Lei Yutian, Shen Yuan, Chen Feng, He Rui, Zhang Zhang, Zhou Ying, Yu Jin-Chen, Crommen Jacques, Jiang Zhengjin, Wang Qiqin
Institute of Pharmaceutical Analysis, College of Pharmacy/State Key Laboratory of Bioactive Molecules and Druggability Assessment/International Cooperative Laboratory of Traditional Chinese Medicine Modernization and Innovative Drug Development of Ministry of Education (MOE) of China, Jinan University, Guangzhou 510632, China.
School of Biomedical Engineering, Sun Yat-sen University, Shenzhen 518107, China.
Acta Pharm Sin B. 2024 Nov;14(11):4962-4976. doi: 10.1016/j.apsb.2024.06.007. Epub 2024 Jun 19.
The dynamic tracking of antibody‒drug conjugates (ADCs) in serum is crucial. However, a versatile bioanalytical platform is lacking due to serious matrix interferences, the heterogeneity and complex biotransformation of ADCs, and the recognition deficiencies of traditional affinity technologies. To overcome this, a multiepitope recognition technology (MERT) was developed by simultaneously immobilizing CDR and non-CDR ligands onto MOF@AuNPs. MERT's excellent specificity, ultrahigh ligand density, and potential synergistic recognition ability enable it to target the different key regions of ADCs to overcome the deficiencies of traditional technologies. The binding capacity of MERT for antibodies is ten to hundred times higher than that of the mono-epitope or Fc-specific affinity technologies. Since MERT can efficiently capture target ADCs from serum, a novel bioanalytical platform based on MERT and RPLC‒QTOF-MS has been developed to monitor the dynamic changes of ADCs in serum, including the fast changes of drug-to-antibody ratio from 3.67 to 0.22, the loss of payloads (maytansinol), and the unexpected hydrolysis of the succinimide ring of the linker, which will contribute to clarify the fate of ADCs and provide a theoretical basis for future design. In summary, the MERT-based versatile platform will open a new avenue for in-depth studies of ADCs in biological fluids.
血清中抗体-药物偶联物（ADC）的动态跟踪至关重要。然而，由于严重的基质干扰、ADC的异质性和复杂的生物转化以及传统亲和技术的识别缺陷，缺乏一个通用的生物分析平台。为了克服这一问题，通过将互补决定区（CDR）和非CDR配体同时固定在金属有机框架负载金纳米颗粒（MOF@AuNPs）上，开发了一种多表位识别技术（MERT）。MERT出色的特异性、超高的配体密度和潜在的协同识别能力使其能够靶向ADC的不同关键区域，以克服传统技术的缺陷。MERT对抗体的结合能力比单表位或Fc特异性亲和技术高10到100倍。由于MERT能够从血清中高效捕获目标ADC，因此开发了一种基于MERT和反相高效液相色谱-四极杆飞行时间质谱（RPLC-QTOF-MS）的新型生物分析平台，以监测血清中ADC的动态变化，包括药物与抗体比率从3.67快速变化到0.22、有效载荷（美登素）的损失以及连接子琥珀酰亚胺环的意外水解，这将有助于阐明ADC的命运，并为未来的设计提供理论依据。总之，基于MERT的通用平台将为深入研究生物流体中的ADC开辟一条新途径。
