Development of technologies for biomedical detection platform is critical to meet the global challenges of various disease diagnoses, especially for point-of-use applications. Because of its natural simplicity, effectiveness, and easy repeatability, random covalent-binding technique is widely adopted in antibody immobilization. However, its antigen-binding capacity is relatively low when compared to site-specific immobilization of antibody. Herein, we report that a detection platform modified with boronic acid (BA)-containing sulfobetaine-based polymer brush. Mainly because of the advantage of oriented immobilization of antibody endowed with BA-containing three-dimensional polymer brush architecture, the platform had a high antigen-binding capacity. Notably, nonspecific protein adsorption was also suppressed by the zwitterionic pendants, thus greatly enhanced signal-to-noise (S/N) values for antigen recognition. Furthermore, antibodies captured by BA pendants could be released in dissociation media. This new platform is promising for potential applications in immunoassays.