Pharmaceutica Analytica Acta

Proteolysis inside a coated capillary: New development for the quality control of therapeutic antibodies

International Conference and Exhibition on Advances in HPLC & Chromatography Techniques

March 14-15, 2016 London, UK

Gaelle Coussot, Silvia Mas, Yoann Ladner, Killian Barkley and Catherine Perrin

Institut des Biomolécules Max Mousseron, France

Scientific Tracks Abstracts: Pharm Anal Acta

Abstract:

Monoclonal antibodies (mAb) represent the largest class of therapeutic molecules entering clinical studies. Due to their inherent structural complexity, the quality control (QC) of such type of molecules, necessary for their development and commercialization, represents a challenging analytical task. Common approaches in mAb QC are based on mAb reduction or proteolysis to produce a mixture of mAb fragments that are further analyzed by separation methods. These cleavage steps are usually performed off line (i.e. prior to the separation step in a distinct reactor). This is an important limitation in terms of time, reactant consumption and cross contamination by the possible formation of endogenous compounds that may further impair the quality assessment of the mAb drug. To overcome these limitations, we have first developed a fully integrated bio-analytical miniaturized methodology called D-PES (Diffusion�??mediated Proteolysis and Electrophoretic Separation) for the QC of polymer-drug conjugates. With the D-PES methodology, both cleavages and separation steps are performed in-line, silica capillary being used both as nanoreactor and separation support. The methodology is based on transverse diffusion of laminar flow profile (TDLFP) mixing of reactant nano-volumes (proteolytic buffer (PB), substrate (S), enzyme (E) or reducing agent (R)) inside the capillary. Principles and results of these rapid and low operating costs microanalyses will be presented for mAb drugs. Separation optimization and mAb cleavage conditions (choice of background electrolyte, PB, ionic strength, pH etc.) will be discussed to demonstrate the robustness of the D-PES methodology.

Biography :

Gaelle Coussot is Associate Professor at the Faculty of Pharmacy of Montpellier, France. She obtained her PhD in Analytical Chemistry in 2003. She then joined for 15 months the MD Anderson Center Cancer (Houston, Texas) for a Post-doctoral position in proteomic analyses. Currently, her researches focus on the development of bioanalytical methodologies using electrophoretic, chromatographic techniques and immunoassays to characterize and/or quantify proteins and others biopharmaceuticals. Research fields include quality control of biopharmaceuticals and study of antibodies resistance to particular environmental constraints. She has published 1 patent and 14 papers in international analytical and biochemical journals.

Email: gaelle.coussot@umontpellier.fr