Pharmaceutical Candidate Optimization, Research and Development, Bristol-Myers Squibb Company, Princeton, NJ, USA.
Molecular Discovery Technologies, Research and Development, Bristol-Myers Squibb Company, Waltham, MA, USA.
J Am Soc Mass Spectrom. 2018 Jul;29(7):1524-1531. doi: 10.1007/s13361-018-1966-2. Epub 2018 May 7.
Higher-order structure (HOS) characterization of therapeutic protein-drug conjugates for comprehensive assessment of conjugation-induced protein conformational changes is an important consideration in the biopharmaceutical industry to ensure proper behavior of protein therapeutics. In this study, conformational dynamics of a small therapeutic protein, adnectin 1, together with its drug conjugate were characterized by hydrogen/deuterium exchange mass spectrometry (HDX-MS) with different spatial resolutions. Top-down HDX allows detailed assessment of the residue-level deuterium content in the payload conjugation region. HDX-MS dataset revealed the ability of peptide-based payload/linker to retain deuterium in HDX experiments. Combined results from intact, top-down, and bottom-up HDX indicated no significant conformational changes of adnectin 1 upon payload conjugation. Graphical Abstract ᅟ.
治疗性蛋白-药物偶联物的高级结构（HOS）表征对于全面评估偶联诱导的蛋白质构象变化是生物制药行业的重要考虑因素，以确保蛋白质治疗药物的正确行为。在这项研究中，通过氢/氘交换质谱（HDX-MS）以不同的空间分辨率对小治疗蛋白adnectin 1及其药物偶联物的构象动力学进行了表征。自上而下的 HDX 允许对有效载荷连接区域的残基水平氘含量进行详细评估。HDX-MS 数据集揭示了基于肽的有效载荷/连接子在 HDX 实验中保留氘的能力。完整的、自上而下的和自下而上的 HDX 的综合结果表明，在有效载荷偶联后，adnectin 1 没有发生明显的构象变化。
