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Electrochemical Biosensor Targeting the Cancer Biomarker Human Ecto‐NOX Disulfide‐Thiol Exchanger 2 (ENOX2)

Human Ecto‐NOX Disulfide‐Thiol Exchanger 2 (ENOX2) is a cell‐surface metalocatalyst which is expressed solely on malignancies of all major human cancer types. ENOX2, along with its ubiquitous parent protein ENOX1, serve as terminal oxidases in plasma membrane electron transport chains, suggesting that such proteins are essential for cell growth. ENOX2 is frequently shed from malignancies into the sera and demonstrates site‐specific transcript variants, and thus presents an attractive target for both diagnostic blood testing and anti‐cancer therapeutic treatments. To better facilitate detection and targeting of ENOX2 in patient sera, we utilized SELEX to generate aptamers against recombinantly expressed ENOX2. These aptamers were characterized via gel mobility shift assays to determine the candidate aptamer with highest affinity for human ENOX2. The candidate with highest affinity was incorporated into an electrochemical DNA‐based biosensor, where changes in DNA confirmation upon target binding result in changed dynamics of an appended redox reporter molecule, ultimately generating a rapid and quantitative change in the current of the electrochemical response. This novel ENOX2‐targeting electrochemical biosensor allows rapid, dose‐responsive readout for the presence of ENOX2 in both buffer and blood serum.