We demonstrate a nano-sensing platform for the detection of cytokines from human cell lines and tissues sampled from patients with inflammatory bowel disease (IBD). Our nano-sensing platform facilitated binding between antigen and antibody, resulting in enhanced sensitivity compared to conventional approaches. This platform can be used to detect specific cytokine signals that reflect the condition of patients with IBD, where inflamed tissues result in increased sensing signal and vice versa. Significantly, we monitored the nuclear factor κB (NF-κB) abnormal activation, which control secretion of proinflammatory cytokines in human mucous membrane tissue. In addition, our nano-sensing platform can be used to measure abnormal activation of the NF-κB signaling pathway in macrophages and intestinal epithelial cells caused by, lipopolysaccharides and tumor necrosis factor α (TNF-α), respectively. Our method also enabled monitoring the anti-inflammatory effect of the IBD- drug treatment (lupeol), which inhibits the abnormal activation of NF-κB signal pathway. In contrast to conventional methods, we demonstrate that our covalent/non-covalent antibody biosensor can be utilized not only for establishing an in vivo model for IBD and clinical applications such as monitoring inflammation levels, but also for screening drug effects from human tissue and cell lines.