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

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.