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1. 发明授权

US07259022B2 Production of motif-specific and context-independent antibodies using peptide libraries as antigens 有权
标题翻译：使用肽文库作为抗原产生基序特异性和上下文相关抗体
公开(公告)号：US07259022B2
公开(公告)日：2007-08-21
申请号：US10014485
申请日：2001-11-13
申请人： Michael J. Comb , Yi Tan , Hui Zhang
发明人： Michael J. Comb , Yi Tan , Hui Zhang
IPC分类号： A61K38/00 , C07K16/00
CPC分类号： C07K16/00 , C07K16/44
摘要： A method is provided for producing motif-specific, context-independent antibodies which recognize a plurality of peptides or proteins within a genome that contain the same motif. The method includes the step of immunizing a host with a degenerate peptide library antigen featuring (i) a fixed target motif containing one or more invariant amino acids including at least one modified amino acid, and (ii) a plurality of degenerate amino adds flanking the motif. Motif-specific, context-independent antibodies produced by the disclosed method are also provided. The method encompasses motifs consisting of a single modified amino acid, as well as short motifs comprising multiple invariant amino acids including one or more modified amino acids, such as all or part of kinase consensus substrate motifs, protein-protein binding motifs, or other cell signaling motifs. Methods of using the antibodies, e.g. for genome-wide profiling, are also provided.
摘要翻译：提供了一种用于产生基序特异性上下文无关抗体的方法，其识别含有相同基序的基因组内的多个肽或蛋白质。该方法包括用简并肽文库抗原免疫宿主的步骤，其特征在于（i）含有一个或多个包含至少一个修饰的氨基酸的不变氨基酸的固定靶基序，和（ii）多个简并氨基酸，主题。还提供了通过公开的方法产生的基于特异性的，与环境无关的抗体。该方法包括由单个修饰的氨基酸组成的基序，以及包含多个不变氨基酸的短基序，包括一个或多个修饰的氨基酸，例如全部或部分激酶共有底物基序，蛋白质 - 蛋白质结合基序或其他细胞信号图案。使用抗体的方法，例如也提供了全基因组图谱。

2. 发明申请

US20110053242A1 Immunoaffinity isolation of modified peptides from complex mixtures 审中-公开
标题翻译：来自复杂混合物的修饰肽的免疫亲和分离
公开(公告)号：US20110053242A1
公开(公告)日：2011-03-03
申请号：US11823775
申请日：2007-06-28
申请人： John Rush , Hui Zhang , Xiangming Zha , Michael J. Comb , Yi Tan
发明人： John Rush , Hui Zhang , Xiangming Zha , Michael J. Comb , Yi Tan
IPC分类号： C12N11/00 , C12S3/14 , G01N33/566 , C07K1/14 , C07K16/18 , C07K16/00 , C07K16/40 , C07K1/22 , H01J49/26 , G01N30/72 , B01D11/02
CPC分类号： C07K16/18 , B01D15/3809 , B01D15/3828 , C07K1/22 , C07K16/44 , G01N30/7233 , G01N33/6842 , G01N33/6848 , G01N33/6854 , G01N2030/8831 , G01N2440/14
摘要： The invention provides methods for isolating a modified peptide from a complex mixture of peptides, the method comprising the steps of: (a) obtaining a proteinaceous preparation from an organism, wherein the preparation comprises modified peptides from two or more different proteins; (b) contacting the preparation with at least one immobilized modification-specific antibody; and (c) isolating at least one modified peptide specifically bound by the immobilized modification-specific antibody in step (b). The method may further comprise the step of (d) characterizing the modified peptide isolated in step (c) by mass spectrometry (MS), tandem mass spectrometry (MS-MS), and/or MS3 analysis, or the step of (e) utilizing a search program to substantially match the spectra obtained for the modified peptide during the characterization of step (d) with the spectra for a known peptide sequence, thereby identifying the parent protein(s) of the modified peptide. Also provided are an immunoaffinity isolation device comprising a modification-specific antibody, and antibodies against novel UFD1 and PTN6 phosphorylation sites.
摘要翻译：本发明提供了从复杂的肽混合物中分离修饰的肽的方法，所述方法包括以下步骤：（a）从生物体获得蛋白质制剂，其中所述制剂包含来自两种或更多种不同蛋白质的修饰肽; （b）使制剂与至少一种固定的修饰特异性抗体接触; 和（c）在步骤（b）中分离至少一种由固定的修饰特异性抗体特异性结合的修饰的肽。该方法还可以包括以下步骤：（d）通过质谱（MS），串联质谱（MS-MS）和/或MS3分析表征步骤（c）中分离的修饰肽，或（e）利用搜索程序将在步骤（d）表征期间获得的修饰肽的光谱与已知肽序列的光谱基本上匹配，从而鉴定修饰肽的亲本蛋白。还提供了包含修饰特异性抗体和针对新型UFD1和PTN6磷酸化位点的抗体的免疫亲和分离装置。

