FULL-MDS: Fluorescent Universal Lipid Labeling for Microfluidic Diffusional Sizing
Jasmin Baron1,2,3, Lena Bauernhofer1,2,3, Sean R A Devenish4
1Biophysics, Institute of Molecular Biosciences (IMB), NAWI Graz, University of Graz, Humboldtstr. 50/III, Graz 8010, Austria.
Analytical Chemistry
|December 27, 2022
Summary
We developed fluorescent universal lipid labeling for microfluidic diffusional sizing (FULL-MDS), a new noncovalent method. FULL-MDS accurately sizes lipid nanoparticles in complex samples, overcoming limitations of current techniques.
Area of Science:
- Biotechnology
- Nanotechnology
- Analytical Chemistry
Background:
- Microfluidic diffusional sizing (MDS) is effective for determining biomolecule and nanoparticle size.
- Current MDS methods require target purification and covalent fluorescent labeling, unsuitable for crude samples like native nanodiscs.
Purpose of the Study:
- Introduce fluorescent universal lipid labeling for MDS (FULL-MDS) as a noncovalent labeling technique.
- Enable size determination of lipid nanoparticles in complex, unpurified biological extracts using MDS.
Main Methods:
- Utilized Nile blue, an inexpensive fluorophore, for noncovalent labeling of lipid nanoparticles.
- Applied FULL-MDS to assess the size of lipid nanoparticles in crude cell extracts.
- Compared FULL-MDS performance with existing labeling methods for synthetic nanodiscs.
Main Results:
- Nile blue spontaneously partitions into lipid nanoparticles without structural disruption.
- FULL-MDS successfully provided robust size information for lipid nanoparticles in crude cell extracts.
- FULL-MDS demonstrated to be faster, cheaper, and simpler than existing labeling schemes for synthetic nanodiscs.
Conclusions:
- FULL-MDS offers a versatile and efficient approach for sizing lipid nanoparticles, particularly in complex biological matrices.
- This noncovalent labeling method expands the applicability of MDS to previously inaccessible sample types.
- FULL-MDS presents a significant advancement for nanoparticle characterization in research and diagnostics.
