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Page 50                                              Extracell Vesicles Circ Nucleic Acids 2020;1:20-56  I  http://dx.doi.org/10.20517/evcna.2020.10

               released from cells contained Cox suggesting recycling of protein or possibly recycling of entire EVs. Lastly,
               an assay was developed to measure functional EV uptake. Nanoluc protein was split in two and fused
               to mTurquoise2(N65) or mScarlet-I(66C). Expression of each fragment alone exhibited non-detectable
               levels of luminescence while expressing both together had a significantly increased signal. Delivery of
               either fragment within an EV to a cell expressing the corresponding fragment worked as confirmation and
               quantification of EV uptake (HEK293, U87, HeLa cells).
               Summary/Discussion: This study robustly demonstrates EV delivery of functional mRNA and protein to
               cells, while also establishing a simple assay to quantify and validate functional EV uptake.


               40. Smart Microprobes Imbued with Recognition Element as a Sensitive Bioanalysis Platform
               for Exosomes


                                                                          2
                                                1
               Authors: Chukwumaobim D. Nwokwu , Saif Mohammad Ishraq Bari , Gergana G. Nestorova 3
               E-mail: cdunwokwu@gmail.com
               Affiliations:
               1 Molecular Science and Nanotechnology, Louisiana Tech University, Ruston, LA, USA.
               2 Micro and Nanoscale Systems Engineering, Louisiana Tech University, Ruston, LA, USA.
               3 School of Biological Sciences, Louisiana Tech University, Ruston, LA, USA.
               Abstracts: Circulating exosomes have become useful biomarkers for precise and noninvasive diagnosis
               and disease monitoring. However, sample purity is a drawback for current liquid-phase methods for
               exosome isolation. We report a selective solid-phase technology for isolation of pure exosome populations.
               Microneedles (300 μm × 30 mm) were functionalized with exosome-specific anti-CD63 antibodies
               and their capture efficiency assessed via Fluorocet assay post-incubation in astrocyte-derived exosome
               suspension (EXO) enriched by a standard kit, as well as direct incubation in conditioned astrocyte medium
               (CAM), while blocking non-specific binding with 0.1% BSA in PBS. Our results indicated a 6-fold increase
               in exosomes captured by microprobes incubated overnight on ice in EXO (23 × 106 exosomes/probe) vis-
               à-vis 2 h incubation (3.0 × 106 exosomes/probe), and 2 folds more than the overnight probes at room
               temperature (12.9 × 106 exosomes/probe). The microprobe’s exosome loading capacity decreased when
               incubated in conditioned astrocyte medium, indicating that longer incubation at lower temperatures
               in enriched exosome suspension favors more efficient exosome capture. Our designed probe was also
               amenable to exosomal protein and miRNA extraction, in amounts sufficient for downstream analyses.
               Future works will focus on its integration into a lab-on-a-chip platform.


               41. Extracellular vesicles for drug delivery to the brain


                                         1
                                                                                            3
                                                                           2
                                                         2
               Authors: Bhagyashree S. Joshi , Marit A. De Beer , Ben N.G. Giepmans , Sameh A. Youssef , Alain de
                    3
                                       2
               Bruin , Harm H. Kampinga , Inge S. Zuhorn 1
               E-mail: i.zuhorn@umcg.nl
               Affiliations:
               1 University of Groningen, University Medical Center Groningen, Department of Biomedical Engineering,
               Groningen, The Nederlands.
               2 University of Groningen, University Medical Center Groningen, Department of Biomedical Sciences of Cells
               and Systems, Groningen, The Nederlands.
               3 University of Utrecht, Department of Biomolecular Health Sciences, Utrecht, The Nederlands.
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