3. 发明申请

US20120244594A9 Immunoaffinity isolation of modified peptides from complex mixtures 审中-公开
标题翻译：来自复杂混合物的修饰肽的免疫亲和分离
公开(公告)号：US20120244594A9
公开(公告)日：2012-09-27
申请号：US11823775
申请日：2007-06-28
申请人： John Rush , Hui Zhang , Xiangming Zha , Michael J. Comb , Yi Tan
发明人： John Rush , Hui Zhang , Xiangming Zha , Michael J. Comb , Yi Tan
IPC分类号： C12N11/00 , G01N33/566 , C07K1/14 , C07K16/18 , B01D11/02 , C07K16/40 , C07K1/22 , H01J49/26 , G01N30/72 , C12S3/14 , C07K16/00
CPC分类号： C07K16/18 , B01D15/3809 , B01D15/3828 , C07K1/22 , C07K16/44 , G01N30/7233 , G01N33/6842 , G01N33/6848 , G01N33/6854 , G01N2030/8831 , G01N2440/14
摘要： The invention provides methods for isolating a modified peptide from a complex mixture of peptides, the method comprising the steps of: (a) obtaining a proteinaceous preparation from an organism, wherein the preparation comprises modified peptides from two or more different proteins; (b) contacting the preparation with at least one immobilized modification-specific antibody; and (c) isolating at least one modified peptide specifically bound by the immobilized modification-specific antibody in step (b). The method may further comprise the step of (d) characterizing the modified peptide isolated in step (c) by mass spectrometry (MS), tandem mass spectrometry (MS-MS), and/or MS3 analysis, or the step of (e) utilizing a search program to substantially match the spectra obtained for the modified peptide during the characterization of step (d) with the spectra for a known peptide sequence, thereby identifying the parent protein(s) of the modified peptide. Also provided are an immunoaffinity isolation device comprising a modification-specific antibody, and antibodies against novel UFD1 and PTN6 phosphorylation sites.
摘要翻译：本发明提供了从复杂的肽混合物中分离修饰的肽的方法，所述方法包括以下步骤：（a）从生物体获得蛋白质制剂，其中所述制剂包含来自两种或更多种不同蛋白质的修饰肽; （b）使制剂与至少一种固定的修饰特异性抗体接触; 和（c）在步骤（b）中分离至少一种由固定的修饰特异性抗体特异性结合的修饰的肽。该方法还可以包括以下步骤：（d）通过质谱（MS），串联质谱（MS-MS）和/或MS3分析表征步骤（c）中分离的修饰肽，或（e）利用搜索程序将在步骤（d）表征期间获得的修饰肽的光谱与已知肽序列的光谱基本上匹配，从而鉴定修饰肽的亲本蛋白。还提供了包含修饰特异性抗体和针对新型UFD1和PTN6磷酸化位点的抗体的免疫亲和分离装置。

