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Researchers have developed a miniaturized method for isolating lysosomes from cells that requires only 500,000 cells instead of tens of millions. The technique uses detergent-based destabilization of the plasma membrane followed by mechanical homogenization, combined with either magnetic nanoparticles or immunoprecipitation for lysosome enrichment. This approach not only drastically reduces required cell numbers but also produces higher purity samples and better proteome characterization compared to conventional methods.
Why it matters
This advancement enables lysosome research using rare cell types or limited clinical samples that were previously difficult to study due to high cell number requirements. The improved efficiency and purity could accelerate research into lysosomal storage diseases and other conditions involving lysosomal dysfunction.
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⚠️ Preprint – Noch nicht peer-reviewed
Dieser Artikel wurde noch nicht von unabhängigen Experten begutachtet. Die Ergebnisse sind vorläufig und sollten mit Vorsicht interpretiert werden.
Organelle enrichment presents a prerequisite for the unbiased biochemical analysis of cellular subcellular compartments. The performance of this step is decisive for the experiment’s success, and despite the high sensitivity of downstream analytical strategies, such as enzymatic assays, western blotting, or mass spectrometry (MS)-based OMICs approaches, typically tens of millions of cells are required, standing in stark contrast. Here, we demonstrate that selective rupture of the plasma membrane constitutes a limiting factor for the reduction of cell numbers and present an approach to overcome this limitation through detergent-based plasma membrane destabilization, followed by mechanical homogenization. By combination of this strategy with two common methods for lysosome enrichment, namely superparamagnetic iron oxide nanoparticles (SPIONs) and immunoprecipitation via 3xHA-tagged TMEM192 (TMEM IP), we scale down lysosome enrichment to only half a million cells and demonstrate that reduced input cell numbers yield superior results with respect to sample purity and organelle proteome characterization.
Source: Miniaturized Lysosome Enrichment through Selective Plasma Membrane Destabilization