The level of cortisol can reflect people's psychological stress, help diagnose adrenal gland diseases, and is also related to several mental diseases. In this study, we developed a cortisol monoclonal antibody-oriented approach to modify an immunosensor for wearable label-free and persistent sweat cortisol detection. On such an antibody-oriented immunosensor, the fragment crystallizable (Fc) region is partially inserted within the metal-organic framework (MOF), and antibody-binding regions of the cortisol monoclonal antibody (Cmab) were exposed on the MOF surface via selective growth and self-assembly. Such ordered and oriented embedding of antibodies in the MOF resulted in excellent antibody activity and improved stability and antigen-binding capacity. We also engineered the full integrated system for on-body sweat cortisol biosensing performance in several volunteers, and the results indicated that this wearable sensor is suitable for practical cortisol detection with a good linear detection range from 1 pg/mL to 1 μg/mL with a lower limit of detection of 0.26 pg/mL. Moreover, the wearable sensor demonstrated good persistence in detecting cortisol, with only 4.1% decay after 9 days of storage. The present work represents a simple oriented antibody assembling approach to improve the stability of antibodies, providing an important step toward long-term continuous sweat biomarker detection.