4. 发明授权

US06982318B1 Production of motif-specific and context-independent antibodies using peptide libraries as antigens 有权
标题翻译：使用肽文库作为抗原产生基序特异性和上下文相关抗体
公开(公告)号：US06982318B1
公开(公告)日：2006-01-03
申请号：US09535364
申请日：2000-03-24
申请人： Michael J. Comb , Yi Tan
发明人： Michael J. Comb , Yi Tan
IPC分类号： C07K16/18
CPC分类号： C07K16/00 , C07K16/44
摘要： A class of motif-specific, context-independent antibodies that bind conserved signal transduction motifs, such as kinase consensus substrate motifs and protein-protein binding motifs, containing one or more modified amino acids, such as a phosphorylated amino acid, is provided. The antibodies bind a plurality of peptides or proteins that contain the modified motif. Context-independent antibodies specific for single modified residues, such as phosphothreonine, are also provided. Methods for producing and using such antibodies are provided.
摘要翻译：提供了一类与一个或多个修饰氨基酸如磷酸化氨基酸结合的保守信号转导基序（例如激酶共有底物基序和蛋白质 - 蛋白质结合基序）的基序特异性，上下文无关抗体。抗体结合多个含有修饰基序的肽或蛋白质。还提供了对单一修饰残基特异的上下文相关抗体，如磷酸苏氨酸。提供了生产和使用这些抗体的方法。

5. 发明授权

US06441140B1 Production of motif-specific and context-independent antibodies using peptide libraries as antigens 有权
标题翻译：使用肽文库作为抗原产生基序特异性和上下文相关抗体
公开(公告)号：US06441140B1
公开(公告)日：2002-08-27
申请号：US09148712
申请日：1998-09-04
申请人： Michael J. Comb , Yi Tan
发明人： Michael J. Comb , Yi Tan
IPC分类号： A61K3800
CPC分类号： C07K16/00 , C07K16/44
摘要： The present invention is related to a method for producing motif-specific, context-independent antibodies which are specific to at least one modified fixed amino acid residue in the context of variable surrounding amino acid or peptide sequences. The method is particularly useful in producing antibodies which recognize phosphorylated serine, threonine, and tyrosine, or acetylated lysine, as well as other modified amino acid-containing motifs of one or more amino acids.
摘要翻译：本发明涉及用于产生在可变周围氨基酸或肽序列的上下文中至少一个修饰的固定氨基酸残基特异性的基序特异性上下文无关抗体的方法。该方法特别可用于产生识别磷酸化丝氨酸，苏氨酸和酪氨酸或乙酰化赖氨酸的抗体，以及一种或多种氨基酸的其它经修饰的含氨基酸的基序。

6. 发明授权

US07344714B2 Production of motif-specific and context-independent antibodies using peptide libraries as antigens 有权
标题翻译：使用肽文库作为抗原产生基序特异性和上下文相关抗体
公开(公告)号：US07344714B2
公开(公告)日：2008-03-18
申请号：US10139841
申请日：2002-05-07
申请人： Michael J. Comb , Yi Tan
发明人： Michael J. Comb , Yi Tan
IPC分类号： A61K39/00 , A61K39/395 , A61K35/16 , C07K16/00
CPC分类号： C07K16/00 , C07K16/44
摘要： The present invention is related to a method for producing motif-specific, context-independent antibodies which are specific to at least one modified amino acid residue in the context of variable surrounding amino acid or peptide sequences. The method is particularly useful in producing antibodies which recognize phosphorylated serine, threonine, and tyrosine, or acetylated lysine, as well as other modified amino acids-containing motifs of one or more amino acids.
摘要翻译：本发明涉及用于产生在可变周围氨基酸或肽序列的上下文中至少一个修饰的氨基酸残基特异性的基序特异性上下文无关抗体的方法。该方法特别可用于产生识别磷酸化丝氨酸，苏氨酸和酪氨酸或乙酰化赖氨酸的抗体，以及一种或多种氨基酸的其它经修饰的含氨基酸的基序。

