公开(公告)号：US20090203034A1

公开(公告)日：2009-08-13

申请号：US12227318

申请日：2007-05-11

申请人： Ailan Guo ,  Roberto Polakiewicz ,  Kimberly Lee

发明人： Ailan Guo ,  Roberto Polakiewicz ,  Kimberly Lee

IPC分类号： G01N33/53 ,  C12Q1/48 ,  G01N33/536 ,  C07K16/18 ,  C07K14/47 ,  C07K7/06 ,  C07K7/08 ,  C12N5/16

CPC分类号： G01N33/6848 ,  C07K16/44 ,  G01N33/6842 ,  G01N33/6872 ,  G01N2800/2871

摘要： The invention discloses 99 novel phosphorylation sites identified in signal transduction proteins and pathways underlying human Brain Ischemia, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: protein kinases, adaptor/scaffold proteins, adhesion proteins, G proteins/GTPase/Guanine nucleotide exchange factors, Calcium binding proteins, cytoskeletal proteins, Channel proteins, Chaperone proteins, Helicases, Motor proteins, Translation proteins, RNA binding proteins, Ubiquitin conjugating system proteins, vesicle proteins and Receptor proteins.

摘要翻译： 本发明公开了在信号转导蛋白中识别的99个新的磷酸化位点和人类脑缺血的基因，并提供磷酸化位点特异性抗体和重同位素标记肽（AQUA肽），用于选择性检测和定量这些磷酸化位点/蛋白质，如 以及使用试剂用于此目的的方法。 鉴定的磷酸化位点之间是以下蛋白质类型中发生的位点：蛋白激酶，衔接/支架蛋白，粘附蛋白，G蛋白/ GTP酶/鸟嘌呤核苷酸交换因子，钙结合蛋白，细胞骨架蛋白，通道蛋白，伴侣蛋白，Helicases， 运动蛋白，翻译蛋白，RNA结合蛋白，泛素缀合系统蛋白，囊泡蛋白和受体蛋白。

公开(公告)号：US20100173322A1

公开(公告)日：2010-07-08

申请号：US12074876

申请日：2008-03-06

申请人： Roberto Polakiewicz ,  Ailan Guo ,  Albrecht Moritz ,  Kimberly Lee ,  Erik Spek ,  Charles Farnsworth ,  Francesco Boccalatte

发明人： Roberto Polakiewicz ,  Ailan Guo ,  Albrecht Moritz ,  Kimberly Lee ,  Erik Spek ,  Charles Farnsworth ,  Francesco Boccalatte

IPC分类号： G01N33/53 ,  C07K16/18 ,  G01N33/566

CPC分类号： C07K16/3061 ,  C07K2317/34 ,  G01N33/6842 ,  G01N33/6848

摘要： The invention discloses nearly 219 novel phosphorylation sites identified in signal transduction proteins and pathways underlying Anaplastic Large Cell Lymphoma (ALCL) involving the NPM-ALK translocation/fusion, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose.

摘要翻译： 本发明公开了在信号转导蛋白中鉴定的近219个新的磷酸化位点和涉及NPM-ALK易位/融合的间变性大细胞淋巴瘤（ALCL）的途径，并且提供磷酸化位点特异性抗体和重同位素标记肽（AQUA肽），用于 这些磷酸化位点/蛋白质的选择性检测和定量，以及用于这些目的的试剂的方法。

公开(公告)号：US20090298093A1

公开(公告)日：2009-12-03

申请号：US12226800

申请日：2007-04-27

申请人： Roberto Polakiewicz ,  Kimberly Lee ,  Ailan Guo ,  Matthew Stokes

发明人： Roberto Polakiewicz ,  Kimberly Lee ,  Ailan Guo ,  Matthew Stokes

IPC分类号： G01N33/573 ,  C12Q1/48 ,  C07K16/40

CPC分类号： C07K16/44 ,  C07B59/008

摘要： The invention discloses nearly 300 novel phosphorylation sites identified in signal transduction proteins and pathways underlying human ATM/ATR kinase signaling pathways, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection, profiling and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: DNA repair proteins, Adaptor/Scaffold proteins, Cell cycle regulation proteins, G Protein/GTPase Activating/Guanine Nucleotide Exchange Factor proteins, DNA binding proteins, DNA replication proteins, Kinases, Disease associated proteins proteins, Methyltransferase, Ubiquitin conjugating proteins, Proteases, Phosphatases, and Transcription proteins.

