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 \({\gamma }\) for mean (\({\mu }\)) and \({\eta }\) for standard deviation (\({\sigma }\)) of the lognormal distribution. The key parameters \(\gamma \) and \(\eta \) 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 \(\gamma \) and \(\eta \) 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.
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Data Availability Statement
This manuscript has no associated data or the data will not be deposited. [Authors’ comment: The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.]
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Acknowledgements
This work was supported partly by the Grants from Ministry of Science and Technology (Grant no: 39.00.0000.009.06.024.19-12), Ministry of Education (Grant no: 37.20.0000.004.033.020.2016), ICT Division (Grant no: 56.00.0000.028.33.105.18-05)(Ministry of Posts, Telecommunications and Information Technology) and CASR-BUET of Bangladesh to Mohammad Abu Sayem Karal.
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MA and MASK designed research. MA and MKA synthesized the samples, performed experiments and analytical calculations. MASK, MA and MSU wrote the paper.
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Ahmed, M., Karal, M.A.S., Ahamed, M.K. et al. Analysis of purification of charged giant vesicles in a buffer using their size distribution. Eur. Phys. J. E 44, 62 (2021). https://doi.org/10.1140/epje/s10189-021-00071-4
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DOI: https://doi.org/10.1140/epje/s10189-021-00071-4