MAGEA4 Coated Extracellular Vesicles Are Stable and Can Be Assembled In Vitro

Reinsalu, Olavi; Samel, Anneli; Niemeister, Elen; Kurg, Reet

doi:10.3390/ijms22105208

Cited by 5 publications

(3 citation statements)

References 89 publications

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“…To produce enhanced green fluorescent protein (eGFP) loaded GUVs, MilliQ water-based IA solution was supplemented with 50 ng/μL of eGFP. The protocol for eGFP production and purification from the bacterial expression system has been described earlier 28 . The produced GUVs were imaged with a fluorescence microscope (EVOS M5000, Thermo Fisher Scientific Inc., USA).…”

Section: Methodsmentioning

confidence: 99%

Microfluidic production, stability and loading of synthetic giant unilamellar vesicles

Ernits,

Reinsalu,

Yandrapalli

et al. 2024

Sci Rep

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In advanced drug delivery, versatile liposomal formulations are commonly employed for safer and more accurate therapies. Here we report a method that allows a straightforward production of synthetic monodisperse (~ 100 μm) giant unilamellar vesicles (GUVs) using a microfluidic system. The stability analysis based on the microscopy imaging showed that at ambient conditions the produced GUVs had a half-life of 61 ± 2 h. However, it was observed that ~ 90% of the calcein dye that was loaded into GUVs was transported into a surrounding medium in 24 h, thus indicating that the GUVs may release these small dye molecules without distinguishable membrane disruption. We further demonstrated the feasibility of our method by loading GUVs with larger and very different cargo objects; small soluble fluorescent proteins and larger magnetic microparticles in a suspension. Compared to previously reported microfluidics-based production techniques, the obtained results indicate that our simplified method could be equally harnessed in creating GUVs with less cost, effort and time, which could further benefit studying closed membrane systems.

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Section: Methodsmentioning

confidence: 99%

Microfluidic production, stability and loading of synthetic giant unilamellar vesicles

Ernits,

Reinsalu,

Yandrapalli

et al. 2024

Sci Rep

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“…The MAGEA4 and 10 proteins are incorporated into nanosized biological vesicles, called extracellular vesicles (EVs), secreted into the conditioned cell culture media of transfected cells as well as COP5 cells expressing MAGEAs [25,26,30]. The relevance of this biological function of MAGEA proteins is not yet known, but we know that it is mediated by the C-terminal part of MAGEA4 containing the MHD region [25].…”

Section: Mutations Within the N-terminus Does Not Affect Magea10 Inco...mentioning

confidence: 99%

How the Intrinsically Disordered N-Terminus of Cancer/Testis Antigen MAGEA10 Is Responsible for Its Expression, Nuclear Localisation and Aberrant Migration

Samel,

Väärtnõu,

Verk

et al. 2023

Biomolecules

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Melanoma-associated antigen A (MAGEA) subfamily proteins are normally expressed in testis and/or placenta. However, aberrant expression is detected in the tumour cells of multiple types of human cancer. MAGEA expression is mainly observed in cancers that have acquired malignant phenotypes, invasiveness and metastasis, and the expression of MAGEA family proteins has been linked to poor prognosis in cancer patients. All MAGE proteins share the common MAGE homology domain (MHD) which encompasses up to 70% of the protein; however, the areas flanking the MHD region vary between family members and are poorly conserved. To investigate the molecular basis of MAGEA10 expression and anomalous mobility in gel, deletion and point-mutation, analyses of the MAGEA10 protein were performed. Our data show that the intrinsically disordered N-terminal domain and, specifically, the first seven amino acids containing a unique linear motif, PRAPKR, are responsible for its expression, aberrant migration in SDS-PAGE and nuclear localisation. The aberrant migration in gel and nuclear localisation are not related to each other. Hiding the N-terminus with an epitope tag strongly affected its mobility in gel and expression in cells. Our results suggest that the intrinsically disordered domains flanking the MHD determine the unique properties of individual MAGEA proteins.

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“…RNA content or miR levels were reduced by two cycles of freezing and thawing (Akers et al, 2016). Melanoma-associated antigen A (MAGEA)-coated extracellular vesicles can be freezenthawed twice without losing MAGEA4 in detectable amounts (Reinsalu et al, 2021). In recent, EDTA-free protease inhibitor cocktail was added to voided urine before storage at −80 °C and transcriptomics analysis for biomarker discovery in diabetic kidney disease (Barreiro et al, 2020).…”

Section: Storagementioning

confidence: 99%

Biological Features of Extracellular Vesicles and Challenges

Zeng

Qiu

Jiang

et al. 2022

Front. Cell Dev. Biol.

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Extracellular vesicles (EVs) are vesicles with a lipid bilayer membrane on the outside, which are widely found in various body fluids and contain biological macromolecules such as DNA, RNA, lipids and proteins on the inside. EVs were once thought to be vesicles for the removal of waste materials, but are now known to be involved in a variety of pathophysiological processes in many diseases. This study examines the advantage of EVs and the challenges associated with their application. A more rational use of the advantageous properties of EVs such as composition specificity, specific targeting, circulatory stability, active penetration of biological barriers, high efficient drug delivery vehicles and anticancer vaccines, oxidative phosphorylation activity and enzymatic activity, and the resolution of shortcomings such as isolation and purification methods, storage conditions and pharmacokinetics and biodistribution patterns during drug delivery will facilitate the clinical application of EVs.

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MAGEA4 Coated Extracellular Vesicles Are Stable and Can Be Assembled In Vitro

Cited by 5 publications

References 89 publications

Microfluidic production, stability and loading of synthetic giant unilamellar vesicles

Microfluidic production, stability and loading of synthetic giant unilamellar vesicles

How the Intrinsically Disordered N-Terminus of Cancer/Testis Antigen MAGEA10 Is Responsible for Its Expression, Nuclear Localisation and Aberrant Migration

Biological Features of Extracellular Vesicles and Challenges

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