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　　在現(xiàn)在的蛋白質(zhì)組學(xué)研究熱潮中，很明顯，Evosep One的短梯度高通量與質(zhì)譜的DIA技術(shù)已成為一種成功的組合。

　　在本次Evosep舉辦的網(wǎng)絡(luò)研討會(huì)期間，弗洛里安·邁耶(Florian Meier， Friedrich Shiller University，Jena)和瓦迪姆·德米切夫(Vadim Demichev，克里克學(xué)院和劍橋大學(xué))將討論Evosep One如何與DIA技術(shù)碰撞出火花。

　　杭州紐藍(lán)誠(chéng)邀您參加此次Evosep One會(huì)議觀看。

　　會(huì)議時(shí)間：2021年5月27日晚10點(diǎn)-11點(diǎn)

　　講者及主題介紹：

　　主題：A PRACTICAL GUIDE TO HIGH-THROUGHPUT DATA-INDEPENDENT ACQUISITION

　　講者：Florian Meier

　　內(nèi)容：對(duì)于基于高通量質(zhì)譜的蛋白質(zhì)組學(xué)而言，DIA技術(shù)是一種越來(lái)越有吸引力的策略。 本演講將介紹最新的采集方法，例如非?？焖俚腛rbitrap質(zhì)量分析和diaPASEF。 研究者突出顯示了關(guān)鍵的方法參數(shù)，并說(shuō)明了它們對(duì)短梯度的優(yōu)化。 同時(shí)，研究者將討論離子遷移質(zhì)譜儀如何將其自身的特性如提高離子采樣效率，提高靈敏度并降低光譜復(fù)雜度的優(yōu)勢(shì)與Evosep結(jié)合起來(lái)，，從而實(shí)現(xiàn)每天跑60個(gè)樣品的高通量水準(zhǔn)，對(duì)6,000種蛋白質(zhì)進(jìn)行準(zhǔn)確的無(wú)標(biāo)記定量。

　　Data-independent acquisition (DIA) is an increasingly attractive strategy for high-throughput mass spectrometry-based proteomics. This presentation will give an introduction to state-of-the-art acquisition methods such as very fast Orbitrap mass analysis and diaPASEF.

　　We highlight key method parameters and illustrate their optimization for short gradients. Finally, we will discuss the integration of ion mobility spectrometry to improve ion sampling efficiency, increase sensitivity and reduce spectral complexity, leading to accurate label-free quantification of 6,000 proteins at a throughput of 60 samples per day.

　　主題：BEYOND 5000 PROTEINS IN 4.8 MINUTES FROM LOW SAMPLE AMOUNTS

　　講者：Vadim Demichev

　　內(nèi)容：在針對(duì)蛋白質(zhì)組學(xué)的研究中，對(duì)高通量的需求越來(lái)越迫切。在之前幾場(chǎng)的講座中，已經(jīng)介紹了DIA-NN，這是一個(gè)基于神經(jīng)網(wǎng)絡(luò)的軟件套件，能夠在60秒的時(shí)間內(nèi)獲取復(fù)雜DIA蛋白質(zhì)組的數(shù)據(jù)。 現(xiàn)在，研究者展示了DIA-NN可以應(yīng)用到timsTOF Pro和Evosep One上，以對(duì)200ng的Hela樣本為例，采取200樣本/天的檢測(cè)方法，定量分析得到5000多種蛋白質(zhì)。該工作流程可以實(shí)現(xiàn)高通量蛋白質(zhì)組學(xué)，其數(shù)據(jù)質(zhì)量和樣品量要求可與長(zhǎng)梯度nanoLC方法相媲美。

　　There is an increasing need for fast methods in proteomics. We have previously introduced DIA-NN, a neural network-based software suite capable of processing complex DIA proteomic data acquired with gradients as fast as 60-seconds. We now show that DIA-NN quantifies over 5000 proteins from just 200ng of a HeLa sample analysed with the 200 samples/day method on an Evosep One coupled to a timsTOF Pro.

　　This workflow enables high-throughput proteomics with data quality and sample amount requirements comparable to long-gradient nanoLC methods.