Leurs Ulrike, Beck Hermann, Bonnington Lea, Lindner Ingo, Pol Ewa, Rand Kasper
Department of Pharmacy, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark.
Pharma Technical Development Basel, F. Hoffmann-La Roche Ltd., Grenzacherstrasse 124, 4070, Basel, Switzerland.
Chembiochem. 2017 Jun 1;18(11):1016-1021. doi: 10.1002/cbic.201600670. Epub 2017 May 3.
Protein-based pharmaceuticals represent the fastest growing group of drugs in development in the pharmaceutical industry. One of the major challenges in the discovery, development, and distribution of biopharmaceuticals is the assessment of changes in their higher-order structure due to chemical modification. Here, we investigated the interactions of three different biochemical probes (F s) generated to detect conformational changes in a therapeutic IgG1 antibody (mAbX) by local hydrogen-deuterium exchange mass spectrometry (HDX-MS). We show that two of the probes target the F part of the antibody, whereas the third probe binds to the hinge region. Through HDX-ETD, we could distinguish specific binding patterns of the F -binding probes on mAbX at the amino-acid level. Preliminary surface plasmon resonance (SPR) experiments showed that these domain-selective F probes are sensitive to conformational changes in distinct regions of a full-length therapeutic antibody upon oxidation.
基于蛋白质的药物是制药行业中发展最快的一类药物。生物制药的发现、开发和分销面临的主要挑战之一是评估由于化学修饰导致的其高级结构的变化。在这里，我们通过局部氢-氘交换质谱（HDX-MS）研究了三种不同的生化探针（Fs）与一种治疗性IgG1抗体（mAbX）构象变化检测之间的相互作用。我们发现其中两种探针靶向抗体的F部分，而第三种探针与铰链区结合。通过HDX-ETD，我们能够在氨基酸水平上区分F结合探针在mAbX上的特异性结合模式。初步的表面等离子体共振（SPR）实验表明，这些结构域选择性的F探针在氧化时对全长治疗性抗体不同区域的构象变化敏感。
