Pezzi Hannah M, Niles David J, Schehr Jennifer L, Beebe David J, Lang Joshua M
Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, 1111 Highland Avenue, Madison, Wisconsin 53705, United States.
Department of Medicine and Carbone Cancer Center, University of Wisconsin-Madison, Madison, Wisconsin 53705, United States.
ACS Omega. 2018 Apr 30;3(4):3908-3917. doi: 10.1021/acsomega.7b01427. Epub 2018 Apr 6.
Magnetic bead-based analyte capture has emerged as a ubiquitous method in cell isolation, enabling the highly specific capture of target populations through simple magnetic manipulation. To date, no "one-size fits all" magnetic bead has been widely adopted leading to an overwhelming number of commercial beads. Ultimately, the ideal bead is one that not only facilitates cell isolation but also proves compatible with the widest range of downstream applications and analytic endpoints. Despite the diverse offering of sizes, coatings, and conjugation chemistries, few studies exist to benchmark the performance characteristics of different commercially available beads; importantly, these bead characteristics ultimately determine the ability of a bead to integrate into the user's assay. In this report, we evaluate bead-based cell isolation considerations, approaches, and results across a subset of commercially available magnetic beads (Dynabeads FlowComps, Dynabeads CELLection, GE Healthcare Sera-Mag SpeedBeads streptavidin-blocked magnetic particles, Dynabeads M-270s, Dynabeads M-280s) to compare and contrast both capture-specific traits (i.e., purity, capture efficacy, and contaminant isolations) and endpoint compatibility (i.e., protein localization, fluorescence imaging, and nucleic acid extraction). We identify specific advantages and contexts of use in which distinct bead products may facilitate experimental goals and integrate into downstream applications.
基于磁珠的分析物捕获已成为细胞分离中一种普遍使用的方法,通过简单的磁性操作就能高度特异性地捕获目标细胞群体。到目前为止,还没有一种“通用型”磁珠被广泛采用,这导致市面上的商业磁珠数量众多。最终,理想的磁珠不仅要便于细胞分离,还要与最广泛的下游应用和分析终点兼容。尽管磁珠在尺寸、涂层和偶联化学方面有多种选择,但很少有研究对不同市售磁珠的性能特征进行基准测试;重要的是,这些磁珠特性最终决定了磁珠融入用户检测方法的能力。在本报告中,我们评估了一系列市售磁珠(Dynabeads FlowComps、Dynabeads CELLection、通用电气医疗集团的Sera-Mag SpeedBeads链霉亲和素封闭磁性颗粒、Dynabeads M-270s、Dynabeads M-280s)基于磁珠的细胞分离考量因素、方法和结果,以比较和对比捕获特异性特征(即纯度、捕获效率和污染物分离)以及终点兼容性(即蛋白质定位、荧光成像和核酸提取)。我们确定了不同磁珠产品在促进实验目标和融入下游应用方面可能具有的特定优势和适用场景。