Preparing giant unilamellar vesicles (GUVs) of complex lipid mixtures on demand: Mixing small unilamellar vesicles of compositionally heterogeneous mixtures

Bhatia, Tripta; Husen, Peter; Brewer, Jonathan R.; Bagatolli, Luís A.; Hansen, Per Lyngs; Ipsen, John Hjort; Mouritsen, Ole G.

doi:10.1016/j.bbamem.2015.09.020

Cited by 55 publications

(58 citation statements)

References 40 publications

(56 reference statements)

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“…This unmixing phenomena has been overcome in only limited ways, including electrofusion of single composition controlled-GUVs, 44 allowing for precise control at only the single assembly level and, more recently, by using carefully selected mixtures to form GUVs. 45 …”

Section: Resultsmentioning

confidence: 99%

Phase Composition Control in Microsphere-Supported Biomembrane Systems

Fried¹,

Gilchrist³

2017

Langmuir

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The popularization of studies in membrane protein lipid phase coexistence has prompted the development of new techniques to construct and study biomimetic systems with cholesterol-rich lipid microdomains. Here, microsphere supported biomembranes with integrated α-helical peptides, referred to as proteolipobeads (PLBs), were used to model peptide/protein partitioning within DOPC/DPPC/cholesterol phase-separated membranes. Due to the appearance of compositional heterogeneity and impurities in the formation of model PLB assemblies, fluorescence-activated cell sorting (FACS) was used to characterize and sort PLB populations on the basis of disordered phase (Ld) content. In addition, spectral imaging was used to assess the partitioning of FITC-labeled α-helical peptide between fluorescently labeled Ld phase and unlabeled ordered phase (Lo) phase lipid microdomains. The apparent peptide partition coefficient, Kp,app, was measured to be 0.89 ± 0.06, indicating a slight preference of the peptide for the Lo phase. A biomimetic motif of Lo phase concentration enhancement of biotinyl-peptide ligand display in proteolipobeads was also observed. Finally, peptide mobility was measured by FRAP separately in each lipid phase, yielding diffusivities of 0.036±0.005 and 0.014±0.003 µm2/s in the Ld and Lo phases, respectively.

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Section: Resultsmentioning

confidence: 99%

Phase Composition Control in Microsphere-Supported Biomembrane Systems

Fried¹,

Gilchrist³

2017

Langmuir

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“…Bhatia et al (2015) used mixture of different small unilamellar vesicles (SUVs) populations for obtain ternary GUV with uniform property. 9 …”

Section: Liposomes Preparationmentioning

confidence: 99%

Application of Various Types of Liposomes in Drug Delivery Systems

2017

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Liposomes, due to their various forms, require further exploration. These structures can deliver both hydrophilic and hydrophobic drugs for cancer, antibacterial, antifungal, immunomodulation, diagnostics, ophtalmica, vaccines, enzymes and genetic elements. Preparation of liposomes results in different properties for these systems. In addition, based on preparation methods, liposomes types can be unilamellar, multilamellar and giant unilamellar; however, there are many factors and difficulties that affect the development of liposome drug delivery structure. In the present review, we discuss some problems that impact drug delivery by liposomes. In addition, we discuss a new generation of liposomes, which is utilized for decreasing the limitation of the conventional liposomes.

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“…With diameters ranging from 1-200µm (111,118,143,(145)(146)(147), Giant Unilamellar Vesicles (GUVs) are similar in size to eukaryotic cells (148). The large size of GUVs renders techniques such as micromanipulation with optical tweezers or micropipettes, and direct observation by optical/fluorescence microscopy, possible (118,143,147).…”

Section: Giant Unilamellar Vesicles (Guvs)mentioning

confidence: 99%

“…However, they are more flexible and display an internal volume similar to that of cells (146) and are therefore considered promising cell mimetic system (148,150). The GUV membrane may comprise a single or a mixture of lipid classes (145) allowing their use in investigations into the function, shape, mechanical and chemical properties of complex biological membranes (145). They have been produced to contain IMPs (120,143,(151)(152)(153)(154)(155), fragments from a natural cell membranes (156,157) and soluble components within their lumen (158)(159)(160).…”

Section: Advantages Of Using Guvsmentioning

confidence: 99%

“…Fluorescent imaging has been used to observe lateral membrane organisation (175) and the micropipette aspiration technique has been employed with GUVs as cell membrane models (176,177). The bending stiffness of lipid bilayers and the free enrgies of interactions between membranes as well as membrane deformities (143), lipid membrane phase transitions (178) and membrane flickering (145,179) have also been investigated using GUVs.…”

Section: Advantages Of Using Guvsmentioning

confidence: 99%

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Methods of reconstitution to investigate membrane protein function

Skrzypek

Iqbal

Callaghan

2018

Methods

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Membrane proteins are notoriously difficult to investigate in isolation. The focus of this chapter is the key step following extraction and purification of membrane proteins; namely reconstitution. The process of reconstitution re-inserts proteins into a lipid bilayer that partly resembles their native environment. This native environment is vital to the stability of membrane proteins, ensuring that they undergo vital conformational transitions and maintain optimal interaction with their substrates. Reconstitution may take many forms and these have been classified into two broad categories. Symmetric systems enable unfettered access to both sides of a bilayer. Compartment containing systems contain a lumen and are ideally suited to measurement of transport processes. The investigator is encouraged to ascertain what aspects of protein function will be undertaken and to apply the most advantageous reconstitution system or systems. It is important to note that the process of reconstitution is not subject to defined protocols and requires empirical optimisation to specific targets.

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Preparing giant unilamellar vesicles (GUVs) of complex lipid mixtures on demand: Mixing small unilamellar vesicles of compositionally heterogeneous mixtures

Cited by 55 publications

References 40 publications

Phase Composition Control in Microsphere-Supported Biomembrane Systems

Phase Composition Control in Microsphere-Supported Biomembrane Systems

Application of Various Types of Liposomes in Drug Delivery Systems

Methods of reconstitution to investigate membrane protein function

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