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

基于質(zhì)譜的蛋白質(zhì)組學(xué)定量方法解

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

程

目

錄質(zhì)譜定量蛋白質(zhì)組學(xué)簡介01解

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

were

all

from

Bing.傳統(tǒng)Western

Blot法基于質(zhì)譜的蛋白質(zhì)組學(xué)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(yàn)證高分辨，更準(zhǔn)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(gòu)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(xué)分子標(biāo)簽的信號(hào)相應(yīng)進(jìn)行定量。利用峰面積進(jìn)行定量。不依賴于母離子的信號(hào)相應(yīng)，對(duì)一級(jí)和二級(jí)離子進(jìn)行全掃描。對(duì)目標(biāo)肽段進(jìn)行精確定量。標(biāo)記定量非標(biāo)記定量靶向定量DIA新技術(shù)標(biāo)記定量02解

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牌1.3

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

識(shí)

金

牌1.4

標(biāo)記定量法的實(shí)驗(yàn)流程1234567解

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長統(tǒng)計(jì)分析生物信息學(xué)分析數(shù)據(jù)挖掘……Li,

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牌非標(biāo)記定量03解

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牌3.1

非標(biāo)記定量法的原理

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

文獻(xiàn)中一般稱為“l(fā)abel

free”。其定量原理是計(jì)算一級(jí)質(zhì)譜中每個(gè)肽段的信號(hào)強(qiáng)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(duì)應(yīng)蛋白質(zhì)的相對(duì)定量值。此外，也可以通過計(jì)算譜圖數(shù)目進(jìn)行相對(duì)定量。傳統(tǒng)定量方法：優(yōu)點(diǎn)：成本低，無樣本數(shù)限制，操作簡便缺點(diǎn)：重現(xiàn)性差解

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牌3.2

非標(biāo)記定量法的流程解

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長相關(guān)文獻(xiàn) 知

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DIA技術(shù)原理 知

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牌DIA（Data

Independent

Acquisition）是隨著定量蛋白質(zhì)組學(xué)而建立的質(zhì)譜掃描技術(shù)。DIA技術(shù)能夠獲得掃描范圍內(nèi)所有母離子和二級(jí)離子的信息，不會(huì)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(shù)原理Picturewas

modified

from

matearials

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AB

SCIEXtraining.解

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牌4.2

DIA實(shí)驗(yàn)流程Picturewas

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Tiannan

Guo,Nature

Communication,

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長相關(guān)文獻(xiàn) 知

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靶向定量的原理 知

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牌PRM：平行反應(yīng)監(jiān)測（Parallel

Reaction

Monitoring,

PRM）技術(shù)是一種基于目標(biāo)肽段質(zhì)譜信號(hào)獲取，以此來進(jìn)行蛋白質(zhì)定量的技術(shù)。解

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牌5.2

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