Rationale: Therapeutic antibody conjugates allow for the specific delivery of cytotoxic agents or immune cells to tumors, thus enhancing the antitumor activity of these agents and minimizing adverse systemic effects. Most current antibody conjugates are prepared by nonspecific modification of antibody cysteine or lysine residues, inevitably resulting in the generation of heterogeneous conjugates with limited therapeutic efficacies. Traditional strategies to prepare homogeneous antibody conjugates require antibody engineering or chemical/enzymatic treatments, processes that often affect antibody folding and stability, as well as yield and cost. Developing a simple and cost-effective way to precisely couple functional payloads to native antibodies is of great importance. Methods: We describe a simple proximity-induced antibody conjugation method (pClick) that enables the synthesis of homogeneous antibody conjugates from native antibodies without requiring additional antibody engineering or post-synthesis treatments. A proximity-activated crosslinker is introduced into a chemically synthesized affinity peptide modified with a bioorthogonal handle. Upon binding to a specific antibody site, the affinity peptide covalently attaches to the antibody via spontaneous crosslinking, yielding an antibody molecule ready for bioorthogonal conjugation with payloads. Results: We have prepared well-defined antibody-drug conjugates and bispecific small molecule-antibody conjugates using pClick technology. The resulting conjugates exhibit excellent in vitro cytotoxic activity against cancer cells and, in the case of bispecific conjugates, superb antitumor activity in mouse xenograft models. Conclusions: Our pClick technology enables efficient, simple, and site-specific conjugation of various moieties to the existing native antibodies. This technology does not require antibody engineering or additional UV/chemical/enzymatic treatments, therefore providing a general, convenient strategy for developing novel antibody conjugates.

中文翻译：

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使用邻近诱导化学合成精密抗体偶联物。

基本原理：治疗性抗体偶联物允许将细胞毒剂或免疫细胞特异性递送至肿瘤，从而增强这些药物的抗肿瘤活性并最大限度地减少不良全身作用。目前大多数抗体偶联物是通过抗体半胱氨酸或赖氨酸残基的非特异性修饰制备的，不可避免地导致产生具有有限治疗效果的异质偶联物。制备均质抗体偶联物的传统策略需要抗体工程或化学/酶处理，这些过程通常会影响抗体折叠和稳定性，以及产量和成本。开发一种简单且具有成本效益的方法将功能有效载荷与天然抗体精确耦合非常重要。方法：我们描述了一种简单的邻近诱导抗体偶联方法 (pClick)，该方法能够从天然抗体合成同源抗体偶联物，而无需额外的抗体工程或合成后处理。将邻近激活的交联剂引入用生物正交手柄修饰的化学合成亲和肽中。在与特定抗体位点结合后，亲和肽通过自发交联共价结合到抗体上，从而产生一种抗体分子，可以与有效载荷进行生物正交偶联。结果：我们使用 pClick 技术制备了定义明确的抗体-药物偶联物和双特异性小分子-抗体偶联物。所得缀合物对癌细胞表现出优异的体外细胞毒活性，并且在双特异性缀合物的情况下，在小鼠异种移植模型中具有极好的抗肿瘤活性。结论：我们的 pClick 技术能够将各种部分高效、简单和位点特异性结合到现有的天然抗体上。该技术不需要抗体工程或额外的紫外线/化学/酶处理，因此为开发新型抗体偶联物提供了一种通用、方便的策略。