了纯化包括金属离子亲和肽的蛋白质的方法。所述方法通常包括使包含金属离子亲和肽的融合蛋白与至少两种不同的金属离子螯合树脂接触。在某些代表性实施方案中，所述方法包括使融合蛋白与具有第一固定金属离子的第一金属离子螯合树脂接触; 从第一金属离子螯合树脂洗脱任何结合的蛋白质，以产生洗脱液; 使洗脱液与具有第二固定化金属离子的第二金属离子螯合树脂接触; 并从第二金属离子螯合树脂洗脱任何结合的蛋白质。还提供了用于实践主题方法的套件。主题方法可用于各种蛋白质纯化应用。

2. 发明授权

US07723125B2 Highly sensitive proteomic analysis methods, and kits and systems for practicing the same 有权
标题翻译：高度敏感的蛋白质组学分析方法，以及实践相同的试剂盒和系统
公开(公告)号：US07723125B2
公开(公告)日：2010-05-25
申请号：US11850474
申请日：2007-09-05
申请人： Grigoriy S. Tchaga
发明人： Grigoriy S. Tchaga
IPC分类号： G01N33/543
CPC分类号： G01N33/552 , B01J2219/00585 , B01J2219/00596 , B01J2219/00605 , B01J2219/00612 , B01J2219/00626 , B01J2219/0063 , B01J2219/00637 , B01J2219/00659 , B01J2219/0072 , C03C17/30 , C03C17/3405 , G01N33/54353 , G01N33/582 , G01N33/6803 , G01N2400/46
摘要： Methods of determining whether a sample includes one or more analytes, particularly proteinaceous analytes, of interest are provided. In the subject methods, an array of binding agents, where each binding agent includes an epitope binding domain of an antibody, is contacted with the sample. In many embodiments, contact occurs in the presence of a metal ion chelating polysaccharide, e.g., a pectin. Following contact, the presence of binding complexes on the array surface are detected and the resultant data is employed to determine whether the sample includes the one or more analytes of interest. Also provided are kits, systems and other compositions of matter for practicing the subject methods. The subject methods and compositions find use in a variety of applications, including proteomic applications such as protein expression analysis, e.g., differential protein expression profiling.
摘要翻译：提供了确定样品是否包含一种或多种感兴趣的分析物，特别是蛋白质分析物的方法。在本发明方法中，将每种结合剂包括抗体的表位结合结构域的结合剂阵列与样品接触。在许多实施方案中，接触发生在金属离子螯合多糖例如果胶的存在下。接触后，检测阵列表面上结合复合物的存在，并使用得到的数据来确定样品是否包含一种或多种感兴趣的分析物。还提供了用于实践主题方法的试剂盒，系统和其它物质组合物。本发明的方法和组合物可用于各种应用，包括蛋白质组学应用，例如蛋白质表达分析，例如差异蛋白质表达谱。