7. 发明授权

US07300753B2 Immunoaffinity isolation of modified peptides from complex mixtures 有权
标题翻译：来自复杂混合物的修饰肽的免疫亲和分离
公开(公告)号：US07300753B2
公开(公告)日：2007-11-27
申请号：US10777893
申请日：2004-02-12
申请人： John Rush , Hui Zhang , Xiangming Zha , Michael J. Comb , Yi Tan
发明人： John Rush , Hui Zhang , Xiangming Zha , Michael J. Comb , Yi Tan
IPC分类号： C12Q1/00 , C12Q1/37 , C12P21/06 , G01N33/543
CPC分类号： G01N33/6872 , C07K14/47 , C07K16/44 , C12Q1/25 , C12Q1/485 , G01N33/6842
摘要： The invention provides methods for isolating a modified peptide from a complex mixture of peptides, the method comprising the steps of: (a) obtaining a proteinaceous preparation from an organism, wherein the preparation comprises modified peptides from two or more different proteins; (b) contacting the preparation with at least one immobilized modification-specific antibody; and (c) isolating at least one modified peptide specifically bound by the immobilized modification-specific antibody in step (b). The method may further comprise the step of (d) characterizing the modified peptide isolated in step (c) by mass spectrometry (MS), tandem mass spectrometry (MS—MS), and/or MS3 analysis, or the step of (e) utilizing a search program to substantially match the spectra obtained for the modified peptide during the characterization of step (d) with the spectra for a known peptide sequence, thereby identifying the parent protein(s) of the modified peptide. Also provided are an immunoaffinity isolation device comprising a modification-specific antibody, and antibodies against novel UFD1 and PTN6 phosphorylation sites.
摘要翻译：本发明提供了从复杂的肽混合物中分离修饰的肽的方法，所述方法包括以下步骤：（a）从生物体获得蛋白质制剂，其中所述制剂包含来自两种或更多种不同蛋白质的修饰肽; （b）使制剂与至少一种固定的修饰特异性抗体接触; 和（c）在步骤（b）中分离至少一种由固定的修饰特异性抗体特异性结合的修饰的肽。该方法还可以包括以下步骤：（d）通过质谱法（MS），串联质谱（MS-MS）和/或MS 3分析表征步骤（c）中分离的修饰肽或（e）利用搜索程序将在步骤（d）表征期间获得的修饰肽的光谱与已知肽序列的光谱基本上匹配，从而鉴定修饰的肽的亲本蛋白质肽。还提供了包含修饰特异性抗体和针对新型UFD1和PTN6磷酸化位点的抗体的免疫亲和分离装置。

8. 发明授权

US07198896B2 Immunoaffinity isolation of modified peptides from complex mixtures 有权
公开(公告)号：US07198896B2
公开(公告)日：2007-04-03
申请号：US10175486
申请日：2002-06-19
申请人： John Rush , Hui Zhang , Xiangming Zha , Michael J. Comb , Yi Tan
发明人： John Rush , Hui Zhang , Xiangming Zha , Michael J. Comb , Yi Tan
IPC分类号： C12Q1/00 , C12Q1/37 , C12P21/06 , G01N33/543
CPC分类号： C07K16/44
摘要： The invention provides methods for isolating a modified peptide from a complex mixture of peptides, the method comprising the steps of: (a) obtaining a proteinaceous preparation from an organism, wherein the preparation comprises modified peptides from two or more different proteins; (b) contacting the preparation with at least one immobilized modification-specific antibody; and (c) isolating at least one modified peptide specifically bound by the immobilized modification-specific antibody in step (b). The method may further comprise the step of (d) characterizing the modified peptide isolated in step (c) by mass spectrometry (MS), tandem mass spectrometry (MS—MS), and/or MS3 analysis, or the step of (e) utilizing a search program to substantially match the spectra obtained for the modified peptide during the characterization of step (d) with the spectra for a known peptide sequence, thereby identifying the parent protein(s) of the modified peptide. Also provided are an immunoaffinity isolation device comprising a modification-specific antibody, and antibodies against novel UFD1 and PTN6 phosphorylation sites.

