一种适合针刺活检样品的蛋白质组学分析方法

针刺活检样品是一种重要的临床组织样品，其蕴涵的蛋白质信息，对了解人类疾病极为重要.然而，由于该样品的体积极小，其研究受到很大限制.本文建立和优化了适合针刺活检样品的基于双向电泳的蛋白质组学分析平台：通过直接抽提获得针刺活检组织的蛋白质样品；用24 cm 固定梯度干胶条(pH 3-10NL)等电聚焦及12.5% SDS-PAGE分离获得蛋白质样品；感兴趣的蛋白质点经过胰酶酶解后用MALDI TOF/TOF质谱分析.运用所建立的平台对3例来自3只不同大鼠的肝脏针刺活检样品进行分析，获得了多于2500个蛋白质点的高重复性的二维凝胶银染图谱.应用该方法分析人前列腺针刺活检样品的蛋白质组，同样获得了高质量、高重复性的结果.其中随机选取的包括低丰度点在内的57个蛋白质点，经胰酶酶解后进行MALDI串联质谱分析，均获得了高确定性的鉴定结果.通过建立针刺活检样品的蛋白质组学分析方法，为研究人类疾病的分子机制提供了必要的前提保障.

A Proteomic Method For Core Needle Biopsy Sample Characterization

Proteomic analysis of core needle biopsy (CNB) sample from patient populations is critical to our understanding of human disease,but has been hindered by its particular small size.Here,we present a method for the proteomic analysis of CNB sample based on the two dimensional electrophoresis.Proteins were extracted directly from 3 rat liver CNB specimens and a human prostate CNB sample.respectively.24 cm Immobiline DryStrip (pH 3-10NL) and 12.5% SDS-PAGE were introduced to separate the proteins.Interesting spots were analyzed by MALDI TOF/TOF mass spectrometry after tryptic digestion.With this method,consistent electrophoretic patterns of more than 2 500 protein spots were reproducibly obtained after silver staining,from rat liver CNB specimens.Qualitatively and quantitatively reproducible results also yield when the method was applied to a human prostate CNB sample.57 stochastically selected protein spots were analyzed by MALDI TOF/TOF moss spectrometry.and were identified with high confidence including faint ones.This simple and reproducible approach raises the opportunity of defining key molecular events of human disease pathologies.