Fig. 2: Optimization of EV Click Chips using artificial plasma samples. | Nature Communications

Fig. 2: Optimization of EV Click Chips using artificial plasma samples.

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

Fig. 2

a A quantitative method was developed for evaluating the performance of EV Click Chips using artificial plasma samples prepared by spiking HepG2 EVs into the plasma from a female healthy donor (HD). A RT-ddPCR assay was employed to quantify the copy numbers of the SRY and C1orf101 transcripts in the purified EV samples to calculate the recovery yield and recovery purity. *n is the ratio between C1orf101 and SRY transcripts in HepG2 EVs. b The recovery yields observed for EV Click Chips at different TCO-to-anti-EpCAM mole ratios. Data are presented as means ± SD of three independent assays. c–f The recovery yields obtained in the presence of individual and combined antibody capture agents, i.e., c anti-EpCAM, d anti-ASGPR1, e anti-CD147, and f combination of the three capture agents. Data are presented as means ± SD of three independent assays. g The recovery yields with different flow rates. Data are presented as means ± SD of three independent assays. h Dynamic ranges of EV recovery yields observed for EV Click Chips using artificial sample containing 0–9000 copies of SRY transcripts. Data are presented as means ± SD of three independent assays. i HepG2 EV recovery performance observed for (i) optimized EV Click Chips, devices without embedded silicon nanowires in SiNWS, and devices without herringbone features in the PDMS chaotic mixer, (ii) devices based on immunoaffinity EV capture (NanoVilli Chips) using the antibody cocktail concentration optimized for EV Click Chips, and (iii) ultracentrifugation (UC) approach. Data are presented as means ± SD of three independent assays. j General applicability of EV Click Chips for HCC EV recovery performance was validated using six artificial samples prepared by spiking three different HCC EVs (collected from HCC cell lines, i.e., HepG2, SNU387, and Hep3B) into two types of plasma samples (collected from either HD or liver cirrhotic patients). Data are presented as means ± SD of three independent assays.

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