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    Method and apparatus for the identification and quantification of biomolecules
    1.
    发明授权
    Method and apparatus for the identification and quantification of biomolecules 有权
    用于鉴定和定量生物分子的方法和装置

    公开(公告)号：US07166441B2

    公开(公告)日：2007-01-23

    申请号：US10327342

    申请日：2002-12-20

    申请人： Timothy K. Nadler ,  Kenneth G. Parker ,  George J. Vella ,  Barrie G. Wagenfeld ,  Yulin Huang ,  Robert J. Lotti

    发明人： Timothy K. Nadler ,  Kenneth G. Parker ,  George J. Vella ,  Barrie G. Wagenfeld ,  Yulin Huang ,  Robert J. Lotti

    IPC分类号： C12Q1/37

    CPC分类号： G01N33/6848 ,  G01N33/6851

    摘要： Polypeptides are electroblotted through a digestion membrane to a composite capture membrane that can be directly analyzed using mass spectrometry. The molecular weights of the fragments generated by the digestion membrane are then used to identify the polypeptide from which they originated. The digestion membrane contains an immobilized protease such as trypsin, which cleaves the electroblotted polypeptides into fragments during electroblotting with such high enzyme cleavage capacity and efficiency that one pass of the polypeptide through the membrane is sufficient. The peptide fragments are collected onto a composite capture membrane that is chemically treated, for example by adding a mixture of nitrocellulose and MALDI matrix, so as to absorb peptides near the surface to facilitate desportion, thereby increasing the sensitivity of subsequent analysis by MALDI-TOF mass spectrometry. A wide variety of application are disclosed including identifying proteins separated on a gel or within a tissue sample.

    摘要翻译： 多肽通过消化膜电渗透到复合捕获膜，可以使用质谱直接分析。 然后将消化膜产生的片段的分子量用于鉴定它们起源的多肽。 消化膜含有固定化的蛋白酶，例如胰蛋白酶，其在电印迹期间将电印迹多肽切割成片段，其具有如此高的酶切割能力和多肽通过膜的一次通过足够的效率。 将肽片段收集到化学处理的复合捕获膜上，例如通过加入硝酸纤维素和MALDI基质的混合物，以吸收表面附近的肽以促进分离，从而提高随后的MALDI-TOF分析的灵敏度 质谱。 公开了各种各样的应用，包括鉴定在凝胶上或组织样品中分离的蛋白质。

    Process for analyzing protein samples
    2.
    发明授权
    Process for analyzing protein samples 有权
    蛋白质样品分析方法

    公开(公告)号：US07045296B2

    公开(公告)日：2006-05-16

    申请号：US09851058

    申请日：2001-05-08

    申请人： Kenneth C. Parker ,  Timothy K. Nadler ,  George J. Vella ,  Yulin Huang ,  Rudolf H. Aebersold ,  Marcus B. Smolka

    发明人： Kenneth C. Parker ,  Timothy K. Nadler ,  George J. Vella ,  Yulin Huang ,  Rudolf H. Aebersold ,  Marcus B. Smolka

    IPC分类号： G01N33/53 ,  G01N24/00

    CPC分类号： G01N33/6803 ,  Y10S435/964 ,  Y10T436/24 ,  Y10T436/25125

    摘要： Methods using gel electrophoresis and mass spectrometry for the rapid, quantitative analysis of proteins or protein function in mixtures of proteins derived from two or more samples in one unit operation are disclosed. In one embodiment the method includes (a) preparing an extract of proteins from each of at least two different samples; (b) providing a set of substantially chemically identical and differentially isotopically labeled protein reagents; (c) reacting the extract of proteins from different samples of step (a) with a different isotopically labeled reagent from the set of step (b) to provide two or more sets of isotopically differentially labeled proteins; (d) mixing each of the two or more sets of isotopically labeled proteins to form a single mixture of isotopically differentially labeled proteins; (e) electrophoresing the mixture of step (d) by an electrophoresing method capable of separating proteins within the mixture; (f) digesting at least a portion of one or more separated proteins of step (e) and (g) detecting the difference in the expression levels of the proteins in the two samples by mass spectrometry based on one or more peptides in the sample of labeled peptides. The analytical method can be used for qualitative and particularly for quantitative analysis of global protein expression profiles in cells and tissues, i.e. the quantitative analysis of proteomes.