9. 发明授权

US07183385B2 Phospho-specific antibodies to Flt3 and uses thereof 有权
标题翻译： Flt3的磷酸特异性抗体及其用途
公开(公告)号：US07183385B2
公开(公告)日：2007-02-27
申请号：US10368912
申请日：2003-02-18
申请人： Michael J. Comb , Randall K. Wetzel , Jiong Wu , Katherine Crosby
发明人： Michael J. Comb , Randall K. Wetzel , Jiong Wu , Katherine Crosby
IPC分类号： C07K16/00 , C12P21/08 , G01N33/53
CPC分类号： C07K16/44 , C07K16/2896 , C07K16/40 , C07K2317/34 , G01N33/573 , G01N33/57426
摘要： The invention discloses two newly-discovered Flt3 phosphorylation sites, tyrosine 589 (Tyr589) and tyrosine 591 (Tyr591) in the intracellular domain, and provides antibodies, both polyclonal and monoclonal, that selectively bind to Flt3 when phosphorylated at these novel sites. Also provided are assays utilizing these reagents, including methods for determining the phosphorylation of Flt3 in a biological sample, selecting a patient suitable for Flt3 inhibitor therapy, profiling Flt3 activation in a test tissue, and identifying a compound that modulates phosphorylation of Flt3 in a test tissue, by using a detectable reagent, such as the disclosed antibodies, that binds to Flt3 when phosphorylated at Tyr589 or Tyr591. The sample or test tissue may be taken from a subject suspected of having cancer, such as acute myelogenous leukemia (AML).
摘要翻译：本发明公开了两个新发现的Flt3磷酸化位点，即细胞内结构域中的酪氨酸589（Tyr589）和酪氨酸591（Tyr591），并且提供了在这些新位点磷酸化时选择性结合Flt3的抗体，多克隆和单克隆抗体。还提供了使用这些试剂的测定法，包括用于测定生物样品中Flt3的磷酸化的方法，选择适合Flt3抑制剂治疗的患者，在测试组织中分析Flt3活化，以及鉴定在测试中调节Flt3的磷酸化的化合物通过使用在Tyr589或Tyr591磷酸化时与Flt3结合的可检测试剂，例如所公开的抗体。样品或测试组织可以从疑似患有癌症的受试者中获取，例如急性骨髓性白血病（AML）。

10. 发明授权

US09085609B2 Production of motif-specific and context-independent antibodies using peptide libraries as antigens 有权
标题翻译：使用肽文库作为抗原产生基序特异性和上下文相关抗体
公开(公告)号：US09085609B2
公开(公告)日：2015-07-21
申请号：US11484485
申请日：2006-07-11
申请人： Michael J. Comb , Peter Hornbeck , Patrick Li
发明人： Michael J. Comb , Yi Tan , Peter Hornbeck
IPC分类号： C07K16/00 , C07K16/18
CPC分类号： C07K16/18 , C07K16/00 , C07K16/44 , C07K2317/20
摘要： A method is provided for producing motif-specific, context-independent antibodies that recognize a plurality of peptides or proteins within a genome that contain the same post-translationally modified motif. The method includes the step of immunizing a host with a degenerate peptide library antigen featuring (i) a fixed target motif containing one or more invariant amino acids including at least one modified amino acid, and (ii) a plurality of degenerate amino acids flanking the motif. Motif-specific, context-independent antibodies produced by the disclosed method are also provided. The method encompasses motifs consisting of a single modified amino acid, as well as short motifs comprising multiple invariant amino acids including one or more modified amino acids, such as all or part of kinase consensus substrate motifs, protein-protein binding motifs, or other cell signaling motifs. Methods of using the antibodies, e.g. for genome-wide profiling, are also provided.
摘要翻译：提供了一种用于产生基序特异性，背景无关抗体的方法，其识别基因组内含有相同翻译后修饰基序的多个肽或蛋白质。该方法包括用简并肽文库抗原免疫宿主的步骤，其特征在于（i）含有一个或多个包含至少一个修饰的氨基酸的不变氨基酸的固定靶基序，和（ii）多个位于主题。还提供了通过公开的方法产生的基于特异性的，与环境无关的抗体。该方法包括由单个修饰的氨基酸组成的基序，以及包含多个不变氨基酸的短基序，包括一个或多个修饰的氨基酸，例如全部或部分激酶共有底物基序，蛋白质 - 蛋白质结合基序或其他细胞信号图案。使用抗体的方法，例如也提供了全基因组图谱。

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