Efficient methods to functionalize proteins are essential for the development of many diagnostic and therapeutic compounds, such as fluorescent probes for immunohistochemistry, zirconium-89 radiolabeled mAbs (⁸⁹Zr-mAbs) for positron emission tomography and antibody-drug conjugates (ADCs). This protocol describes a step-by-step procedure for the light-induced functionalization of proteins with compounds bearing the photochemically active aryl azide group. As an illustration of the potential utility of our approach, this protocol focuses on the synthesis of ⁸⁹Zr-mAbs using photoactivatable derivatives of the metal ion binding chelate desferrioxamine B (DFO). The light-induced synthesis of ⁸⁹Zr-mAbs is a unique, one-pot process involving simultaneous radiolabeling and protein conjugation. The photoradiochemical synthesis of purified ⁸⁹Zr-mAbs, starting from unmodified proteins, [⁸⁹Zr][Zr(C₂O₄)₄]^4– (⁸⁹Zr-oxalate), and a photoactivatable DFO derivative, can be performed in <90 min. The method can be easily adapted to prepare other radiolabeled proteins, ADCs or fluorescently tagged proteins by using drug molecules or fluorophores functionalized with photoactive moieties.

有效的蛋白质功能化方法对于许多诊断和治疗化合物的开发至关重要，例如用于免疫组织化学的荧光探针、用于正电子发射断层扫描的锆 89 放射性标记的 mAb ( ⁸⁹ Zr-mAb) 和抗体-药物偶联物 (ADC)。该协议描述了用带有光化学活性芳基叠氮化物基团的化合物对蛋白质进行光诱导功能化的分步程序。作为我们方法潜在效用的说明，该协议侧重于使用金属离子结合螯合物去铁胺 B (DFO) 的可光活化衍生物合成⁸⁹ Zr-mAb。⁸⁹的光诱导合成Zr-mAb 是一种独特的一锅法，涉及同时进行放射性标记和蛋白质缀合。从未修饰的蛋白质 [ ⁸⁹ Zr][Zr(C ₂ O ₄ ) ₄ ] ^4– ( ⁸⁹ Zr-草酸盐) 和可光活化的 DFO 衍生物开始，纯化的⁸⁹ Zr-mAb的光放射化学合成可以在 <90分钟 通过使用带有光活性部分功能化的药物分子或荧光团，该方法可以很容易地适用于制备其他放射性标记的蛋白质、ADC 或荧光标记的蛋白质。