摘要翻译： 本发明公开了在信号转导蛋白中识别的近300个新的磷酸化位点和人类ATM / ATR激酶信号通路下的途径，并提供磷酸化位点特异性抗体和重同位素标记肽（AQUA肽）用于选择性检测，分析和定量 这些磷酸化位点/蛋白质，以及使用试剂用于此目的的方法。 鉴定的磷酸化位点是发生在以下蛋白质类型中的位点：DNA修复蛋白，适配器/支架蛋白，细胞周期调节蛋白，G蛋白/ GTP酶激活/鸟嘌呤核苷酸交换因子蛋白，DNA结合蛋白，DNA复制蛋白，激酶， 疾病相关蛋白质蛋白，甲基转移酶，泛素缀合蛋白，蛋白酶，磷酸酶和转录蛋白。

公开(公告)号：US07973134B2

公开(公告)日：2011-07-05

申请号：US11503096

申请日：2006-08-11

申请人： Albrecht Moritz ,  Kimberly Lee ,  John Rush ,  Roberto Polakiewicz

发明人： Albrecht Moritz ,  Kimberly Lee ,  John Rush ,  Roberto Polakiewicz

IPC分类号： C07K16/00 ,  C07K16/18

CPC分类号： G01N33/6842 ,  C07K16/44

摘要： The invention discloses 211 novel phosphorylation sites identified in signal transduction proteins and pathways underlying Anaplastic Large Cell Lymphoma (ALCL) involving the ALK-NPM translocation/fusion, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: Protein Kinases (including Receptor Tyrosine Kinases), Adaptor/Scaffold Proteins, Cellular Metabolism or Miscellaneous Enzymes, Oxidoreductases, Transcription Factors, Cytoskeletal Proteins, Translation Initiation Complexes, RNA Binding Proteins, Proteases, Acetyltransferases, G protein regulators/GTPases, Helicases, Apoptosis/Cell Cycle Regulation proteins, and Hydrolases.

摘要翻译： 本发明公开了在信号转导蛋白中鉴定的211个新的磷酸化位点和涉及ALK-NPM易位/融合的间变性大细胞淋巴瘤（ALCL）的途径，并且为磷酸化位点特异性抗体和重同位素标记的肽（AQUA肽）提供了 这些磷酸化位点/蛋白质的选择性检测和定量，以及使用试剂用于此目的的方法。 鉴定的磷酸化位点是以下蛋白质类型发生的位点：蛋白激酶（包括受体酪氨酸激酶），适配器/支架蛋白，细胞代谢或其他酶，氧化还原酶，转录因子，细胞骨架蛋白，翻译起始复合物，RNA结合蛋白， 蛋白酶，乙酰转移酶，G蛋白调节剂/ GTP酶，Helicases，细胞凋亡/细胞周期调节蛋白和水解酶。

公开(公告)号：US09920110B2

公开(公告)日：2018-03-20

申请号：US13416582

申请日：2012-03-09

申请人： Roberto Polakiewicz ,  Wan Cheung Cheung ,  John Edward Rush, II ,  Sean Andre Beausoleil

发明人： Roberto Polakiewicz ,  Wan Cheung Cheung ,  John Edward Rush, II ,  Sean Andre Beausoleil

IPC分类号： C07K1/14 ,  C07K16/00 ,  G01N33/50 ,  C07K16/08 ,  C07K16/18 ,  C07K16/44

CPC分类号： C07K16/00 ,  C07K16/082 ,  C07K16/18 ,  C07K16/44 ,  C07K2317/21 ,  C07K2317/56 ,  C07K2317/565

摘要： In some embodiments, the invention relates to methods for creating a monoclonal antibody that specifically binds to antigen. The method may start from a polyclonal population of antibodies such as a non-specific polyclonal population or a polyclonal population of antibodies that specifically bind to the antigen. The method includes obtaining nucleic acid molecules encoding heavy and light immunoglobulin chains (or variable regions thereof) of multiple immunoglobulins from an animal; obtaining mass spectra information of peptide fragments of a population of polyclonal immunoglobulins that specifically bind to an antigen of choice; comparing and/or correlating the mass spectra information of the peptide fragments of the polyclonal immunoglobulins with predicted mass spectra information of predicted amino acid sequences encoded by the nucleic acid molecules, and then assembling the heavy and light chains to create an antibody (or variable region thereof) that specifically binds to the antigen.

