A Lipid Mixing Assay to Accurately Quantify the Fusion of Outer Membrane Vesicles

Publication date: Available online 17 November 2019Source: MethodsAuthor(s): Yehou M. D. Gnopo, David PutnamAbstractThe intermixing of phospholipids from opposing bilayers, or membrane fusion, is a naturally occurring process that can be leveraged to produce hybrid vesicle systems. Optimizing the production of these hybrid vesicles requires accurate, sensitive and quantitative methods of the lipid mixing that occurs during fusion. A fluorescence-based assay that uses octadecyl-rhodamine B chloride to measure lipid mixing, or R18 assay, was developed by Hoekstra to investigate viral entry almost four decades ago. However, the R18 assay has so far only been used to measure heterotypic fusion events. Consequently, the fusion efficiencies that are calculated from the R18 assay underestimate the total number of fusion events and the true efficiency of vesicle fusions. In this article, we outline the experimental format and data analysis that is necessary to perform the R18 fusion assay and to accurately and reliably measure the true total fusion efficiency of outer membrane vesicles isolated from the Nissle 1917 strain of E. coli.
Source: Methods - Category: Molecular Biology Source Type: research

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Source: Colloids and Surfaces B: Biointerfaces - Category: Biochemistry Source Type: research
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Source: Colloids and Surfaces B: Biointerfaces - Category: Biochemistry Source Type: research
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Source: Oncology Letters - Category: Cancer & Oncology Tags: Oncol Lett Source Type: research
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Source: Oncology Letters - Category: Cancer & Oncology Tags: Oncol Lett Source Type: research
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Source: Journal of Molecular Structure - Category: Molecular Biology Source Type: research
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