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FULL-MDS: Fluorescent Universal Lipid Labeling for Microfluidic Diffusional Sizing

[Image: see text] Microfluidic diffusional sizing (MDS) is a recent and powerful method for determining the hydrodynamic sizes and interactions of biomolecules and nanoparticles. A major benefit of MDS is that it can report the size of a fluorescently labeled target even in mixtures with complex, un...

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Detalles Bibliográficos
Autores principales: Baron, Jasmin, Bauernhofer, Lena, Devenish, Sean R. A., Fiedler, Sebastian, Ilsley, Alison, Riedl, Sabrina, Zweytick, Dagmar, Glueck, David, Pessentheiner, Ariane, Durand, Grégory, Keller, Sandro
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9850350/
https://www.ncbi.nlm.nih.gov/pubmed/36574263
http://dx.doi.org/10.1021/acs.analchem.2c03168
Descripción
Sumario:[Image: see text] Microfluidic diffusional sizing (MDS) is a recent and powerful method for determining the hydrodynamic sizes and interactions of biomolecules and nanoparticles. A major benefit of MDS is that it can report the size of a fluorescently labeled target even in mixtures with complex, unpurified samples. However, a limitation of MDS is that the target itself has to be purified and covalently labeled with a fluorescent dye. Such covalent labeling is not suitable for crude extracts such as native nanodiscs directly obtained from cellular membranes. In this study, we introduce fluorescent universal lipid labeling for MDS (FULL-MDS) as a sparse, noncovalent labeling method for determining particle size. We first demonstrate that the inexpensive and well-characterized fluorophore, Nile blue, spontaneously partitions into lipid nanoparticles without disrupting their structure. We then highlight the key advantage of FULL-MDS by showing that it yields robust size information on lipid nanoparticles in crude cell extracts that are not amenable to other sizing methods. Furthermore, even for synthetic nanodiscs, FULL-MDS is faster, cheaper, and simpler than existing labeling schemes.