公开(公告)号：US20220137066A1

公开(公告)日：2022-05-05

申请号：US17399434

申请日：2021-08-11

申请人： AmberGen, Inc.

发明人： Mark J. Lim ,  Gargey Yagnik ,  Kenneth J. Rothschild

IPC分类号： G01N33/68

摘要： The field of this invention relates to immunohistochemistry (IHC) and in situ hybridization (ISH) for the targeted detection and mapping of biomolecules (e.g., proteins and miRNAs) in tissues or cells for example, for research use and for clinical use such by pathologists (e.g., biomarker analyses of a resected tumor or tumor biopsy). In particular, the use of mass spectrometric imaging (MSI) as a mode to detect and map the biomolecules in tissues or cells for example. More specifically, the field of this invention relates to photocleavable mass-tag reagents which are attached to probes such as antibodies and nucleic acids and used to achieve multiplex immunohistochemistry and in situ hybridization, with MSI as the mode of detection/readout. Probe types other than antibodies and nucleic acids are also covered in the field of invention, including but not limited to carbohydrate-binding proteins (e.g., lectins), receptors and ligands. Finally, the field of the invention also encompasses multi-omic MSI procedures, where MSI of photocleavable mass-tag probes is combined with other modes of MSI, such as direct label-free MSI of endogenous biomolecules from the biospecimen (e.g., tissue), whereby said biomolecules can be intact or digested (e.g., chemically digested or by enzyme).

公开(公告)号：US20220135518A1

公开(公告)日：2022-05-05

申请号：US17399461

申请日：2021-08-11

申请人： AmberGen, Inc.

发明人： Mark J. Lim ,  Gargey Yagnik ,  Kenneth J. Rothschild

IPC分类号： C07C271/20 ,  G01N33/68

摘要： The field of this invention relates to immunohistochemistry (IHC) and in situ hybridization (ISH) for the targeted detection and mapping of biomolecules (e.g., proteins and miRNAs) in tissues or cells for example, for research use and for clinical use such by pathologists (e.g., biomarker analyses of a resected tumor or tumor biopsy). In particular, the use of mass spectrometric imaging (MSI) as a mode to detect and map the biomolecules in tissues or cells for example. More specifically, the field of this invention relates to photocleavable mass-tag reagents which are attached to probes such as antibodies and nucleic acids and used to achieve multiplex immunohistochemistry and in situ hybridization, with MSI as the mode of detection/readout. Probe types other than antibodies and nucleic acids are also covered in the field of invention, including but not limited to carbohydrate-binding proteins (e.g., lectins), receptors and ligands. Finally, the field of the invention also encompasses multi-omic MSI procedures, where MSI of photocleavable mass-tag probes is combined with other modes of MSI, such as direct label-free MSI of endogenous biomolecules from the biospecimen (e.g., tissue), whereby said biomolecules can be intact or digested (e.g., chemically digested or by enzyme).

公开(公告)号：US08906700B2

公开(公告)日：2014-12-09

申请号：US13553472

申请日：2012-07-19

申请人： Mark J. Lim ,  Kenneth J. Rothschild

发明人： Mark J. Lim ,  Kenneth J. Rothschild

IPC分类号： G01N33/543 ,  G01N33/53 ,  C40B30/04

CPC分类号： G01N33/54306 ,  C12Q1/6806 ,  G01N33/54353 ,  G01N33/54393

摘要： Methods are described for phototransferring a compound from a first surface to a second surface. Compounds are described with photocleavable linkers. Compounds attached to a first surface through a photocleavable linker are put in proximity (or contact) with a second surface, and then phototransferred to the second surface upon exposure to electromagnetic radiation. Illuminating the compound with radiation photocleaves the compound from the first surface and transfers the compound to the second surface.

摘要翻译： 描述了将化合物从第一表面转移到第二表面的方法。 用可光洁化的接头描述化合物。 通过光可透洗接头连接到第一表面的化合物与第二表面接近（或接触），然后在暴露于电磁辐射时转移到第二表面。 用辐射照射化合物使化合物从第一表面光化，并将化合物转移到第二表面。

公开(公告)号：US20100311042A1

公开(公告)日：2010-12-09

申请号：US12291178

申请日：2008-11-06

申请人： Mark J. Lim ,  Kenneth J. Rothschild

发明人： Mark J. Lim ,  Kenneth J. Rothschild

IPC分类号： C12Q1/68 ,  C12P19/34

CPC分类号： C12P19/34 ,  C12Q1/6844

摘要： Methods of amplifying nucleic acid are described. Primers on a solid support, e.g. a population of beads, are employed. A population of nucleic acid template molecules, wherein said nucleic acid template molecules have been treated with bisulfite, is amplified so as to create loaded beads comprising immobilized amplified nucleic acid.

