Magnet Patterned Superparamagnetic Fe₃O₄/Au Core–Shell Nanoplasmonic Sensing Array for Label-Free High Throughput Cytokine Immunoassay

Rapid and accurate immune monitoring plays a decisive role in effectively treating immune-related diseases especially at point-of-care, where an immediate decision on treatment is needed upon precise determination of the patient immune status. Derived from the emerging clinical demands, there is an urgent need for a cytokine immunoassay that offers unprecedented sensor performance with high sensitivity, throughput, and multiplexing capability, as well as short turnaround time at low system complexity, manufacturability, and scalability. In this paper, a label-free, high throughput cytokine immunoassay based on a magnet patterned Fe₃O₄/Au core–shell nanoparticle (FACSNP) sensing array is developed. By exploiting the unique superparamagnetic and plasmonic properties of the core–shell nanomaterials, a facile microarray patterning technique is established that allows the fabrication of a uniform, self-assembled microarray on a large surface area with remarkable tunability and scalability. The sensing performance of the FACSNP microarray is validated by real-time detection of four cytokines in complex biological samples, showing high sensitivity (≈20 pg mL⁻¹), selectivity and throughput with excellent statistical accuracy. The developed immunoassay is successfully applied for rapid determination of the functional immunophenotype of leukemia tumor-associated macrophages, manifesting its potential clinical applications for real-time immune monitoring, early cancer detection, and therapeutic drug stratification toward personalized medicine.