Dickgiesser Stephan, Kellner Roland, Kolmar Harald, Rasche Nicolas
ADCs & Targeted NBE Therapeutics, Merck KGaA, Darmstadt, Germany.
Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany.
Methods Mol Biol. 2019;2033:1-14. doi: 10.1007/978-1-4939-9654-4_1.
Antibody-drug conjugates (ADCs) have been proven to be a successful therapeutic concept, allowing targeted delivery of highly potent active pharmaceutical ingredients (HPAPIs) selectively to tumor tissue. So far, HPAPIs have been mainly attached to the antibody via a chemical reaction of the payload with lysine or cysteine side chains of the antibody backbone. However, these conventional conjugation technologies result in formation of rather heterogeneous products with undesired properties. To overcome the limitations of heterogeneous ADC mixtures, several site-specific conjugation technologies have been developed over the last years. Originally pioneered by scientist from Genentech with their work on THIOMABs, several engineered cysteine mAb ADCs (ECM-ADCs) are now investigated in clinical trials. Here, we describe in detail how to engineer additional cysteines into antibodies and efficiently use them as highly site-specific conjugation sites for HPAPIs.
抗体药物偶联物（ADCs）已被证明是一种成功的治疗理念，能够将高效活性药物成分（HPAPIs）选择性地靶向递送至肿瘤组织。到目前为止，HPAPIs主要是通过有效载荷与抗体主链的赖氨酸或半胱氨酸侧链发生化学反应而连接到抗体上。然而，这些传统的偶联技术会导致形成具有不良性质的相当异质的产物。为了克服异质ADC混合物的局限性，在过去几年中已经开发了几种位点特异性偶联技术。最初由基因泰克的科学家在其关于硫醇化抗体（THIOMABs）的工作中开创，现在几种工程化半胱氨酸单克隆抗体ADC（ECM-ADCs）正在临床试验中进行研究。在此，我们详细描述如何将额外的半胱氨酸工程化到抗体中，并有效地将它们用作HPAPIs的高度位点特异性偶联位点。
