NON-INVASIVE MONITORING CANCER USING INTEGRATED MICROFLUIDIC PROFILING OF CIRCULATING MICROVESICLES

摘要

A microfluidic exosome profiling platform integrating exosome isolation and targeted proteomic analysis is disclosed. This platform is capable of quantitative exosomal biomarker profiling directly from 30 μL plasma samples within approximately 100 minutes with markedly enhanced sensitivity and specificity. Identification of distinct subpopulation of patient-derived exosomes is demonstrated by probing surface proteins and multiparameter analyses of intravesicular biomarkers in the selected subpopulation. The expression of IGF-1R and its phosphorylation level in non-small cell lung cancer (NSCLC) patient plasma is assessed, as a non-invasive alternative to the conventional biopsy and immunohistochemistry. The microfluidic chip, which may be fabricated of a glass substrate and a layer of poly(dimethylsiloxane), can include a first capture chamber, a second capture chamber, a serpentine microchannel, a first microchannel, a second microchannel, a sample inlet, a buffer inlet, a bead inlet, at least a first connector channel, and a reagent inlet.

主权项

1. A microfluidic chip comprising:
a first capture chamber configured to enable immunomagnetic isolation; a second capture chamber configured to enable protein analysis; a serpentine microchannel connecting the first capture chamber and the second capture chamber; a first microchannel connecting the first capture chamber and the serpentine microchannel; a second microchannel connecting the serpentine microchannel and the second capture chamber; a sample inlet connected to the first capture chamber; a buffer inlet connected to the first capture chamber; a bead inlet; at least a first connector channel connected to the bead inlet and the first microchannel, wherein the first connector channel is connected to the first microchannel upstream of the serpentine microchannel and downstream of the first capture chamber; and a reagent inlet connected to the second microchannel upstream of the second capture chamber and downstream of the serpentine microchannel.