    摘要翻译： 公开了使用凝胶电泳和质谱法在一个单元操作中从两个或更多个样品衍生的蛋白质的蛋白质或蛋白质功能的快速，定量分析的方法。 在一个实施方案中，该方法包括（a）从至少两种不同样品中的每一种制备蛋白质提取物; （b）提供一组基本上化学相同和差异同位素标记的蛋白质试剂; （c）使来自步骤（a）的不同样品的蛋白质提取物与来自步骤（b）的组的不同的同位素标记的试剂反应，以提供两组或更多组同位素差异标记的蛋白质; （d）混合两组或更多组同位素标记的蛋白质以形成同位素差异标记的蛋白质的单一混合物; （e）通过能够分离混合物内的蛋白质的电泳方法电泳步骤（d）的混合物; （f）消化步骤（e）中的一种或多种分离的蛋白质的至少一部分，和（g）基于样品中的一种或多种肽通过质谱法检测两种样品中的蛋白质的表达水平的差异 标记肽。 分析方法可用于定性，特别是定量分析细胞和组织中的全球蛋白表达谱，即蛋白质组的定量分析。

    Fluid processing device
    4.
    发明授权
    Fluid processing device 失效
    流体处理装置

    公开(公告)号：US07491363B2

    公开(公告)日：2009-02-17

    申请号：US10891646

    申请日：2004-07-15

    申请人： Timothy K. Nadler

    发明人： Timothy K. Nadler

    IPC分类号： G01N35/00

    CPC分类号： B01L3/50273 ,  B01L3/5025 ,  B01L3/502738 ,  B01L2200/0621 ,  B01L2300/0816 ,  B01L2300/087 ,  B01L2400/0409 ,  B01L2400/086

    摘要： A fluid processing device is provided that enables the controlled flow of a liquid sample along a fluid processing pathway through various sample-containment regions and is free of fluid flow blockages or valves along the processing pathway. A system is also provided for processing the device and includes a rotatable platen and alignment features that can hold the fluid processing device in two or more different orientations on the rotatable platen. A method is also provided for processing a sample, in the device, with the system.

    摘要翻译： 提供了一种流体处理装置，其使液体样品沿着流体处理途径受到不同的样品容纳区域的受控流动，并且沿着处理途径没有流体流动阻塞或阀。 还提供了一种用于处理该装置的系统，并且包括可旋转的压板和可将流体处理装置保持在可旋转压盘上的两个或多个不同取向的对准特征。 还提供了一种用于在设备中与系统一起处理样品的方法。

    Protein imaging
    5.
    发明授权
    Protein imaging 失效
    蛋白成像

    公开(公告)号：US5756717A

    公开(公告)日：1998-05-26

    申请号：US448822

    申请日：1995-05-24

    申请人： Sandeep K. Paliwal ,  Timothy K. Nadler ,  Laszlo Varady ,  Fred E. Regnier

    发明人： Sandeep K. Paliwal ,  Timothy K. Nadler ,  Laszlo Varady ,  Fred E. Regnier

    IPC分类号： B01J20/32 ,  C07K1/22 ,  C08B37/00 ,  C07K1/16 ,  C07K1/36

    CPC分类号： B01J20/26 ,  B01J20/268 ,  B01J20/285 ,  B01J20/291 ,  C07K1/22 ,  B01D15/3852

    摘要： Disclosed are specifically designed binding, "protein imaged" sorbents which reversibly bind with high specificity and affinity a preselected macromolecule, specifically a protein. The sorbents define one or more cavities which have a binding surface complementary in shape to the molecular surface of the macromolecule and a plurality of positively and negatively charged chemical moieties spatially distributed in a mirror image and charge inverse of a subset of the ionizable groups on the molecular surface of the macromolecule. Also disclosed are methods of producing such sorbents, useful over a range of conditions, for both preparative and analytical chromatographic separations or for use in various types of analyses.

    摘要翻译： 公开了特异性设计的结合，“蛋白质成像”吸附剂，其以特异性和亲和力以预先选择的大分子，特别是蛋白质可逆地结合。 吸附剂限定一个或多个空腔，其具有与大分子的分子表面形状互补的结合表面，以及空间上分布在镜像中的多个带正电荷和带负电荷的化学部分，并且电离反应 分子表面的大分子。 还公开了制备在一系列条件下用于制备和分析色谱分离或用于各种类型分析的这种吸附剂的方法。