TiO2-coated honeycomb-like super-macroporous silica for high-purity extraction of exosomes from human plasma

Abstract

Screening biomarkers from exosomes has emerged as a new strategy for non-invasive early diagnosis of diseases. Nevertheless, the present screening efficiency and accuracy of biomarkers are limited by the low extraction efficiency and purity of exosomes. To address this issue, a highly selective adsorbent, which integrates size-exclusion and chemisorption, was created by coating TiO₂ on honeycomb-like super-macroporous silica. Cell culture medium and human plasma were employed to investigate the enrichment performance, and the results indicate that the super-macropores ranging from 63.5 to 147.5 nm together with thin pore walls allow exosomes to enter and be adsorbed by TiO₂ in the pores, enhancing the available surface area for exosomes meanwhile physically excluding the large-sized cell debris and vesicles. Taking advantages of these properties, the prepared adsorbent achieves a higher extraction efficiency, recovery and purity of exosomes compared with the normal adsorbents and ultracentrifugation (UC) method. Combining this method with proteomic analysis, a total of 392 proteins were identified in exosomes from healthy human plasma, which is significantly higher than the number obtained by UC (200 proteins). For clinical samples, 59 upregulated and 124 downregulated proteins were identified in the plasma from colorectal cancer (CRC) patients, of which 44 upregulated proteins and 69 downregulated proteins are strongly associated with the progression of CRC. These findings suggest that this adsorbent possesses considerable potential in the extraction of exosomes for screening biomarkers and diagnosing tumor progress.