Asymmetric Flow Field-Flow Fractionation Investigation of Magnetopolyplexes

Haladjova, Emi; Rangelov, Stanislav; Geisler, Martin; Boye, Susanne; Lederer, Albena; Mountrichas, Grigoris; Pispas, Stergios

doi:10.1002/macp.201500177

Cited by 14 publications

(10 citation statements)

References 32 publications

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“…In contrast to HFF, AF4 (see the Supporting Information for details on the method) is based on much milder and gentler separation approach resulting in minimal destruction of supramolecular assemblies …”

Section: Resultsmentioning

confidence: 99%

Toward Functional Synthetic Cells: In‐Depth Study of Nanoparticle and Enzyme Diffusion through a Cross‐Linked Polymersome Membrane

et al. 2019

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Understanding the diffusion of nanoparticles through permeable membranes in cell mimics paves the way for the construction of more sophisticated synthetic protocells with control over the exchange of nanoparticles or biomacromolecules between different compartments. Nanoparticles postloading by swollen pH switchable polymersomes is investigated and nanoparticles locations at or within polymersome membrane and polymersome lumen are precisely determined. Validation of transmembrane diffusion properties is performed based on nanoparticles of different origin—gold, glycopolymer protein mimics, and the enzymes myoglobin and esterase—with dimensions between 5 and 15 nm. This process is compared with the in situ loading of nanoparticles during polymersome formation and analyzed by advanced multiple‐detector asymmetrical flow field‐flow fractionation (AF4). These experiments are supported by complementary i) release studies of protein mimics from polymersomes, ii) stability and cyclic pH switches test for in polymersome encapsulated myoglobin, and iii) cryogenic transmission electron microscopy studies on nanoparticles loaded polymersomes. Different locations (e.g., membrane and/or lumen) are identified for the uptake of each protein. The protein locations are extracted from the increasing scaling parameters and the decreasing apparent density of enzyme‐containing polymersomes as determined by AF4. Postloading demonstrates to be a valuable tool for the implementation of cell‐like functions in polymersomes.

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Section: Resultsmentioning

confidence: 99%

Toward Functional Synthetic Cells: In‐Depth Study of Nanoparticle and Enzyme Diffusion through a Cross‐Linked Polymersome Membrane

et al. 2019

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“…For homogeneous hard sphere the ratio is equal to 0.775, for linear random coil is between 1 and 2, and for elongated conformations the ratio is up to 2 [200]. The determination of the diffusion coefficient and thus the hydrodynamic radius is also possible from the AF4 elution time (see Section 5.1, Equation (3)); however, both approaches possess advantages and limitations that were treated in depth in the bioconjugation of synthetic macromolecules and proteins [177,[201][202][203][204][205].…”

Section: Determination Principle and Requirementsmentioning

confidence: 99%

Size Separation Techniques for the Characterisation of Cross-Linked Casein: A Review of Methods and Their Applications

et al. 2018

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Abstract:Casein is the major protein fraction in milk, and its cross-linking has been a topic of scientific interest for many years. Enzymatic cross-linking has huge potential to modify relevant techno-functional properties of casein, whereas non-enzymatic cross-linking occurs naturally during the storage and processing of milk and dairy products. Two size separation techniques were applied for characterisation of these reactions: gel electrophoresis and size exclusion chromatography. This review summarises their separation principles and discusses the outcome of studies on cross-linked casein from the last~20 years. Both methods, however, show limitations concerning separation range and are applied mainly under denaturing and reducing conditions. In contrast, field flow fractionation has a broad separation range and can be easily applied under native conditions. Although this method has become a powerful tool in polymer and nanoparticle analysis and was used in few studies on casein micelles, it has not yet been applied to investigate cross-linked casein. Finally, the principles and requirements for absolute molar mass determination are reviewed, which will be of increased interest in the future since suitable calibration substances for casein polymers are scarce.

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“…Currently, the most commonly used techniques for characterization of such systems are dynamic and electrophoretic light scattering as well as electron microscopy. Very few previous studies have attempted to obtain information about the detailed composition of polyplexes, their stoichiometry and distribution of the components utilizing powerful, informative but, in some aspects, exotic and not easily accessible techniques such as asymmetric flow field‐flow fractionation (AF4) and analytical ultracentrifugation (AUC) …”

Section: Introductionmentioning

confidence: 99%

Determination of Intimate Composition of Theranostic Polyplexes Based on (Co)Polymers of Poly(vinyl benzyl trimethylammonium chloride)

Haladjova

Mountrichas

Pispas

et al. 2017

Macro Chemistry & Physics

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Nonviral DNA CarriersNovel hybrid nanoparticulate systems that exhibit potential to combine therapeutic, diagnostic, and sensing modalities in a single nanoparticle are investigated. They are composed of a homopolymer of poly(vinyl benzyl trimethylammonium chloride) (or its copolymer with poly[oligo(ethylene glycol) methacrylate]), DNA, and gold nanoparticles (AuNPs). Using the approach of classic dynamic and static light scattering, parameters such as molar mass, particle size, geometry and density, and intimate composition are determined, trying to establish what the theranostic polyplexes really carry. According to the analysis, the polyplex particles are composed of up to 154 DNA molecules and 1612 (co)polymer molecules at amino-to-phosphate groups ratio (N/P) of 0.5 and 1 DNA and up to 252 (co)polymer molecules at higher N/P ratios. The particle morphology is consistent with "hairy surface on a compact sphere" with density that is lower than the density of the familiar copolymer micelles. The introduction of AuNPs does not influence the density and structure of the carriers, which could be related to the low number fraction of polyplex particles carrying AuNPs. Additional data from transmission electron microscopy, electrophoretic light scattering, and analytical ultracentrifugation validate the morphology, structure, and molar mass of the theranostic nanoassemblies and confirm the conclusions derived from light scattering.

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Asymmetric Flow Field-Flow Fractionation Investigation of Magnetopolyplexes

Cited by 14 publications

References 32 publications

Toward Functional Synthetic Cells: In‐Depth Study of Nanoparticle and Enzyme Diffusion through a Cross‐Linked Polymersome Membrane

Toward Functional Synthetic Cells: In‐Depth Study of Nanoparticle and Enzyme Diffusion through a Cross‐Linked Polymersome Membrane

Size Separation Techniques for the Characterisation of Cross-Linked Casein: A Review of Methods and Their Applications

Determination of Intimate Composition of Theranostic Polyplexes Based on (Co)Polymers of Poly(vinyl benzyl trimethylammonium chloride)

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