Purification of HCC-specific extracellular vesicles on nanosubstrates for early HCC detection by digital scoring

The design and preparation of an EV click chip

An EV Click Chip (Fig. 1) is composed of two functional components: (i) Tz-grafted SiNWS: a patterned SiNWS32 covalently functionalized with disulfide bonds that link to terminal Tz motifs40, and (ii) an overlaid PDMS chaotic mixer41 (Supplementary Fig. 1), housed in a custom-designed microfluidic chip holder. The fabrication of Tz-grafted SiNWS began with introducing 10–15 µm densely packed Si nanowires (diameter = 100–200 nm) onto SiNWS, offering ~30 times more surface area (in contrast to a flat substrate) for facilitating click chemistry-mediated HCC EV capture. The incorporation of disulfide bonds and terminal Tz motifs onto SiNWS was carried out via a 3-step procedure40 (Supplementary Fig. 2). To confirm successful preparation of Tz-grafted SiNWS, X-ray photoelectron spectroscopy (XPS) was employed to monitor functional group transformation at each step9,40. The passive mixing behavior of the flow-through

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