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METHODS FOR IDENTIFYING PROTEINS THAT BIND LIGANDS

机译:识别结合蛋白的方法

摘要

Provided herein are methods of identifying a protein capable of binding a ligand, the method comprising: (a) contacting the ligand with two or more samples comprising a plurality of proteins in a solution; (b) separating the proteins bound to the ligand ("bound proteins") from the proteins that are not bound to the ligand ("unbound proteins") in each sample; (c) denaturing and digesting the bound proteins to form a plurality of peptides in each sample; (d) quantifying a plurality of molecular features contained in the plurality of peptides in each sample, wherein the molecular features are defined as having a mass to charge ratio, retention time, and peak intensity as measured by mass spectrometry; and (e) ranking the molecular features that exhibit a statistically significant difference in quantity between the samples contacted with the ligand and a sample that is not contacted with the ligand ("statistically significant molecular feature").
机译:本文提供了鉴定能够结合配体的蛋白质的方法,该方法包括:(a)使配体与包含溶液中的多种蛋白质的两个或更多个样品接触; (b)从每个样品中分离与配体结合的蛋白质(“结合蛋白质”)与未与配体结合的蛋白质(“未结合蛋白质”); (c)使结合的蛋白质变性和消化,以在每个样品中形成多个肽; (d)定量每个样品的多个肽中包含的多个分子特征,其中所述分子特征被定义为具有通过质谱法测量的质荷比,保留时间和峰强度; (e)对与配体接触的样品和不与配体接触的样品之间在数量上具有统计学显着差异的分子特征进行分级(“统计学上显着的分子特征”)。

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