摘要翻译： 描述了扩增核酸的方法。 固体支持物上的引物，例如 一群珠子被雇用。 扩增核酸模板分子群体，其中所述核酸模板分子已经用亚硫酸氢盐处理，以产生包含固定化扩增核酸的负载珠。

公开(公告)号：US07807407B2

公开(公告)日：2010-10-05

申请号：US10903612

申请日：2004-07-30

申请人： Alex M. Garvin ,  Sadanand Gite ,  Vladislav B. Bergo ,  Kenneth J. Rothschild

发明人： Alex M. Garvin ,  Sadanand Gite ,  Vladislav B. Bergo ,  Kenneth J. Rothschild

IPC分类号： C12P21/06 ,  C12P19/34 ,  C12N9/50

CPC分类号： G01N33/6848

摘要： This invention relates to the detection and analysis by mass spec of nascent proteins, and in particular truncated proteins, translated within cellular or cell-free translation systems. N-terminal and C-terminal epitopes introduced into these nascent proteins permit rapid and efficient isolation, as well as a mass difference.

摘要翻译： 本发明涉及通过新生蛋白的质谱检测和分析，特别是在细胞或无细胞翻译系统内翻译的截短蛋白质。 引入这些新生蛋白质的N-末端和C-末端表位允许快速和有效的分离以及质量差异。

公开(公告)号：US20100203651A1

公开(公告)日：2010-08-12

申请号：US12290447

申请日：2008-10-30

申请人： Mark J. Lim ,  Kenneth J. Rothschild

发明人： Mark J. Lim ,  Kenneth J. Rothschild

IPC分类号： G01N33/543 ,  C07K1/107 ,  C07K16/00 ,  C07H1/00 ,  C08G63/48

CPC分类号： C12P21/02 ,  B01J2219/00459 ,  B01J2219/005 ,  B01J2219/00576 ,  B01J2219/00596 ,  B01J2219/00648 ,  B01J2219/00722 ,  B01J2219/00725 ,  C12N15/1075 ,  C40B40/08 ,  C40B50/06 ,  C40B80/00

摘要： Methods are described for phototransferring a compound from a first surface to a second surface. Compounds are described with photocleavable linkers. Compounds attached to a first surface through a photocleavable linker are put in proximity (or contact) with a second surface, and then phototransferred to the second surface upon exposure to electromagnetic radiation. Illuminating the compound with radiation photocleaves the compound from the first surface and transfers the compound to the second surface.

摘要翻译： 描述了将化合物从第一表面转移到第二表面的方法。 用可光洁化的接头描述化合物。 通过光可透洗接头连接到第一表面的化合物与第二表面接近（或接触），然后在暴露于电磁辐射时转移到第二表面。 用辐射照射化合物使化合物从第一表面光化，并将化合物转移到第二表面。

公开(公告)号：US20090263868A1

公开(公告)日：2009-10-22

申请号：US12485213

申请日：2009-06-16

申请人： Jerzy Olejnik ,  Edyta Krzymanska-Olejnik ,  Kenneth J. Rothschild

发明人： Jerzy Olejnik ,  Edyta Krzymanska-Olejnik ,  Kenneth J. Rothschild

IPC分类号： C12P19/34 ,  C07H21/02 ,  C07H21/04

CPC分类号： A61K49/0032 ,  A61K49/0021 ,  A61K49/0052 ,  C07H19/00 ,  C07H19/04 ,  C07H21/02 ,  C07H21/04

摘要： Labelled nucleotides and polynucleotides useful in the sequencing of nucleic acids are described. Methods of preparing photocleavable marker nucleotides and photocleavable marker-polynucleotide conjugates are described. Such photocleavable marker nucleotides can be incorporated into nucleic acid so as to create photocleavable marker-polynucleotide conjugates.

摘要翻译： 描述了用于核酸测序的标记核苷酸和多核苷酸。 描述了制备可光切割标记核苷酸和可光切割标记 - 多核苷酸缀合物的方法。 可将这种可光切割的标记核苷酸掺入核酸中，以产生可光切割的标记 - 多核苷酸缀合物。

公开(公告)号：US07524941B2

公开(公告)日：2009-04-28

申请号：US11900323

申请日：2007-09-11

申请人： Jerzy Olejnik ,  Edyta Krzymanska-Olejnik ,  Sergey Mamaev ,  Kenneth J. Rothschild

发明人： Jerzy Olejnik ,  Edyta Krzymanska-Olejnik ,  Sergey Mamaev ,  Kenneth J. Rothschild

IPC分类号： C07C245/00 ,  C07C291/00 ,  C12Q1/68 ,  C07K14/00 ,  G01N33/00 ,  C07H21/04

CPC分类号： C07H21/02 ,  C07H21/04 ,  G01N33/532 ,  Y02P20/55

摘要： The present invention relates to methods for the preparation of chemically aminoacylated tRNAs for the purpose of introduction of markers into nascent proteins. The present invention also relates to methods for the non-radioactive labeling, detection, quantitation and isolation of nascent proteins translated in a cellular or cell-free translation system utilizing chemically aminoacylated tRNAs. tRNA molecules are misaminoacylated with non-radioactive markers which may be non-native amino acids, amino acid analogs or derivatives. Markers may comprise cleavable moieties, detectable labels, reporter properties wherein markers incorporated into protein can be distinguished from unincorporated markers, or coupling agents which facilitate the detection and isolation of nascent protein from other components of the translation system.