公开(公告)号：US20150031563A1

公开(公告)日：2015-01-29

申请号：US14235555

申请日：2012-07-27

申请人： Khanh Duc Huynh ,  Wan Cheung Cheung ,  Roberto Polakiewicz

发明人： Khanh Duc Huynh ,  Wan Cheung Cheung ,  Roberto Polakiewicz

IPC分类号： G01N33/53 ,  C12N9/52 ,  C07K16/18

CPC分类号： C12Q1/37 ,  C07K16/18 ,  C07K2319/43 ,  C07K2319/61 ,  C12N9/52 ,  C12Y304/2207 ,  G01N33/50 ,  G01N33/5306 ,  G01N33/542 ,  G01N33/56966 ,  G01N33/57492 ,  G01N2800/60

摘要： The disclosure provides methods for detecting the concurrent presence of at least two targets within a biological sample. The method includes contacting said biological sample with a first binding agent, said first binding agent operably linked to a first sortase molecule, wherein said first binding agent specifically binds to a first target; contacting said biological sample with a second binding agent, said second binding agent operably linked to a first sortase recognition sequence peptide, wherein said second binding agent specifically binds to a second target; adding a sortase substrate under conditions where a first sortase-mediated ligation of the sortase substrate to the first sortase recognition sequence will produce a ligation product, and detecting the ligation product, wherein detection of said ligation product indicates the concurrent presence of the first target and the second target in the biological sample. Also disclosed are kits comprising reagents for performing the methods as claimed.

摘要翻译： 本公开提供了用于检测生物样品中至少两个靶的同时存在的方法。 该方法包括使所述生物样品与第一结合剂接触，所述第一结合剂可操作地连接到第一分选酶分子，其中所述第一结合剂特异性结合至第一靶标; 使所述生物样品与第二结合剂接触，所述第二结合剂可操作地连接到第一分选酶识别序列肽，其中所述第二结合剂特异性结合第二靶标; 在分选酶底物与第一分选酶识别序列的第一分选酶介导的连接将产生连接产物并检测连接产物的条件下添加分选酶底物，其中所述连接产物的检测表明第一靶标的同时存在， 生物样本中的第二个目标。 还公开了包含用于执行所要求保护的方法的试剂的试剂盒。

公开(公告)号：US07935790B2

公开(公告)日：2011-05-03

申请号：US11503336

申请日：2006-08-11

申请人： Albrecht Moritz ,  Roberto Polakiewicz ,  John Edward Rush ,  Kimberly Lee

发明人： Albrecht Moritz ,  Roberto Polakiewicz ,  John Edward Rush ,  Kimberly Lee

IPC分类号： C07K16/00 ,  C07K16/18

CPC分类号： G01N33/6872 ,  C12Q1/25 ,  C12Q1/485 ,  G01N33/6842

摘要： The invention discloses 95 novel phosphorylation sites identified in signal transduction proteins and pathways downstream of the T-cell receptor, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: Actin Binding proteins, Adaptor/Scaffold proteins, Adhesion proteins, Calcium-binding proteins, Cell Cycle Regulation or Channel proteins, Chaperones, Cofactor proteins, Cytoskeletal proteins, DNA Binding proteins, G protein or GTPase Activating proteins, Ligases, Lipid Kinases and Binding proteins, Oxidoreductases, Protein Kinases, Protein Phosphatases, Receptor proteins, RNA Binding proteins, Transcription Factor/Initiation Complex proteins, Transcription Coactivator/Corepressor proteins, Translation Initiation Complex proteins, Ubitquitin Conjugating System proteins, and Vesicle proteins.

