Coupling Nanostructured Microchips with Covalent Chemistry Enables Purification of Sarcoma‐Derived Extracellular Vesicles for Downstream Functional Studies

Abstract

Tumor-derived extracellular vesicles (EVs) play essential roles in intercellular communication during tumor growth and metastatic evolution. Currently, little is known about the possible roles of tumor-derived EVs in sarcoma because the lack of specific surface markers makes it technically challenging to purify sarcomaderived EVs. In this study, a specific purification system is developed for Ewing sarcoma (ES)-derived EVs by coupling covalent chemistry-mediated EV capture/ release within a nanostructure-embedded microchip. The purification platform-ES-EV Click Chip-takes advantage of specific anti-LINGO-1 recognition and sensitive click chemistry-mediated EV capture, followed by disulfide cleavagedriven EV release. Since the device is capable of specific and efficient purification of intact ES EVs with high purity, ES-EV Click Chip is ideal for conducting downstream functional studies of ES EVs. Absolute quantification of the molecular hallmark of ES (i.e., EWS rearrangements) using reverse transcription Droplet Digital PCR enables specific quantification of ES EVs. The purified ES EVs can be internalized by recipient cells and transfer their mRNA cargoes, exhibiting their biological intactness and potential role as biological shuttles in intercellular communication.

Importance of methylammonium iodide partial pressure and evaporation onset for the growth of co-evaporated methylammonium lead iodide absorbers

“…In recent years, perovskite solar cells (PSCs), have been subject to intense research due to the outstanding optoelectronic properties of the perovskite absorber 1 , 2 and the ease of fabrication through a variety of simple preparation methods 3 , 4 . Low processing temperatures 5 , 6 , high compositional versatility 7 – 11 and potential usage in cheap, high efficiency single- 12 , 13 as well as multi-junction (tandem) solar cells (SCs) 14 , 15 have further stimulated research interests.…”

“…Due to their low expense and simplicity in fabrication, up to now most groups have been using wet-chemical deposition approaches such as spin-coating in order to produce MAPbI layers for structural and optoelectronic analysis as well as photovoltaic applications 16 – 18 . Although up-scaling of wet-chemical deposition methods is being investigated 3 , 4 , their large scale preparation is still an open issue in terms of reproducibility, process yield and homogeneity 19 . On the other hand, the historic development of organic light emitting diodes (OLEDs) has shown that physical vapour deposition (PVD) is well suited for thin-film depositions at large scales and has great potential to succeed in the transition from laboratory production to industrial fabrication.…”

Advanced Nanotechnologies for Extracellular Vesicle‐Based Liquid Biopsy

“…[ 106 ] Similarly, Zhu and Tseng's team further developed a specific EV purification system by integrating covalent chemistry‐mediated EV capture on a Si nanowire‐based microfluidic chaotic mixer, enabling various downstream functional studies. [ 107 , 108 ] The colloidal self‐assembly (CSA) of nano SiO 2 is a promising approach to improve the specific surface area of the materials and to create different functionalized nanoscale interfaces. Zeng's team fabricated serpentine nanostructures on a bead pattern chip sealed by nano SiO 2 CSA and adopted a divarication microfluidic design (ExoProfile) integrated with various antibodies to enable multiplexed detection of biomarkers on EVs.…”

“…Recent studies have reported promising performance for EV isolation with ad hoc combinatorial EV capture instruments. [ 103 , 107 , 108 ] However, only a small fraction of these tools have been explored, suggesting that there is still sufficient room for developing highly efficient EV isolation procedures.…”

Nano Pom-Poms Prepared Exosomes for Highly Specific Cancer Biomarker Detection

“…Zhu etc. also reported the covalent chemistry mediated EV capture/release using click chip for EV puri cation and diagnosis of hepatocellular carcinoma (HCC) 28 and Ewing sarcoma 29 . The transcyclooctene (TCO) grafted antibody bound EVs with click chemistry motifs are presented after EV release which may introduce alteration of EV surface properties for in vivo applications.…”