Low cost non-electromechanical technique for the purification of giant unilamellar vesicles

Karal, Mohammad Abu Sayem; Rahman, M. Mostafizur; Ahamed, Md. Kabir; Shibly, Sayed Ul Alam; Ahmed, Marzuk; Shakil, Md. Mostofa

doi:10.1007/s00249-019-01363-6

Cited by 23 publications

(25 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…simple but effective method has been used for preparing GUVs. 10,23,38,60,61 For the production of GUVs, 1,2-dioleoyl-snglycero-3-phospho-(1 0 -rac-glycerol) (sodium salt) (DOPG), 1,2dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dilauroyl-snglycero-3-phospho-(1 0 -rac-glycerol) (sodium salt) (DLPG), and 1,2-ditridecanoyl-sn-glycero-3-phosphocholine (DTPC) were purchased from Avanti Polar Lipids Inc. (Alabaster, AL). Bovine serum albumin (BSA), 1,4-piperazinediethanesulfonic acid (PIPES), ethylene glycol-bis(2-aminoethylether)-N,N,N 0 ,N 0 -tetraacetic acid (EGTA) and calcein were purchased from Sigma-Aldrich (Germany).…”

Section: Methodsmentioning

confidence: 99%

“…Cholesterol (chol) was purchased from WAKO pharmaceuticals (Japan). The GUVs were prepared in PIPES buffer (10 mM PIPES, pH 7.0, 150 mM NaCl and 1 mM EGTA) using the natural swelling method 59,60 Here, this method is described briey. A mixture of 1 mM DOPG and DOPC (about 200 mL) was placed in a glass vial and dried using a gentle ow of nitrogen gas to produce a thin and homogeneous lipid lm.…”

Section: Methodsmentioning

confidence: 99%

“…The size distribution of the unpuried and puried GUVs prepared using different conditions has been reported previously. 60,66 The size range for the unpuried GUVs was 3.0-60 mm, and the number of smaller vesicles with sizes of less than 10 mm was large. On the other hand, the size range for the puried GUVs was 7.0-60 mm, and the number of smaller vesicles with sizes of less than 10 mm was very small.…”

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Recent developments in the kinetics of ruptures of giant vesicles under constant tension

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Recent developments in the kinetics of ruptures of giant vesicles under constant tension

et al. 2021

Self Cite

View full text Add to dashboard Cite

show abstract

“…Those methods can purify the GUVs by eliminating the non-entrapped solute such as nanometer sized vesicles from the suspension. Recently, we have reported a gravity-based non-electromechanical membrane filtering technique [24], which shows similar performance like membrane filtering method. These methods/techniques provide a wide range size distribution of purified GUVs.…”

Section: Introductionmentioning

confidence: 93%

A new purification technique to obtain specific size distribution of giant lipid vesicles using dual filtration

et al. 2021

Self Cite

View full text Add to dashboard Cite

A new purification technique is developed for obtaining distribution of giant unilamellar vesicles (GUVs) within a specific range of sizes using dual filtration. The GUVs were prepared using well known natural swelling method. For filtration, different combinations of polycarbonate membranes were implemented in filter holders. In our experiment, the combinations of membranes were selected with corresponding pore sizes–(i) 12 and 10 μm, (ii) 12 and 8 μm, and (iii) 10 and 8 μm. By these filtration arrangements, obtained GUVs size distribution were in the ranges of 6−26 μm, 5–38 μm and 5–30 μm, respectively. In comparison, the size distribution range was much higher for single filtration technique, for example, 6−59 μm GUVs found for a membrane with 12 μm pores. Using this technique, the water-soluble fluorescent probe, calcein, can be removed from the suspension of GUVs successfully. The size distributions were analyzed with lognormal distribution. The skewness became smaller (narrow size distribution) when a dual filtration was used instead of single filtration. The mode of the size distribution obtained in dual filtration was also smaller to that of single filtration. By continuing this process of purification for a second time, the GUVs size distribution became even narrower. After using an extra filtration with dual filtration, two different size distributions of GUVs were obtained at a time. This experimental observation suggests that different size specific distributions of GUVs can be obtained easily, even if GUVs are prepared by different other methods.

show abstract

“…The elastic properties of membranes were measured by investigating the changes of volume in presence of Π using dynamic light scattering [ 5 , 6 ]. Since the static and dynamic change in size and shape of a ‘single GUV’ can be visualized using the optical microscope, vesicles with sizes 10 μm or more have been using in various researches [ 12 , 13 ]. When GUVs are transferred to a hypertonic solution, the shrinkage of GUVs occurs.…”

Section: Introductionmentioning

confidence: 99%

Effects of osmotic pressure on the irreversible electroporation in giant lipid vesicles

et al. 2021

Self Cite

View full text Add to dashboard Cite

Irreversible electroporation (IRE) is a nonthermal tumor/cell ablation technique in which a series of high-voltage short pulses are used. As a new approach, we aimed to investigate the rupture of giant unilamellar vesicles (GUVs) using the IRE technique under different osmotic pressures (Π), and estimated the membrane tension due to Π. Two categories of GUVs were used in this study. One was prepared with a mixture of dioleoylphosphatidylglycerol (DOPG), dioleoylphosphatidylcholine (DOPC) and cholesterol (chol) for obtaining more biological relevance while other with a mixture of DOPG and DOPC, with specific molar ratios. We determined the rate constant (kp) of rupture of DOPG/DOPC/chol (46/39/15)-GUVs and DOPG/DOPC (40/60)-GUVs induced by constant electric tension (σc) under different Π. The σc dependent kp values were fitted with a theoretical equation, and the corresponding membrane tension (σoseq) at swelling equilibrium under Π was estimated. The estimated membrane tension agreed well with the theoretical calculation within the experimental error. Interestingly, the values of σoseq were almost same for both types of synthesized GUVs under same osmotic pressure. We also examined the sucrose leakage, due to large osmotic pressure-induced pore formation, from the inside of DOPG/DOPC/chol(46/39/15)-GUVs. The estimated membrane tension due to large Π at which sucrose leaked out was very similar to the electric tension at which GUVs were ruptured without Π. We explained the σc and Π induced pore formation in the lipid membranes of GUVs.

show abstract

Low cost non-electromechanical technique for the purification of giant unilamellar vesicles

Cited by 23 publications

References 31 publications

Recent developments in the kinetics of ruptures of giant vesicles under constant tension

Recent developments in the kinetics of ruptures of giant vesicles under constant tension

A new purification technique to obtain specific size distribution of giant lipid vesicles using dual filtration

Effects of osmotic pressure on the irreversible electroporation in giant lipid vesicles

Contact Info

Product

Resources

About