摘要翻译： 本发明公开了在T细胞受体下游的信号转导蛋白和途径中鉴定的95个新的磷酸化位点，并提供磷酸化位点特异性抗体和重同位素标记肽（AQUA肽），用于选择性检测和定量这些磷酸化位点/ 蛋白质，以及使用试剂用于此目的的方法。 鉴定的磷酸化位点是发生在以下蛋白质类型中的位点：肌动蛋白结合蛋白，衔接/支架蛋白，粘附蛋白，钙结合蛋白，细胞周期调节或通道蛋白，伴侣蛋白，辅因子蛋白，细胞骨架蛋白，DNA结合蛋白， G蛋白或GTPase激活蛋白，连接酶，脂质激酶和结合蛋白，氧化还原酶，蛋白激酶，蛋白磷酸酶，受体蛋白，RNA结合蛋白，转录因子/起始复合蛋白​​，转录共激活因子/ Corepressor蛋白，翻译起始复合蛋白​​，Ubitquitin缀合 系统蛋白和囊泡蛋白。

公开(公告)号：US07731964B2

公开(公告)日：2010-06-08

申请号：US10694874

申请日：2003-10-28

申请人： Roberto Polakiewicz ,  Jiong Wu ,  Yu Li

发明人： Roberto Polakiewicz ,  Jiong Wu ,  Yu Li

IPC分类号： A61K39/00 ,  A61K39/395 ,  C07K16/00

CPC分类号： C07K16/44 ,  C07K16/18 ,  G01N33/74

摘要： The invention discloses newly-discovered phosphorylation sites in human IRS-1 and IRS-2, serine 1101 (Ser1101) and serine 1149 (Ser1149) respectively, and provides antibodies, both polyclonal and monoclonal, that selectively bind to IRS-1 and/or IRS-2 when phosphorylated at these respective sites, but do not bind to IRS-1 and/or IRS-2 when not phosphorylated at these respective sites. The sites are relevant to insulin-resistance in type 2 diabetes. Also provided are methods for determining the phosphorylation of IRS-1/2 or activity of PKC theta in a biological sample, by using a detectable reagent, such as the disclosed antibodies, that binds to IRS-1/2 only when phosphorylated at Ser1101/Ser1149. Kits comprising the phosphor-IRS-1/2 (Ser1101/1149) antibodies of the invention are also provided.

摘要翻译： 本发明公开了分别在人IRS-1和IRS-2，丝氨酸1101（Ser1101）和丝氨酸1149（Ser1149）中新发现的磷酸化位点，并提供选择性结合IRS-1和/或 IRS-2在这些相应位点处磷酸化，但当不在这些相应位点被磷酸化时不结合IRS-1和/或IRS-2。 这些部位与2型糖尿病中的胰岛素抵抗有关。 还提供了通过使用可检测的试剂，例如所公开的抗体，其仅在Ser1101 / 1细胞磷酸化时与IRS-1/2结合才能确定生物样品中IRS-1/2的磷酸化或PKCθ的活性的方法， Ser1149。 还提供了包含本发明的磷光体IRS-1/2（Ser1101 / 1149）抗体的试剂盒。

公开(公告)号：US20080248490A1

公开(公告)日：2008-10-09

申请号：US12074224

申请日：2008-02-29

申请人： Roberto Polakiewicz ,  Valerie Goss ,  Albrecht Moritz ,  Ting-Lei Gu ,  Kimberly Lee

发明人： Roberto Polakiewicz ,  Valerie Goss ,  Albrecht Moritz ,  Ting-Lei Gu ,  Kimberly Lee

IPC分类号： C07K16/18 ,  G01N33/53

CPC分类号： G01N33/6842 ,  C07K16/3061 ,  C07K16/44 ,  C07K2317/34 ,  G01N33/57426 ,  G01N2458/15

摘要： The invention discloses nearly 288 novel phosphorylation sites identified in signal transduction proteins and pathways underlying human Leukemia, and provides phosphorylation-site specific antibodies and heavy-isotope labeled peptides (AQUA peptides) for the selective detection and quantification of these phosphorylated sites/proteins, as well as methods of using the reagents for such purpose. Among the phosphorylation sites identified are sites occurring in the following protein types: Adaptor/Scaffold proteins, Cytoskeletal proteins, Cellular Metabolism enzymes, G Protein/GTPase Activating/Guanine Nucleotide Exchange Factor proteins, Immunoglobulin Superfamily proteins, Inhibitor proteins, Lipid Kinases, Nuclear DNA Repair/RNA Binding/Transcription proteins, Serine/Threonine Protein Kinases, Tyrosine Kinases, Protein Phosphatases, and Translation/Transporter proteins.