3. 发明授权

US07176298B2 Polynucleotides encoding metal ion affinity peptides and related products 有权
标题翻译：编码金属离子亲和肽和相关产品的多核苷酸
公开(公告)号：US07176298B2
公开(公告)日：2007-02-13
申请号：US09858332
申请日：2001-05-15
申请人： Grigoriy S. Tchaga , George G. Jokhadze
发明人： Grigoriy S. Tchaga , George G. Jokhadze
IPC分类号： C07H21/02 , C07H21/04 , C12N15/63 , C12N1/20 , C12N15/00
CPC分类号： C07K7/06 , C07K7/08 , C07K14/001 , C07K2319/00 , C07K2319/02 , C07K2319/21 , C07K2319/60 , C12N15/62
摘要： The present invention provides metal ion affinity peptides, fusion proteins comprising metal ion affinity peptides, and polynucleotides encoding the fusion proteins. A feature of the subject invention is that the metal ion affinity peptide has a formula selected from the group consisting of: formula 1: (His-X1-X2)n1-(His-X3-X4-X5)n2-(His-X6)n3, wherein each of X1 and X2 is independently an amino acid with an aliphatic or an amide side chain, each of X3, X4, X5 is independently an amino acid with a basic side chain (except His) or an acidic side chain, each X6 is an amino acid with an aliphatic or an amide side chain, n1 and n2 are each independently 1–3, and n3 is 1–5; formula 2: (His-Asn)n, where n=3 to 10; and formula 3: (His-X1-X2)n, wherein each of X1 and X2 is an amino acid having an acidic side chain, and n=3 to 10. The invention further provides recombinant vectors comprising subject polynucleotides, and host cells comprising the recombinant vectors. The invention further provides methods and kits for purifying a fusion protein comprising a metal ion affinity peptide.
摘要翻译：本发明提供了金属离子亲和肽，包含金属离子亲和肽的融合蛋白和编码融合蛋白的多核苷酸。本发明的一个特征是金属离子亲和肽具有选自以下的式：式1：（His-X 1 -X 2 -X 2）（His-X 3 -X 4 -X 5）N 2 - （His-X 3 -X 4 - （His-X 6）n 3，其中X 1和X 2各自独立地是具有 X 3，X 4，X 5各自独立地是具有碱性侧链的氨基酸（脂肪族或酰胺侧链）除了His）或酸性侧链，每个X 6是具有脂族或酰胺侧链的氨基酸，n1和n2各自独立地为1-3，n3为1-5; 式2：（His-Asn）n N，其中n = 3至10; 和式3：（His-X 1 -X 2）n N，其中X 1和X 2中的每一个具有酸性侧链的氨基酸，n = 3〜10。本发明还提供包含受试多核苷酸的重组载体和包含重组载体的宿主细胞。本发明还提供了用于纯化包含金属离子亲和肽的融合蛋白的方法和试剂盒。

4. 发明申请

US20090099032A1 HIGHLY SENSITIVE PROTEOMIC ANALYSIS METHODS, AND KITS AND SYSTEMS FOR PRACTICING THE SAME 有权
标题翻译：高敏感性分析方法及其实施方法及其系统
公开(公告)号：US20090099032A1
公开(公告)日：2009-04-16
申请号：US11850474
申请日：2007-09-05
申请人： Grigoriy S. Tchaga
发明人： Grigoriy S. Tchaga
IPC分类号： C40B30/04
CPC分类号： G01N33/552 , B01J2219/00585 , B01J2219/00596 , B01J2219/00605 , B01J2219/00612 , B01J2219/00626 , B01J2219/0063 , B01J2219/00637 , B01J2219/00659 , B01J2219/0072 , C03C17/30 , C03C17/3405 , G01N33/54353 , G01N33/582 , G01N33/6803 , G01N2400/46
摘要： Methods of determining whether a sample includes one or more analytes, particularly proteinaceous analytes, of interest are provided. In the subject methods, an array of binding agents, where each binding agent includes an epitope binding domain of an antibody, is contacted with the sample. In many embodiments, contact occurs in the presence of a metal ion chelating polysaccharide, e.g., a pectin. Following contact, the presence of binding complexes on the array surface are detected and the resultant data is employed to determine whether the sample includes the one or more analytes of interest. Also provided are kits, systems and other compositions of matter for practicing the subject methods. The subject methods and compositions find use in a variety of applications, including proteomic applications such as protein expression analysis, e.g., differential protein expression profiling.
摘要翻译：提供了确定样品是否包含一种或多种感兴趣的分析物，特别是蛋白质分析物的方法。在本发明方法中，将每种结合剂包括抗体的表位结合结构域的结合剂阵列与样品接触。在许多实施方案中，接触发生在金属离子螯合多糖例如果胶的存在下。接触后，检测阵列表面上结合复合物的存在，并使用得到的数据来确定样品是否包含一种或多种感兴趣的分析物。还提供了用于实践主题方法的试剂盒，系统和其它物质组合物。本发明的方法和组合物可用于各种应用，包括蛋白质组学应用，例如蛋白质表达分析，例如差异蛋白质表达谱。

