蛋白質(zhì)組學系列講座04基于質(zhì)譜的蛋白質(zhì)組學定量方法

基于質(zhì)譜的蛋白質(zhì)組學定量方法解

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長0102030405質(zhì)譜定量蛋白質(zhì)組學簡介標記定量法非標記定量法DIA技術靶向定量法CONTENTS課

程

目

錄質(zhì)譜定量蛋白質(zhì)組學簡介01解

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長蛋白質(zhì)組學（英語：proteomics），是以蛋白質(zhì)組為研究對象，研究細胞、組織或生物體蛋白質(zhì)組成及其變化規(guī)律的科學。Pictures

were

all

from

Bing.傳統(tǒng)Western

Blot法基于質(zhì)譜的蛋白質(zhì)組學方法蛋白有限抗體要求高高通量需求成千上萬種蛋白能篩選，能驗證高分辨，更準確解

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長1.2

質(zhì)譜儀工作原理Orbitrap

質(zhì)譜儀工作原理圖解

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質(zhì)譜儀工作示例解

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質(zhì)譜數(shù)據(jù)構成解

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牌Zoom

in解

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牌1.3

質(zhì)譜定量法主要有哪些？利用化學分子標簽的信號相應進行定量。利用峰面積進行定量。不依賴于母離子的信號相應，對一級和二級離子進行全掃描。對目標肽段進行精確定量。標記定量非標記定量靶向定量DIA新技術標記定量02解

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牌1.3

標記定量法的原理兩大主流標記定量方法：優(yōu)點：定量準確，穩(wěn)定性高，重現(xiàn)性好。缺點：大樣本量成本高基于報告離子的定量方法。知

識

金

牌1.4

標記定量法的實驗流程1234567解

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長相關文獻 知

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牌非標記定量03解

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牌3.1

非標記定量法的原理

顧名思義，非標記定量法是指不使用同位素標記物，而是直接根據(jù)質(zhì)譜峰的強度來對肽段或蛋白進行相對定量的方法，

文獻中一般稱為“l(fā)abel

free”。其定量原理是計算一級質(zhì)譜中每個肽段的信號強度在LC-MS

色譜上的積分面積，利用MaxQuant等軟件解析這些肽段的定量信息，從而獲得對應蛋白質(zhì)的相對定量值。此外，也可以通過計算譜圖數(shù)目進行相對定量。傳統(tǒng)定量方法：優(yōu)點：成本低，無樣本數(shù)限制，操作簡便缺點：重現(xiàn)性差解

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牌3.2

非標記定量法的流程解

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長4.1

DIA技術原理 知

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牌DIA（Data

Independent

Acquisition）是隨著定量蛋白質(zhì)組學而建立的質(zhì)譜掃描技術。DIA技術能夠獲得掃描范圍內(nèi)所有母離子和二級離子的信息，不會造成低豐度離子信息的丟失。Data

Dependent

Acquisition Data

Independent

Acquisition解

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牌DDADIA4.1

DIA技術原理Picturewas

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matearials

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AB

SCIEXtraining.解

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DIA實驗流程Picturewas

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靶向定量的原理 知

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牌PRM：平行反應監(jiān)測（Parallel

Reaction

Monitoring,

PRM）技術是一種基于目標肽段質(zhì)譜信號獲取，以此來進行蛋白質(zhì)定量的技術。解

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牌5.2

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