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    • 86. 发明授权
    • Detecting cells secreting a protein of interest
    • 检测分泌感兴趣的蛋白质的细胞
    • US09588117B2
    • 2017-03-07
    • US14583351
    • 2014-12-26
    • Berkeley Lights, Inc.
    • Kevin T. Chapman
    • G01N33/569G01N33/68
    • G01N33/56972G01N33/56911G01N33/56961G01N33/56966G01N33/6854G01N2458/00G01N2469/10
    • In some cases, the described systems and methods include obtaining a cell sample containing multiple antibody-producing cells. In such cases, the cells can be tagged with a cross-linking reagent having a first portion configured to bind to a marker on the antibody-producing cells and a second portion configured to bind to an antigen of interest. In some instances, the tagged antibody-producing cells are exposed to the antigen of interest such that the antigen becomes bound to the cells. In some such instances, the antibody-producing cells are also allowed to produce an antibody, such that a portion of the antibody-producing cells produce an antigen-specific antibody that binds to the antigen of interest. To identify cells that produce the antigen-specific antibody, the tagged cells can be exposed to a labeled secondary antibody that is configured to bind to the antigen-specific antibody. Other implementations are also described.
    • 在一些情况下,所描述的系统和方法包括获得含有多个产生抗体的细胞的细胞样品。 在这种情况下,可以用交联试剂对细胞进行标记,所述交联试剂具有构建成结合抗体产生细胞上的标记的第一部分和被配置成与感兴趣的抗原结合的第二部分。 在一些情况下,标记的产生抗体的细胞暴露于感兴趣的抗原,使得抗原与细胞结合。 在一些这样的情况下,也允许产生抗体的细胞产生抗体,使得产生一部分抗体的细胞产生与感兴趣的抗原结合的抗原特异性抗体。 为了鉴定产生抗原特异性抗体的细胞,标记的细胞可以暴露于被配置为结合抗原特异性抗体的标记的二抗。 还描述了其他实现。
    • 87. 发明授权
    • Immunoaffinity isolation of modified peptides from complex mixtures
    • 来自复杂混合物的修饰肽的免疫亲和分离
    • US09587013B2
    • 2017-03-07
    • US13868387
    • 2013-04-23
    • Cell Signaling Technology, Inc.
    • John RushHui ZhangXiangming ZhaMichael J CombYi Tan
    • G01N33/68C07K16/18B01D15/38C07K1/22C07K16/44G01N30/72G01N30/88
    • C07K16/18B01D15/3809B01D15/3828C07K1/22C07K16/44G01N30/7233G01N33/6842G01N33/6848G01N33/6854G01N2030/8831G01N2440/14
    • 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.
    • 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。