RamalloGuevara Carina, Paulssen Dorothea, Popova Anna A, Hopf Carsten, Levkin Pavel A
Center for Mass Spectrometry and Optical Spectroscopy (CeMOS), Mannheim University of Applied Sciences, Paul-Wittsack Str. 10, Mannheim, 68163, Germany.
Karlsruhe Institute of Technology (KIT), Institute of Biological and Chemical Systems - Functional Molecular Systems (IBCS-FMS), Hermann-von-Helmholtz-Platz 1, Eggenstein-Leopoldshafen, 76344, Germany.
Adv Biol (Weinh). 2021 Mar;5(3):e2000279. doi: 10.1002/adbi.202000279. Epub 2021 Feb 9.
In pharmaceutical research and development, cell-based assays are primarily used with readout that rely on fluorescence-based and other label-dependent techniques for analysis of different cellular processes. Superhydrophobic-hydrophilic droplet microarrays (DMA) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) have recently emerged as key technologies for miniaturized high-throughput cell assays and for label-free molecular high-content drug profiling, respectively. Here, nanoliter-scale cell assays are integrated on DMAs with MALDI-MS imaging (MALDI-MSI) approaches to a droplet microarray-mass spectrometry imaging (DMA-MSI) platform. Using A549 lung cancer cells, concentration-response profiling of a pharmaceutical compound, the fatty acid synthase inhibitor GSK2194069, are demonstrated. Direct cell culture on DMAs enables combination of microscopy and high speed, high molecular content analysis using MALDI-MSI. Miniaturization of array spots down to 0.5 mm confining 40 nL droplets allows for MALDI imaging analysis of as few as ten cells per spot. Partial automation ensures a fast sample preparation workflow. Taken together, the integrated DMA-MSI platform that combines MALDI-MSI, as a label-free analytical readout, with the miniaturized droplet microarray platform is a valuable complement to high throughput cell-based assays technologies.
在药物研发中，基于细胞的检测主要用于依赖荧光及其他标记依赖技术的读数，以分析不同的细胞过程。超疏水-亲水液滴微阵列（DMA）和基质辅助激光解吸/电离（MALDI）质谱（MS）最近分别成为用于小型化高通量细胞检测和无标记分子高内涵药物分析的关键技术。在此，纳升规模的细胞检测通过MALDI-MS成像（MALDI-MSI）方法集成到DMA上，形成了液滴微阵列-质谱成像（DMA-MSI）平台。使用A549肺癌细胞，展示了一种药物化合物——脂肪酸合酶抑制剂GSK2194069的浓度-反应分析。在DMA上直接进行细胞培养，可将显微镜检查与使用MALDI-MSI的高速、高分子含量分析相结合。将阵列点小型化至0.5毫米，容纳40纳升液滴，使得每个点少至十个细胞也能进行MALDI成像分析。部分自动化确保了快速的样品制备流程。综上所述，将作为无标记分析读数的MALDI-MSI与小型化液滴微阵列平台相结合的集成DMA-MSI平台，是基于细胞的高通量检测技术的宝贵补充。
