Isolation of Outer Membrane Vesicles Including Their Quantitative and Qualitative Analyses

Kohl, Paul; Zingl, Franz G.; Eichmann, Thomas O.; Schild, Stefan

doi:10.1007/978-1-4939-8685-9_11

Cited by 20 publications

(23 citation statements)

References 22 publications

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“…Effectively all HrpB in the supernatant was removed with ultracentrifugation, suggesting it is localized in OMVs ( Fig. 5C ) ( 36 ). Also, in the absence of HrpB coexpression, the levels of HrpC detected in the supernatant were reduced upon removal of insoluble material, while supernatant levels of HrpC detected after coexpression with HrpB were not affected by removal of insoluble material ( Fig.…”

Section: Resultsmentioning

confidence: 99%

A Widespread Bacterial Secretion System with Diverse Substrates

Grossman

Mauer

Forest

et al. 2021

mBio

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Section: Resultsmentioning

confidence: 99%

A Widespread Bacterial Secretion System with Diverse Substrates

Grossman

Mauer

Forest

et al. 2021

mBio

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“…In contrast, osmotic swelling of bacterial outer membrane vesicles provides a means to measure outer membrane permeability simply and directly in a bacterial membrane. Procedures for production and isolation of such OMVs have been well described [28,29], and they offer the advantage of a more physiological outer membrane (including LPS composition) than proteoliposomes [27]. Measuring osmotic swelling also permits the detection of a broader range of compounds than enzymatic detection approaches [8,30] and with less specialized apparatus than electrophysiological ones [24].…”

Section: Figurementioning

confidence: 99%

Antibiotic uptake across gram-negative outer membranes: better predictions towards better antibiotics

Ferreira

Kasson

2019

Preprint

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Crossing the gram-negative bacterial membrane poses a major barrier to antibiotic development, as many small molecules that can biochemically inhibit key bacterial processes are rendered microbiologically ineffective by their poor cellular uptake. The outer membrane is the major permeability barrier for many drug-like molecules, and the chemical properties that enable efficient uptake into mammalian cells fail to predict bacterial uptake. We have developed a computational method for accurate prospective prediction of outer-membrane uptake of drug-like molecules, which we combine with a new medium-throughput experimental assay. Parallel molecular dynamics simulations are used to successfully and quantitatively predict experimental permeabilities. For most polar molecules we test, outer membrane permeability also correlates well with whole-cell uptake. The ability to accurately predict and measure outer-membrane uptake of a wide variety of small molecules will enable simpler determination of which molecular scaffolds and which derivatives are most promising prior to extensive chemical synthesis. It will also assist in formulating a more systematic understanding of the chemical determinants of outer-membrane permeability.

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“…Vesicles are eventually collected by ultracentrifugation, ranging from 40,000 up to 175,000x g , depending on the bacterial species studied. Unfortunately, rotor type and centrifugation times are not specified in most papers, although these parameters are critical for yields of OMVs ( Kohl et al, 2018 ). Furthermore, ultracentrifugation alone may leave contaminants still present in the isolated OMV fraction.…”

Section: Induction and Isolation Of Omvs For Therapeutic Purposesmentioning

confidence: 99%

Outer Membrane Vesicle Induction and Isolation for Vaccine Development

2021

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Gram-negative bacteria release vesicular structures from their outer membrane, so called outer membrane vesicles (OMVs). OMVs have a variety of functions such as waste disposal, communication, and antigen or toxin delivery. These vesicles are the promising structures for vaccine development since OMVs carry many surface antigens that are identical to the bacterial surface. However, isolation is often difficult and results in low yields. Several methods to enhance OMV yield exist, but these do affect the resulting OMVs. In this review, our current knowledge about OMVs will be presented. Different methods to induce OMVs will be reviewed and their advantages and disadvantages will be discussed. The effects of the induction and isolation methods used in several immunological studies on OMVs will be compared. Finally, the challenges for OMV-based vaccine development will be examined and one example of a successful OMV-based vaccine will be presented.

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Isolation of Outer Membrane Vesicles Including Their Quantitative and Qualitative Analyses

Cited by 20 publications

References 22 publications

A Widespread Bacterial Secretion System with Diverse Substrates

A Widespread Bacterial Secretion System with Diverse Substrates

Antibiotic uptake across gram-negative outer membranes: better predictions towards better antibiotics

Outer Membrane Vesicle Induction and Isolation for Vaccine Development

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