摘要翻译： 本发明公开了在信号转导蛋白中鉴定的近288个新的磷酸化位点和人类白血病潜在的途径，并提供磷酸化位点特异性抗体和重同位素标记肽（AQUA肽），用于选择性检测和定量这些磷酸化位点/蛋白，如 以及使用试剂用于此目的的方法。 所鉴定的磷酸化位点是以下蛋白质类型的位点：适配器/支架蛋白，细胞骨架蛋白，细胞代谢酶，G蛋白/ GTP酶激活/鸟嘌呤核苷酸交换因子蛋白，免疫球蛋白超家族蛋白，抑制蛋白，脂质激酶，核DNA 修复/ RNA结合/转录蛋白，丝氨酸/苏氨酸蛋白激酶，酪氨酸激酶，蛋白磷酸酶和翻译/转运蛋白。

公开(公告)号：US09453845B2

公开(公告)日：2016-09-27

申请号：US12931445

申请日：2011-02-01

申请人： Albrecht Moritz ,  John Edward Rush, II ,  Roberto Polakiewicz

发明人： Albrecht Moritz ,  John Edward Rush, II ,  Roberto Polakiewicz

IPC分类号： C12Q1/70 ,  B01D59/44 ,  H01J49/00 ,  G01N15/06 ,  G01N33/00 ,  G01N33/48 ,  G01N33/53 ,  G01N33/68

CPC分类号： G01N33/6848 ,  B01D15/12 ,  B01D15/325 ,  B01D15/3809 ,  B01D15/3823 ,  B01D15/422 ,  G01N30/7233 ,  G01N2560/00 ,  G01N2570/00 ,  H01J49/00

摘要： The invention relates to a method for determining the presence of at least one distinct polypeptide in a biological sample comprising contacting the biological sample with a hydrolyzing agent, wherein the hydrolyzing agent is capable of hydrolyzing the distinct polypeptide in a sequence-specific manner such that at least one distinct peptide having a predetermined peptide measured accurate mass would result if the at least one distinct polypeptide were present in the biological sample, to obtain a hydrolyzed sample; bringing the hydrolyzed sample in contact with a substrate comprising at least one immobilized binding partner, wherein the at least one immobilized binding partner is capable of specifically binding the distinct peptide; removing the hydrolyzed sample from the substrate in a manner such that the distinct peptide would remain bound to the immobilized binding partner; contacting the substrate with an elution solution, wherein the distinct peptide would dissociate from the immobilized binding partner into the elution solution; subjecting a portion of the elution solution to liquid chromatography to segregate a plurality of molecules in the portion of the elution solution to obtain sorted molecules; determining the measured accurate mass of at least one sorted molecule present in the elution solution; and determining the presence of the at least one distinct polypeptide in the biological sample when a measured accurate mass of at least one molecule is substantially equal to the predetermined peptide measured accurate mass.

摘要翻译： 本发明涉及一种用于确定生物样品中至少一种不同多肽的存在的方法，包括使生物样品与水解试剂接触，其中水解试剂能够以序列特异性方式水解不同的多肽， 如果至少一种不同的多肽存在于生物样品中，则得到具有测定精确质量的预定肽的至少一种不同的肽，以获得水解样品; 使水解的样品与包含至少一个固定的结合配偶体的底物接触，其中至少一个固定的结合配偶体能够特异性结合不同的肽; 以使得不同的肽保持与固定的结合配偶体结合的方式从底物中除去水解的样品; 使所述底物与洗脱溶液接触，其中所述不同的肽将从固定的结合配偶体解离成洗脱溶液; 使洗脱溶液的一部分进行液相色谱分离洗脱溶液部分中的多个分子，得到分选的分子; 确定洗脱溶液中存在的至少一种分选分子的测量精确质量; 以及当所测量的至少一个分子的精确质量基本上等于所测量的精确质量的预定肽时，确定生物样品中至少一种不同多肽的存在。