Antibodies are essential components of the immune system with a wide range of molecular targets. They have been recognized as modalities for treating several diseases and more than 130 approved antibody-based therapeutics are available for clinical use. However, limitations remain associated with its efficacy, tissue permeability, and safety, especially in cancer treatment. Nanoparticles, particularly those responsive to external stimuli, have shown promise in improving the efficacy of antibody-based therapeutics and tissue-selective delivery. In this study, we developed a reliable and accurate method for quantifying the amount of antibody loaded onto lipid nanoparticles modified with Herceptin® (Trastuzumab), an antibody-based therapeutic used to treat HER2-positive cancers, using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by silver staining. This method proved to be a suitable alternative to commonly used protein quantification techniques, which are limited by lipid interference present in the samples. Furthermore, the amount of Herceptin modified on the liposomes, measured by this method, was confirmed by Herceptin's antibody-dependent cell-mediated cytotoxicity activity. Our results demonstrate the potential of this method as a critical tool for developing tissue-selective antibody delivery systems, leading to improved efficacy and reduced side effects of antibody-based therapeutics.

抗体是免疫系统的重要组成部分，具有广泛的分子靶标。它们已被认为是治疗多种疾病的方法，并且有超过 130 种经批准的基于抗体的疗法可供临床使用。然而，其功效、组织渗透性和安全性仍然存在局限性，特别是在癌症治疗中。纳米颗粒，特别是那些对外部刺激敏感的纳米颗粒，在提高基于抗体的治疗和组织选择性递送的功效方面显示出了希望。在这项研究中，我们开发了一种可靠且准确的方法，使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳来量化赫赛汀®（曲妥珠单抗）修饰的脂质纳米颗粒上的抗体量，赫赛汀是一种基于抗体的治疗剂，用于治疗 HER2 阳性癌症(SDS-PAGE)，然后进行银染色。该方法被证明是常用蛋白质定量技术的合适替代方法，该技术受到样品中存在的脂质干扰的限制。此外，通过该方法测量的脂质体上修饰的赫赛汀的量通过赫赛汀的抗体依赖性细胞介导的细胞毒性活性得到证实。我们的结果证明了该方法作为开发组织选择性抗体递送系统的关键工具的潜力，从而提高基于抗体的治疗的疗效并减少副作用。