CAJ | 학술논문

数据非依赖性采集( DIA)是随着定量蛋白质组学而建立的质谱扫描技术。 DIA能够获得扫描范围内所有母离子及二级子离子信息,不会造成低丰度离子信息的丢失,同时突破了高分辨质谱二级定量的通量限制。本研究基于静电场轨道阱Q-qIT-OT三合一质谱,发展了经典DIA方法以及WiSIM-DIA和Full MS-DIA两种全新DIA方法,并对Hela细胞全蛋白中添加的10条低浓度肽段进行定量分析,考察方法的线性、重现性和灵敏度。结果表明,3种方法的定量限均低至amol (14~435 amol),并展示出良好的线性和定性确证可靠性。其中,WiSIM-DIA基于超高分辨一级监测定量,与经典DIA优势互补；Full MS-DIA的选择窗口仅3 amu,能够直接进行搜库鉴定,实现了数据依赖性采集( DDA)和DIA的统一,摆脱了DIA依赖于DDA建立谱图库的局限性。
수거비의뢰성채집( DIA)시수착정량단백질조학이건립적질보소묘기술。 DIA능구획득소묘범위내소유모리자급이급자리자신식,불회조성저봉도리자신식적주실,동시돌파료고분변질보이급정량적통량한제。본연구기우정전장궤도정Q-qIT-OT삼합일질보,발전료경전DIA방법이급WiSIM-DIA화Full MS-DIA량충전신DIA방법,병대Hela세포전단백중첨가적10조저농도태단진행정량분석,고찰방법적선성、중현성화령민도。결과표명,3충방법적정량한균저지amol (14~435 amol),병전시출량호적선성화정성학증가고성。기중,WiSIM-DIA기우초고분변일급감측정량,여경전DIA우세호보；Full MS-DIA적선택창구부3 amu,능구직접진행수고감정,실현료수거의뢰성채집( DDA)화DIA적통일,파탈료DIA의뢰우DDA건립보도고적국한성。
Data independent acquisition ( DIA ) is a novel MS scan mode for quantitative proteomics, acquiring all precursors as well as fragments without any loss of low abundant ions, and breaks the throughput limitation of product ion quantification by high-resolution MS. Here we developed three DIA methods on quadrupole-linear ion trap-Orbitrap ( Q-qIT-OT ) Tribrid MS, classic DIA, as well as novel wide isolation window SIM scan ( WiSIM)-DIA and full scan-DIA ( Full MS-DIA) . Quantitative analysis of 10 low abundant peptides spiked in Hela cell digest was performed by the three methods for linearity, reproducibility and sensitivity evaluation. The results showed that the LOQs reached amol level ( 14-435 amol ) with good linearity and effective MS/MS confirmation. WiSIM-DIA utilizes ultra-high resolution SIM scan for quantification complementary with classic DIA. The isolation window of Full MS-DIA was down to 3 amu, and the data could be directly used for database searching, thus realizing the integration of data dependent acquisition ( DDA) and DIA, and avoiding the limitation of using spectra library.