Protein biomarkers on tissue as imaged via MALDI mass spectrometry: A systematic approach to study the limits of detection.
Protein biomarkers on tissue as imaged via MALDI mass spectrometry: A systematic approach to study the limits of detection.
复制标题
通过 MALDI 质谱成像的组织上的蛋白质生物标志物：研究检测限的系统方法。
DOI：
10.1002/pmic.201500515
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发表时间：
2016
期刊：
影响因子：
3.4
通讯作者：
Gambhir,SanjivS
中科院分区：
文献类型：
--
作者：
vandeVen,StephanieMWY;Bemis,KyleD;Lau,Kenneth;Adusumilli,Ravali;Kota,Uma;Stolowitz,Mark;Vitek,Olga;Mallick,Parag;Gambhir,SanjivS
MALDI质谱成像（MSI）正在成为跨组织切片的蛋白质和肽成像工具。尽管进行了广泛的研究，但目前还没有基线研究评估该技术检测组织切片中多种蛋白质的潜在能力。在这项研究中，我们从检测限、变异系数、空间分辨率和内源性组织蛋白的鉴定等方面开发了一种系统的方法来表征MALDI - MSI工作流程。我们的目标是量化这些不同蛋白质和肽的价值，以便更深入地了解使用该技术发现蛋白质生物标志物的可行性。对照蛋白和多肽以连续稀释的方式沉积在薄切片的小鼠异种移植组织上。在我们的实验装置中，组织切片的变异系数<30%，空间分辨率为200 μm（或更大）。组织中蛋白质和多肽的检出限在微摩尔到毫摩尔范围内。蛋白质鉴定只能对组织中存在高丰度的蛋白质进行。这些结果为MALDI - MSI在发现新的候选生物标志物方面的应用提供了基线，并为比较不同MALDI - MSI工作流程提供了新的基准策略。
MALDI mass spectrometry imaging (MSI) is emerging as a tool for protein and peptide imaging across tissue sections. Despite extensive study, there does not yet exist a baseline study evaluating the potential capabilities for this technique to detect diverse proteins in tissue sections. In this study, we developed a systematic approach for characterizing MALDI‐MSI workflows in terms of limits of detection, coefficients of variation, spatial resolution, and the identification of endogenous tissue proteins. Our goal was to quantify these figures of merit for a number of different proteins and peptides, in order to gain more insight in the feasibility of protein biomarker discovery efforts using this technique. Control proteins and peptides were deposited in serial dilutions on thinly sectioned mouse xenograft tissue. Using our experimental setup, coefficients of variation were <30% on tissue sections and spatial resolution was 200 μm (or greater). Limits of detection for proteins and peptides on tissue were in the micromolar to millimolar range. Protein identification was only possible for proteins present in high abundance in the tissue. These results provide a baseline for the application of MALDI‐MSI towards the discovery of new candidate biomarkers and a new benchmarking strategy that can be used for comparing diverse MALDI‐MSI workflows.