ProteoDesign, Barcelona, Spain.
Methods Mol Biol. 2020;2133:183-200. doi: 10.1007/978-1-0716-0434-2_9.
Protein semisynthesis is a powerful tool for studying proteins and has contributed to a better understanding of protein structure and function and also driven innovations in protein science. Expressed protein ligation (EPL) is a widely used method to generate chemically modified proteins. However, EPL has some limitations, particularly relevant to modify challenging proteins such as antibodies. The method termed streamlined expressed protein ligation (SEPL) overcomes some of the problems of EPL, and other methods of protein semisynthesis, to generate challenging modified proteins such as antibody-drug conjugates (ADCs). ADCs targeting highly cytotoxic molecules to cancer cells, offer an attractive strategy to selectively eliminate tumor cells with improved therapeutic index than the antibodies or cytotoxic molecules themselves. Despite the potential of ADCs, the development of such complex molecules is challenging. We provide here protocols to prepare site-specifically modified ADCs by streamlined expressed protein ligation (SEPL), which does not require the incorporation of unnatural modifications into the antibody. Therefore, fully native antibodies, with only the desired cytotoxic molecules attached, can be generated.
蛋白质半合成是研究蛋白质的有力工具，有助于更好地理解蛋白质的结构和功能，并推动蛋白质科学的创新。表达蛋白连接（EPL）是一种广泛用于生成化学修饰蛋白的方法。然而，EPL 存在一些局限性，特别是在修饰具有挑战性的蛋白如抗体时更为明显。被称为简化表达蛋白连接（SEPL）的方法克服了 EPL 及其他蛋白半合成方法的一些问题，可用于生成具有挑战性的修饰蛋白，如抗体药物偶联物（ADC）。ADC 靶向高细胞毒性分子到癌细胞，提供了一种有吸引力的策略，与抗体或细胞毒性分子本身相比，选择性消除肿瘤细胞，提高治疗指数。尽管 ADC 具有潜力，但开发此类复杂分子具有挑战性。我们在此提供通过简化表达蛋白连接（SEPL）制备定点修饰 ADC 的方案，该方法不需要将非天然修饰物掺入抗体中。因此，可以生成仅附着所需细胞毒性分子的完全天然抗体。
