Yamazoe Sayumi, Hogan Jason M, West Sean M, Deng Xiaodi A, Kotapati Srikanth, Shao Xiang, Holder Patrick, Lamba Vandana, Huber Mary, Qiang Cong, Gangwar Sanjeev, Rao Chetana, Dollinger Gavin, Rajpal Arvind, Strop Pavel
Discovery Biotherapeutics, Bristol-Myers Squibb, 700 Bay Road, Redwood City, California 94063, United States.
Protein Chemistry, Genentech Research and Early Development, 501 DNA Way, South San Francisco, California 94080, United States.
Bioconjug Chem. 2020 Apr 15;31(4):1199-1208. doi: 10.1021/acs.bioconjchem.0c00146. Epub 2020 Mar 27.
Antibody-drug conjugates (ADCs) are a therapeutic modality that traditionally enable the targeted delivery of highly potent cytotoxic agents to specific cells such as tumor cells. More recently, antibodies have been used to deliver molecules such as antibiotics, antigens, and adjuvants to bacteria or specific immune cell subsets. Site-directed mutagenesis of proteins permits more precise control over the site and stoichiometry of their conjugation, giving rise to homogeneous chemically defined ADCs. Identification of favorable sites for conjugation in antibodies is essential as reaction efficiency and product stability are influenced by the tertiary structure of immunoglobulin G (IgG). Current methods to evaluate potential conjugation sites are time-consuming and labor intensive, involving multistep processes for individually produced reactions. Here, we describe a highly efficient method for identification of conjugatable genetic variants by analyzing pooled ADC libraries using mass spectrometry. This approach provides a versatile platform to rapidly uncover new conjugation sites for site-specific ADCs.
抗体药物偶联物（ADCs）是一种治疗方式，传统上能够将强效细胞毒性药物靶向递送至特定细胞，如肿瘤细胞。最近，抗体已被用于将抗生素、抗原和佐剂等分子递送至细菌或特定免疫细胞亚群。蛋白质的定点诱变允许对其偶联的位点和化学计量进行更精确的控制，从而产生均一的化学定义明确的ADCs。由于免疫球蛋白G（IgG）的三级结构会影响反应效率和产物稳定性，因此确定抗体中有利的偶联位点至关重要。目前评估潜在偶联位点的方法既耗时又费力，涉及单独产生反应的多步过程。在此，我们描述了一种通过使用质谱分析汇集的ADC文库来鉴定可偶联遗传变体的高效方法。这种方法提供了一个通用平台，可快速发现位点特异性ADCs的新偶联位点。
