Studies of phosphatidylcholine vesicles by spectroturbidimetric and dynamic light scattering methods

Khlebtsov, Boris N.; Kovler, L. A.; Богатырев, В. А.; Khlebtsov, Nikolai G.; Shchyogolev, S. Yu.

doi:10.1016/s0022-4073(02)00322-9

Cited by 14 publications

(8 citation statements)

References 19 publications

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“…We assumed in the library calculations an EV membrane thickness of 5 nm [ 49 , 50 ] and an EV lamellarity of 1 for biological membranes. We used for the medium and EV refractive indices the equations derived for water [ 51 ] and egg PC [ 52 ], respectively, as described previously [ 40 ]. We then calculated the turbidity spectra of F1 encompassing wells 51–111 ( Figure 5 d) by removing the background and multiplying the measured absorbance values by 2.303 (Equation (1), Figure 5 e) [ 53 ].…”

Section: Resultsmentioning

confidence: 99%

Development of Novel High-Resolution Size-Guided Turbidimetry-Enabled Particle Purification Liquid Chromatography (PPLC): Extracellular Vesicles and Membraneless Condensates in Focus

Kaddour

Lyu

Shouman

et al. 2020

IJMS

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Acellular particles (extracellular vesicles and membraneless condensates) have important research, drug discovery, and therapeutic implications. However, their isolation and retrieval have faced enormous challenges, impeding their use. Here, a novel size-guided particle purification liquid chromatography (PPLC) is integrated into a turbidimetry-enabled system for dye-free isolation, online characterization, and retrieval of intact acellular particles from biofluids. The chromatographic separation of particles from different biofluids—semen, blood, urine, milk, and cell culture supernatants—is achieved using a first-in-class gradient size exclusion column (gSEC). Purified particles are collected using a fraction collector. Online UV–Vis monitoring reveals biofluid-dependent particle spectral differences, with semen being the most complex. Turbidimetry provides the accurate physical characterization of seminal particle (Sp) lipid contents, sizes, and concentrations, validated by a nanoparticle tracking analysis, transmission electron microscopy, and naphthopyrene assay. Furthermore, different fractions of purified Sps contain distinct DNA, RNA species, and protein compositions. The integration of Sp physical and compositional properties identifies two archetypal membrane-encased seminal extracellular vesicles (SEV)—notably SEV large (SEVL), SEV small (SEVS), and a novel non-archetypal-membraneless Sps, herein named membraneless condensates (MCs). This study demonstrates a comprehensive yet affordable platform for isolating, collecting, and analyzing acellular particles to facilitate extracellular particle research and applications in drug delivery and therapeutics. Ongoing efforts focus on increased resolution by tailoring bead/column chemistry for each biofluid type.

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Section: Resultsmentioning

confidence: 99%

Development of Novel High-Resolution Size-Guided Turbidimetry-Enabled Particle Purification Liquid Chromatography (PPLC): Extracellular Vesicles and Membraneless Condensates in Focus

Kaddour

Lyu

Shouman

et al. 2020

IJMS

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“…9 B. We adopt the same strategy to measure the thickness of POPC membranes, using the wavelength-dependent refractive index for egg-phosphatidylcholine from Khlebtsov et al (53) and the area per lipid from Kucerka et al (54). The vesicle size measured by DLS for the POPC sample shown in Fig.…”

Section: Application 2 Measuring Membrane Thickness With Turbiditymentioning

confidence: 99%

Core-Shell Modeling of Light Scattering by Vesicles: Effect of Size, Contents, and Lamellarity

Wang

Miller

Szostak

2019

Biophysical Journal

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Having a fast, reliable method for characterizing vesicles is vital for their use as model cell membranes in biophysics, synthetic biology, and origins of life studies. Instead of the traditionally used Rayleigh-Gans-Debye approximation, we use an exact extended Lorenz-Mie solution for how core-shell particles scatter light to model vesicle turbidity. This approach enables accurate interpretations of simple turbidimetric measurements and is able to accurately model highly scattering vesicles, such as larger vesicles, those with multiple layers, and those with encapsulated material. We uncover several surprising features, including that vesicle lamellarity has a larger effect on sample turbidity than vesicle size and that the technique can be used to measure the membrane thickness of vesicles. We also examine potential misinterpretations of turbidimetry and discuss when measurements are limited by forward and multiple scattering and by the geometry of the instrument.

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“…All these advantages notwithstanding, turbidity spectrum analysis has attracted little attention to date, especially in the field of drug carrier research. Only a few practical characterization attempts were published to date (36)(37)(38)(39) and, at least for lipid vesicles, were quantitatively unsuccessful; this is possibly due to some underlying theoretical and/or experimental problems. We considered these problems and in this publication show how turbidity spectrum analysis can be successfully applied in the pharmaceutical field for the purpose of particulate drug carrier characterization.…”

Section: Introductionmentioning

confidence: 99%

“…APPENDIX D: THE REFRACTIVE INDEX OF LIPIDKhlebtsov and colleagues(37) proposed the following parametric description of dipalmitoylphosphatidylcholine (DPPC) refractive index as a function of light wavelength at 20°C, based on the data measured by Chong and Colbow with visible light above 400 nm (36): n L l ð Þ ¼ 1:4713 þ 1:31 nm l À1 þ 4309 nm 2 l À2 :…”

mentioning

confidence: 99%

Turbidity Spectroscopy for Characterization of Submicroscopic Drug Carriers, Such as Nanoparticles and Lipid Vesicles: Size Determination

Elsayed

Cevc²

2011

Pharm Res

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Studies of phosphatidylcholine vesicles by spectroturbidimetric and dynamic light scattering methods

Cited by 14 publications

References 19 publications

Development of Novel High-Resolution Size-Guided Turbidimetry-Enabled Particle Purification Liquid Chromatography (PPLC): Extracellular Vesicles and Membraneless Condensates in Focus

Development of Novel High-Resolution Size-Guided Turbidimetry-Enabled Particle Purification Liquid Chromatography (PPLC): Extracellular Vesicles and Membraneless Condensates in Focus

Core-Shell Modeling of Light Scattering by Vesicles: Effect of Size, Contents, and Lamellarity

Turbidity Spectroscopy for Characterization of Submicroscopic Drug Carriers, Such as Nanoparticles and Lipid Vesicles: Size Determination

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