摘要翻译： 本发明涉及用于制备化学氨基酰化tRNA以便将标记物引入新生蛋白质的方法。 本发明还涉及在使用化学氨酰化tRNA的细胞或无细胞翻译系统中翻译的新生蛋白的非放射性标记，检测，定量和分离的方法。 tRNA分子被非放射性标记物错氨基酰化，其可以是非天然氨基酸，氨基酸类似物或衍生物。 标记可以包含可切割部分，可检测标记，报告基因特性，其中掺入蛋白质的标记可以与非组合标记物或促进从翻译系统的其他组分检测和分离新生蛋白质的偶联剂区别。

公开(公告)号：US20080032417A1

公开(公告)日：2008-02-07

申请号：US11582294

申请日：2006-10-17

申请人： Jerzy Olejnik ,  Kenneth Rothschild

发明人： Jerzy Olejnik ,  Kenneth Rothschild

IPC分类号： C07D495/04 ,  C07K1/00 ,  G01N33/68

CPC分类号： G01N33/532 ,  C07D495/04 ,  C07K1/1077 ,  C07K1/13 ,  C07K1/14 ,  C07K1/16 ,  C07K1/22 ,  G01N2458/15 ,  Y10T436/24

摘要： This invention relates to agents and conjugates that can be used to detect and isolate target components from complex mixtures such as proteins and protein fragments from biological samples from in vivo and in vitro sources. Agents comprise a detectable group bound to a photoreactive group. Conjugates comprise agents coupled to substrates by covalent bounds which can be selectively cleaved with the administration of electromagnetic radiation. Targets substances labeled with detectable molecules can be easily identified and separated from a heterologous mixture of substances. Exposure of the conjugate to radiation releases the target in a functional form and completely unaltered. Using photocleavable molecular precursors as the conjugates, label can be incorporated into macromolecules, the nascent macromolecules isolated and the label completely removed. The invention also relates to targets isolated with these conjugates which may be useful as pharmaceutical agents or compositions that can be administered to humans and other mammals.

摘要翻译： 本发明涉及可用于从体内和体外来源的生物样品的复杂混合物如蛋白质和蛋白质片段检测和分离靶组分的试剂和缀合物。 试剂包含与光反应性基团结合的可检测基团。 缀合物包括通过共价界限偶联到底物的试剂，其可以通过施用电磁辐射被选择性地切割。 目标可以用可检测分子标记的物质可以很容易地识别，并与物质的异源混合物分离。 缀合物暴露于放射线以功能形式释放靶，完全不变。 使用可光致发光的分子前体作为共轭物，标记可以并入大分子中，分离出新生大分子，并将标记完全除去。 本发明还涉及用这些缀合物分离的靶，其可用作可以施用于人和其它哺乳动物的药剂或组合物。

公开(公告)号：US07252932B1

公开(公告)日：2007-08-07

申请号：US10049332

申请日：2000-08-23

申请人： Kenneth J. Rothschild ,  Sadanand Gite ,  Jerzy Olejnik ,  Sergey Mamaev ,  Mark Lim

发明人： Kenneth J. Rothschild ,  Sadanand Gite ,  Jerzy Olejnik ,  Sergey Mamaev ,  Mark Lim

IPC分类号： C12Q1/68

CPC分类号： G01N33/6803 ,  C12P21/02

摘要： This invention relates to non-radioactive markers that facilitate the detection and analysis of nascent proteins translated within cellular or cell-free translation systems. Nascent proteins containing these markers can be rapidly and efficiently detected, isolated and analyzed without the handling and disposal problems associated with radioactive reagents. Preferred markers are dipyrrometheneboron difluoride (4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) dyes.

摘要翻译： 本发明涉及促进在细胞或无细胞翻译系统内翻译的新生蛋白的检测和分析的非放射性标记物。 可以快速有效地检测，分离和分析含有这些标记物的蛋白质，而不会与放射性试剂相关的处理和处置问题。 优选的标记是二吡咯亚甲基二氟化物（4,4-二氟-4-硼酸-3a，4a-二氮杂 - 顺 - 并苯并三嗪）染料。