Presentation Information
[P04-529]Uncovering the relationship between Laurdan’s lifetime and the lipid membrane’s viscosity via TRES
○Zachary Nicolella1, Nozomi Morishia Watanabe1, Yukihiro Okamoto1, Hiroshi Umakoshi1 (1. University of Osaka Graduate School of Engineering Science (Japan))
Keywords:
Lipid Membranes,LNP,TRES,Viscosity
[Introduction]
Critical evaluation of the biophysical properties of lipid nanoparticles (LNPs) is one the most important steps in determining the efficency and stability of LNP based therapeutics. However, current classification relies on high through-put, low resultion techniques such as DLS. Issues such as probe mismatch, various probes reporting the same information via a different value, and long measurment times prevent the usage of more advnaced measurement techniques. Despite this, Laurdan—perhaps the most ubiquitous membrane probe—can, via a novel interpretation of its excitation mechanism, resolve these problems and provide a unified description of the interfacial region via one measurement.
[Method]
The decay assoicated spectrum (DAS) of the time-resolved emission sprectrum (TRES) of an extensive solvent library, Laurdan in various liposomes, and Laurdan under different depth-biased quencher were used along comparaitive anaylsis with of the time-resolved anisotropy of TMA-DPH.
[Results]
The TRES spectrum of Laurdan in various solvents showed no quantifiable relationship between Laurdan’s lifetime and neat solvent properties such as viscosity, dielectric constant, or quantum yield. Laurdan also showed no resolvable peak. Position shift in the the time-resolved area normalized emission spectra (TRANES) in solvent with the emission lineshape being established in the ealiest resolvable time window. Nonetheless, Laurdan, as expected, exhibited a classical solvatochromic shift in emission peak. Relationships between the lifetime of Laurdan and other lipid membrane probes were then examined in which a strong relationship with the appearnt micro-viscosity, as measured by TMA-DPH was found (R2=0.89, nRMSE=0.088). Laurdan’s emission in DOPC quenched by different depth biased quenchers also show that Laurdans emission undergoes a sequential spectral evolution.
[Conclusion]
Laurdan’s lifetime exhibits a relationship with membrane viscosity, with its solvatochromic properties and ease of measurement, Laurdan becomes a much more powerful probe, specifically for LNP characterization where high throughput and characterization of stability related bio-physcial properties are critical to a viable product.
Critical evaluation of the biophysical properties of lipid nanoparticles (LNPs) is one the most important steps in determining the efficency and stability of LNP based therapeutics. However, current classification relies on high through-put, low resultion techniques such as DLS. Issues such as probe mismatch, various probes reporting the same information via a different value, and long measurment times prevent the usage of more advnaced measurement techniques. Despite this, Laurdan—perhaps the most ubiquitous membrane probe—can, via a novel interpretation of its excitation mechanism, resolve these problems and provide a unified description of the interfacial region via one measurement.
[Method]
The decay assoicated spectrum (DAS) of the time-resolved emission sprectrum (TRES) of an extensive solvent library, Laurdan in various liposomes, and Laurdan under different depth-biased quencher were used along comparaitive anaylsis with of the time-resolved anisotropy of TMA-DPH.
[Results]
The TRES spectrum of Laurdan in various solvents showed no quantifiable relationship between Laurdan’s lifetime and neat solvent properties such as viscosity, dielectric constant, or quantum yield. Laurdan also showed no resolvable peak. Position shift in the the time-resolved area normalized emission spectra (TRANES) in solvent with the emission lineshape being established in the ealiest resolvable time window. Nonetheless, Laurdan, as expected, exhibited a classical solvatochromic shift in emission peak. Relationships between the lifetime of Laurdan and other lipid membrane probes were then examined in which a strong relationship with the appearnt micro-viscosity, as measured by TMA-DPH was found (R2=0.89, nRMSE=0.088). Laurdan’s emission in DOPC quenched by different depth biased quenchers also show that Laurdans emission undergoes a sequential spectral evolution.
[Conclusion]
Laurdan’s lifetime exhibits a relationship with membrane viscosity, with its solvatochromic properties and ease of measurement, Laurdan becomes a much more powerful probe, specifically for LNP characterization where high throughput and characterization of stability related bio-physcial properties are critical to a viable product.
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