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使用其大小分布分析缓冲液中带电荷的巨大囊泡的纯化。

Analysis of purification of charged giant vesicles in a buffer using their size distribution.

机构信息

Department of Physics, Bangladesh University of Engineering and Technology (BUET), Dhaka, 1000, Bangladesh.

出版信息

Eur Phys J E Soft Matter. 2021 Apr 28;44(4):62. doi: 10.1140/epje/s10189-021-00071-4.

Abstract

We have analyzed the purification of charged giant unilamellar vesicles (GUVs) prepared in a buffer containing various concentrations of salt using their size distribution. The membranes of GUVs were synthesized by a mixture of dioleoylphosphocholine (DOPC) and dioleoylphosphatidylglycerol (DOPG) lipids. The DOPG mole fractions (X) in the membranes of GUVs were 0.10, 0.25, 0.40, 0.55, 0.70, 0.90 in a physiological buffer containing 162 mM salt. In addition, for a fixed value of X the concentrations of salt (C) in the buffer were 12, 62, 112, 162, 212, 312, 362 mM. The size distribution histograms of experimentally investigated unpurified and purified GUVs were fitted with the lognormal distribution and obtained the multiplication factor [Formula: see text] for mean ([Formula: see text]) and [Formula: see text] for standard deviation ([Formula: see text]) of the lognormal distribution. The key parameters [Formula: see text] and [Formula: see text] were responsible for changing the average size and size distribution of unpurified GUVs to purified ones. The theoretically fitting equation of experimentally obtained X- and C-dependent values of [Formula: see text] and [Formula: see text] provided the calibration equation for estimating the average size of purified GUVs theoretically for any values of X and C. The estimated size of purified GUVs increased with the increase in electrostatic effect (i.e., increase in vesicle surface charge density or decrease in salt concentration in buffer). The estimated size of purified GUVs varied with X and C, which supported the previous report qualitatively. These investigations might be helpful in the field of cell/chemical biology for understanding the process of purification of vesicles/cells investigated by any other techniques.

摘要

我们分析了在含有不同盐浓度的缓冲液中制备的带电巨单层囊泡(GUV)的纯化,方法是分析其大小分布。GUV 的膜由二油酰基磷酸胆碱(DOPC)和二油酰基磷脂酰甘油(DOPG)脂质的混合物合成。GUV 膜中的 DOPG 摩尔分数(X)在含有 162mM 盐的生理缓冲液中分别为 0.10、0.25、0.40、0.55、0.70、0.90。此外,对于固定的 X 值,缓冲液中的盐(C)浓度分别为 12、62、112、162、212、312、362mM。实验研究的未纯化和纯化 GUV 的大小分布直方图用对数正态分布拟合,并获得了对数正态分布的平均值[Formula: see text]的倍增因子和标准偏差[Formula: see text]的倍增因子。关键参数[Formula: see text]和[Formula: see text]负责将未纯化 GUV 的平均大小和大小分布改变为纯化 GUV。实验获得的 X 和 C 依赖性[Formula: see text]和[Formula: see text]的理论拟合方程为估计任何 X 和 C 值下纯化 GUV 的平均大小提供了校准方程。纯化 GUV 的估计大小随静电效应的增加而增加(即囊泡表面电荷密度的增加或缓冲液中盐浓度的降低)。纯化 GUV 的估计大小随 X 和 C 而变化,这从定性上支持了之前的报告。这些研究可能有助于细胞/化学生物学领域,有助于理解任何其他技术研究的囊泡/细胞的纯化过程。

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