Microfluidic device as a facile in vitro tool to generate and investigate lipid gradients

Neumann, Brittany M.; Kenney, Devin; Gericke, Arne

doi:10.1016/j.chemphyslip.2017.10.007

Cited by 2 publications

(1 citation statement)

References 50 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Biophysical experimental platforms that have the dynamic properties of the cellular plasma membrane and that can serve as intermediate fluidic devices between the existing solid supported bilayer platforms and complex cellular assays can provide valuable insight into viral infection. There are numerous reports of such model platforms that simplified various complex biochemical processes ( Rakowska et al, 2013 ; Jiang et al, 2014 ; Orosz et al, 2017 ; Neumann et al, 2018 ; Fonseka et al, 2019 ; Cavassin et al, 2020 ; Tang et al, 2021 ; Fonseka et al, 2022 ). In our previous study, we have investigated the relative affinity of the hemagglutinin ectodomain, the HA1 subunit, towards different GSLs ( Berselli et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

A cell free biomembrane platform for multimodal study of influenza virus hemagglutinin and for evaluation of entry-inhibitors against hemagglutinin

Roy

Byrne

Sarangi

et al. 2022

Front. Mol. Biosci.

View full text Add to dashboard Cite

Seasonal periodic pandemics and epidemics caused by Influenza A viruses (IAVs) are associated with high morbidity and mortality worldwide. They are frequent and unpredictable in severity so there is a need for biophysical platforms that can be used to provide both mechanistic insights into influenza virulence and its potential treatment by anti-IAV agents. Host membrane viral association through the glycoprotein hemagglutinin (HA) of IAVs is one of the primary steps in infection. HA is thus a potential target for drug discovery and development against influenza. Deconvolution of the multivalent interactions of HA at the interfaces of the host cell membrane can help unravel therapeutic targets. In this contribution, we reported the effect of a multivalent HA glycoprotein association on various glycosphingolipid receptors (GD1a, GM3, GM1) doped asymmetrically into an artificial host membrane spanned across an aqueous filled microcavity array. The extent of HA association and its impact on membrane resistance, capacitance, and diffusivity was measured using highly sensitive electrochemical impedance spectroscopy (EIS) and fluorescence lifetime correlation spectroscopy (FLCS). Furthermore, we investigated the inhibition of the influenza HA glycoprotein association with the host mimetic surface by natural and synthetic sialic acid-based inhibitors (sialic acid, Siaα2,3-GalOMe, FB127, 3-sialyl lactose) using electrochemical impedance spectroscopy and observe that while all inhibit, they do not prevent host binding. Overall, the work demonstrates the platform provides a label-free screening platform for the biophysical evaluation of new inhibitors in the development of potential therapeutics for IAV infection prevention and treatment.

show abstract

Section: Introductionmentioning

confidence: 99%

A cell free biomembrane platform for multimodal study of influenza virus hemagglutinin and for evaluation of entry-inhibitors against hemagglutinin

Roy

Byrne

Sarangi

et al. 2022

Front. Mol. Biosci.

View full text Add to dashboard Cite

show abstract

Engineering Phosphatidylserine Containing Asymmetric Giant Unilamellar Vesicles

McDonough,

Paratore,

Ketelhohn

et al. 2024

Membranes

View full text Add to dashboard Cite

The plasma membrane lipid distribution is asymmetric, with several anionic lipid species located in its inner leaflet. Among these, phosphatidylserine (PS) plays a crucial role in various important physiological functions. Over the last decade several methods have been developed that allow for the fabrication of large or giant unilamellar vesicles (GUVs) with an asymmetric lipid composition. Investigating the physicochemical properties of PS in such asymmetric lipid bilayers and studying its interactions with proteins necessitates the reliable fabrication of asymmetric GUVs (aGUVs) with a high degree of asymmetry that exhibit PS in the outer leaflet so that the interaction with peptides and proteins can be studied. Despite progress, achieving aGUVs with well-defined PS asymmetry remains challenging. Recently, a Ca2+-initiated hemifusion method has been introduced, utilizing the fusion of symmetric GUVs (sGUVs) with a supported lipid bilayer (SLB) for the fabrication of aGUVs. We extend this approach to create aGUVs with PS in the outer bilayer leaflet. Comparing the degree of asymmetry between aGUVs obtained via Ca2+ or Mg2+ initiated hemifusion of a phosphatidylcholine (PC) sGUVwith a PC/PS-supported lipid bilayer, we observe for both bivalent cations a significant number of aGUVs with near-complete asymmetry. The degree of asymmetry distribution is narrower for physiological salt conditions than at lower ionic strengths. While Ca2+ clusters PS in the SLB, macroscopic domain formation is absent in the presence of Mg2+. However, the clustering of PS upon the addition of Ca2+ is apparently too slow to have a negative effect on the quality of the obtained aGUVs. We introduce a data filtering method to select aGUVs that are best suited for further investigation.

show abstract

Microfluidic device as a facile in vitro tool to generate and investigate lipid gradients

Cited by 2 publications

References 50 publications

A cell free biomembrane platform for multimodal study of influenza virus hemagglutinin and for evaluation of entry-inhibitors against hemagglutinin

A cell free biomembrane platform for multimodal study of influenza virus hemagglutinin and for evaluation of entry-inhibitors against hemagglutinin

Engineering Phosphatidylserine Containing Asymmetric Giant Unilamellar Vesicles

Contact Info

Product

Resources

About