The successful development of targeted nanotherapeutics is contingent upon the conjugation of therapeutic nanoparticles to target-specific ligands, with particular emphasis being placed on antibody-based ligands. Thus, new methods that enable the covalent and precise installation of targeting antibodies to nanoparticle surfaces are greatly desired, especially those which do not rely on costly and time-consuming antibody engineering techniques. Herein we present a novel method for the highly controlled and oriented covalent conjugation of non-engineered antibody F(ab) fragments to PLGA–PEG nanoparticles using disulfide-selective pyridazinedione linkers and strain-promoted alkyne–azide click chemistry. Exemplification of this method with trastuzumab and cetuximab showed significant improvements in both conjugation efficiency and antigen binding capability, when compared to commonly employed strategies for antibody–nanoparticle construction. This new approach paves the way for the development of antibody-targeted nanomedicines with improved paratope availability, reproducibility and uniformity to enhance both biological activity and ease of manufacture.

中文翻译：

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通过非工程抗体片段的定向安装形成下一代抗体-纳米颗粒偶联物

靶向纳米治疗剂的成功开发取决于治疗性纳米颗粒与靶标特异性配体的缀合，尤其要重点放在基于抗体的配体上。因此，非常需要能够将靶向抗体共价且精确地安装到纳米颗粒表面的新方法，特别是那些不依赖于昂贵且费时的抗体工程技术的方法。本文中，我们提出了一种使用二硫键选择性哒嗪二酮连接子和应变促进炔-叠氮化物点击化学方法，将非工程化抗体F（ab）片段与PLGA-PEG纳米颗粒高度受控和定向共价偶联的新方法。与常用的抗体-纳米颗粒构建策略相比，用曲妥珠单抗和西妥昔单抗对该方法进行的示例显示出缀合效率和抗原结合能力均得到了显着改善。这种新方法为开发以抗体为靶点的纳米药物提供了途径，该药物具有更高的对位可用性，可重复性和均一性，从而增强了生物活性并易于制造。