Integrated microreactor for enzymatic reaction automation: An easy step toward the quality control of monoclonal antibodies

Yoann Ladner (1) , Silvia Mas (1) , Gaelle Coussot (1) , Killian Bartley (1) , Jérôme Montels (1) , Jacques Morel (2) , Catherine Perrin (1)

1 IBMM - Institut des Biomolécules Max Mousseron [Pôle Chimie Balard]
2 Département de Rhumatologie[Montpellier]

The main purpose of the present work is to provide a fully integrated miniaturized electrophoretic methodology in order to facilitate the quality control of monoclonal antibodies (mAbs). This methodology called D-PES, which stands for Diffusion-mediated Proteolysis combined with an Electrophoretic Separation, permits to perform subsequently mAb tryptic digestion and electrophoresis separation of proteolysis products in an automated manner. Tryptic digestion conditions were optimized regarding the influence of enzyme concentration and incubation time in order to achieve similar enzymatic digestion efficiency to that obtained with the classical methodology (off-line). Then, the optimization of electrophoretic separation conditions concerning the nature of background electrolyte (BGE), ionic strength and pH was realized. Successful and repeatable electrophoretic profiles of three mAbs digests (Trastuzumab, Infliximab and Tocilizumab), comparable to the off-line digestion profiles, were obtained demonstrating the feasibility and robustness of the proposed methodology. In summary, the use of the proposed and optimized in-line approach opens a new, fast and easy way for the quality control of mAbs.

Mots clés: Quality Control; Antibodies; Monoclonal; Humanized; Automation; Capillary electrophoresis; Chemistry; Pharmaceutical; Electrophoresis; Capillary; Enzymatic microreactor; Enzyme Assays; Hydrogen-Ion Concentration; In-line digestion; Monoclonal antibody; Osmolar Concentration; Proteolysis; Proteolysis product separation; Quality control; Trastuzumab

Domaines: Sciences du Vivant [q-bio]

Yoann Ladner, Silvia Mas, Gaelle Coussot, Killian Bartley, Jérôme Montels, et al.. Integrated microreactor for enzymatic reaction automation: An easy step toward the quality control of monoclonal antibodies. Journal of Chromatography A, 2017, 1528, pp.83--90. ⟨10.1016/j.chroma.2017.10.066⟩. ⟨hal-01742108⟩