5. 发明授权

US07354721B2 Highly sensitive proteomic analysis methods, and kits and systems for practicing the same 失效
标题翻译：高度敏感的蛋白质组学分析方法，以及实践相同的试剂盒和系统
公开(公告)号：US07354721B2
公开(公告)日：2008-04-08
申请号：US09960716
申请日：2001-09-21
申请人： Grigoriy S. Tchaga
发明人： Grigoriy S. Tchaga
IPC分类号： G01N33/53
CPC分类号： G01N33/552 , B01J2219/00585 , B01J2219/00596 , B01J2219/00605 , B01J2219/00612 , B01J2219/00626 , B01J2219/0063 , B01J2219/00637 , B01J2219/00659 , B01J2219/0072 , C03C17/30 , C03C17/3405 , G01N33/54353 , G01N33/582 , G01N33/6803 , G01N2400/46
摘要： Methods of determining whether a sample includes one or more analytes, particularly proteinaceous analytes, of interest are provided. In the subject methods, an array of binding agents, where each binding agent includes an epitope binding domain of an antibody, is contacted with the sample. In many embodiments, contact occurs in the presence of a metal ion chelating polysaccharide, e.g., a pectin. Following contact, the presence of binding complexes on the array surface are detected and the resultant data is employed to determine whether the sample includes the one or more analytes of interest. Also provided are kits, systems and other compositions of matter for practicing the subject methods. The subject methods and compositions find use in a variety of applications, including proteomic applications such as protein expression analysis, e.g., differential protein expression profiling.
摘要翻译：提供了确定样品是否包含一种或多种感兴趣的分析物，特别是蛋白质分析物的方法。在本发明方法中，将每种结合剂包括抗体的表位结合结构域的结合剂阵列与样品接触。在许多实施方案中，接触发生在金属离子螯合多糖例如果胶的存在下。接触后，检测阵列表面上结合复合物的存在，并使用得到的数据来确定样品是否包含一种或多种感兴趣的分析物。还提供了用于实践主题方法的试剂盒，系统和其它物质组合物。本发明的方法和组合物可用于各种应用，包括蛋白质组学应用，例如蛋白质表达分析，例如差异蛋白质表达谱。

6. 发明授权

US06703498B2 Water-soluble polymeric metal ion affinity compositions and methods for using the same 有权
标题翻译：水溶性聚合物金属离子亲和性组合物及其使用方法
公开(公告)号：US06703498B2
公开(公告)日：2004-03-09
申请号：US10176955
申请日：2002-06-20
申请人： Grigoriy S. Tchaga
发明人： Grigoriy S. Tchaga
IPC分类号： C08B3702
CPC分类号： C08B37/0021 , C07C229/76 , C07K1/22
摘要： Water-soluble metal ion affinity compounds and methods for using the same are provided. The subject compounds include an aspartate based metal chelating ligand bonded to a water-soluble polymeric substrate, where the ligand is complexed with a metal ion. In certain embodiments, the subject compounds further include a member of a signal producing system, e.g., a directly or an indirectly detectable label moiety. Also provided are water-insoluble supports having the subject compounds present on, e.g., immobilized on, at least one surface thereof. The subject compounds find use in a variety of different applications, including analyte detection and analyte purification applications.
摘要翻译：提供了水溶性金属离子亲和性化合物及其使用方法。主题化合物包括与水溶性聚合物底物结合的天冬氨酸盐基金属螯合配体，其中配体与金属离子络合。在某些实施方案中，本发明化合物还包括信号产生系统的成员，例如直接或间接可检测的标记部分。还提供了具有例如固定在其至少一个表面上的主题化合物的水不溶性载体。主题化合物可用于各种不同的应用，包括分析物检测和分析物纯化应用。

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