尺寸排除色谱与高通量电荷检测质谱联用分析大蛋白质和病毒样颗粒

IF 6.7 1区 化学 Q1 CHEMISTRY, ANALYTICAL
Raj A. Parikh, Lohra M. Miller, Benjamin E. Draper, Lavelay Kizekai, Balasubrahmanyam Addepalli, Michelle Chen, Matthew A. Lauber* and Martin F. Jarrold*, 
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引用次数: 0

摘要

电荷检测质谱(CD-MS)是一种新兴的单粒子技术,其中m/z和电荷都是单独测量的,以确定每个离子的质量。它特别适合于分析高质量和非均质样品。对于传统的质谱,高质量样品的电荷态分辨率的损失阻碍了质谱与尺寸排除色谱(SEC)等分离技术的直接耦合,并迫使使用低分辨率的检测器。在这里,我们展示了CD-MS如何利用高通量方法,如多离子电荷提取(MICE),可以匹配SEC的时间尺度,将适合SEC-CD-MS分离分析的样品扩展到兆道尔顿区甚至更高。作为这项工作的一部分,我们开发了低流量超宽孔(1000 Å孔径)SEC,使用窄孔柱来优化SEC与CD-MS之间的耦合。对单克隆抗体、甲状腺球蛋白、噬菌体Qβ病毒样颗粒(VLPs)和乙型肝炎病毒VLPs的分析,显示了SEC-CD-MS在宽质量范围内的能力,包括以前用ms无法在线分离的高质量范围。这些发现得到了利用多角度光散射(SEC-MALS)进行的平行研究的补充。SEC-CD-MS和SEC-MALS为复杂生物制剂和纳米颗粒的表征提供了有价值的补充信息。最后,我们的结果为高通量CD-MS与其他大分子和小分子分离技术的整合打开了大门。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Coupling of Size Exclusion Chromatography to High Throughput Charge Detection Mass Spectrometry for the Analysis of Large Proteins and Virus-like Particles

Charge detection mass spectrometry (CD-MS) is an emerging single-particle technique where both the m/z and charge are measured individually to determine each ion’s mass. It is particularly well-suited for analyzing high mass and heterogeneous samples. With conventional MS, the loss of charge state resolution with high mass samples has hindered the direct coupling of MS to separation techniques like size exclusion chromatography (SEC) and forced the use of lower resolution detectors. Here, we show how CD-MS, leveraging high-throughput methods such as multiple ion charge extraction (MICE), can match the time scale of SEC, extending the samples amenable to separation analysis by SEC-CD-MS into the megadalton regime and beyond. As part of this work, we have developed low flow ultrawidepore (1000 Å pore size) SEC using narrow bore columns to optimize the coupling between SEC and CD-MS. The analysis of monoclonal antibodies, thyroglobulin, bacteriophage Qβ virus-like particles (VLPs), and hepatitis B virus VLPs, showcases the capabilities of SEC-CD-MS over a broad mass range including the high mass range previously inaccessible for online separation with MS. These findings are complemented by a parallel study using multiangle light scattering (SEC-MALS). SEC-CD-MS and SEC-MALS provide complementary information that is valuable for characterization of complex biologics and nanoparticles. Finally, our results open the door to integration of high throughput CD-MS with other separation techniques for both large and small macromolecules.

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来源期刊
Analytical Chemistry
Analytical Chemistry 化学-分析化学
CiteScore
12.10
自引率
12.20%
发文量
1949
审稿时长
1.4 months
期刊介绍: Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.
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