Antibodies have exquisite specificities for molecular recognition, which have led to their incorporation into array sensors that are crucial for research, diagnostic, and therapeutic applications. Many of these platforms rely heavily on surface-bound reactive groups to covalently tether antibodies to solid substrates; however, this strategy is hindered by a lack of orientation control over antibody immobilization. Here, we report a mild electrophilic phenylsulfonate (tosylate) ester-containing boronic acid affinity ligand for attaching antibodies to glass slides. A high level of antibody coupling located near the Fc region of the boronated antibody complex could be achieved by the proximal nucleophilic amino acid driven substitution reaction at the phenylsulfonate center. This enabled the full-length antibodies to be permanently tethered onto surfaces in an oriented manner. The advantages of this strategy were demonstrated through the individual and multiplex detection of protein and serum biomarkers. This strategy not only confers stability to the immobilized antibodies but also presents a different direction for the irreversible attachment of antibodies to solid supports in an orientation-controlled way.

中文翻译：

---

区域选择性 SN2 型反应用于将抗体定向和不可逆地固定在硼酸盐形成辅助的玻璃表面

抗体对分子识别具有精细的特异性，这导致它们被整合到对研究、诊断和治疗应用至关重要的阵列传感器中。其中许多平台严重依赖表面结合的反应基团将抗体共价连接到固体基质上。然而，该策略因缺乏对抗体固定的方向控制而受到阻碍。在这里，我们报告了一种温和的亲电子苯磺酸酯（甲苯磺酸酯）含硼酸亲和配体，用于将抗体连接到载玻片上。位于硼化抗体复合物 Fc 区附近的高水平抗体偶联可以通过苯磺酸盐中心的近端亲核氨基酸驱动取代反应来实现。这使得全长抗体能够以定向方式永久地拴在表面上。这种策略的优势通过蛋白质和血清生物标志物的单独和多重检测得到证明。这种策略不仅赋予了固定化抗体稳定性，而且为抗体以受控方向的方式不可逆地附着到固体支持物提供了不同的方向。