Controllable Assembly of Enzymes for Multiplexed Lab‐on‐a‐Chip Bioassays with a Tunable Detection Range

Abstract Multiplexed analysis of molecules with different concentrations requires assays with a tunable detection range. A strategy is outlined that uses click chemistry to assemble horseradish peroxidase in a controlled fashion to generate enzyme assemblies as probes for multiplexed bioassays. This controllable assembly of enzymes on detection antibodies allows for lab‐on‐a‐chip immunoassays with a tunable detection range from pg mL −1 to μg mL −1 . Simultaneous, multiplexed bioassays of clinically relevant inflammatory biomarkers in serum are demonstrated in one lab‐on‐a‐chip format, with a limit of detection of 0.47 pg mL −1 for interleukin‐6, 2.6 pg mL −1 for procalcitonin, and 40 ng mL −1 for C‐reactive protein. This controlled assembly technique provides a multiplexed platform for simultaneous and quantitative analyses of both low‐abundance and high‐abundance biomarkers with a broad detection range, which holds great promise as a point‐of‐care platform for biomedical diagnostics.