SAXS for monoclonal antibodies batch crystallizations Patrick Larpent 1 Lorenzo Codan 2 , Jameson Bothe 3 , Dirk Stueber 4 , Paul Reichert 5 , Thierry Fischmann 5 , Yongchao Su 4 , Suzette Pabit 6 , Sudipta Gupta 6 and Aaron Cote 7 1 Department of Analytical Research and Development, MSD Werthenstein BioPharma GmbH, Industrie Nord 1, 6105 Schachen, Switzerland 2 Department of Process Research and Development, MSD Werthenstein BioPharma GmbH, Industrie Nord 1, 6105 Schachen, Switzerland 3 Department of Analytical Research and Development, Merck & Co., Inc., Kenilworth, New Jersey 07033, United States 4 Department of Analytical Research and Development, Merck & Co., Inc., Rahway, New Jersey 07065, United States 5 Department of Protein and Structural Chemistry, Merck & Co., Inc., Kenilworth, New Jersey 07033, United States 6 Department of Analytical Research and Development, Merck & Co., Inc., West Point, Pennsylvania 19486, United States 7 Department of Biologics Process Research and Development, Merck & Co., Inc., Kenilworth, New Jersey 07033, United States The large-scale crystallization of monoclonal antibodies (mAbs) represents an attractive approach to downstream drug substance (DS) processing as opposed to the more classic, costly solution manufacturing. 1,2 mAb crystallization holds great promises for simplifying and improving the purification of protein DS. 3-5 mAb crystalline suspensions additionally offer the opportunity to investigate highly concentrated drug product (DP) formulations 6,7 (e.g., for subcutaneous delivery) and are expected to be more stable at ambient conditions resulting in increased shelf-life stability. Highly concentrated formulations are currently very challenging for mAb solutions, which typically suffer from elevated viscosityand chemical instability together with the need of freezing temperature for storage. 8 While single crystals of mAbs have been widely investigated by single crystal x-ray diffraction, poly-crystals made from batch-crystallization and their bulk characterization remain more complex compared to that of traditional small molecule active pharmaceutical ingredients (APIs). The intrinsic properties of mAb molecules such as high molecular flexibility typically result in crystallites being shear sensitive and having a strong propensity to dehydration and crystallinity loss. Due to their large unit cells, bulk mAb crystals remains challenging to characterize with typical wide-angle scattering (WAXS) tools such as laboratory x-ray powder diffraction (XRPD). Small-angle x-ray scattering (SAXS) offers the ability to probe crystalline Bragg peaks at angles lower than conventional XRPD instruments enabling detection of x-ray diffractions from mAb crystals. In this presentation, we present how SAXS was used to support development of the crystallization process of an anti-PD1 mAb, currently administered as intravenous injection. We will describe how SAXS was used to (i) monitor batch crystallizations and confirm phase purity & crystallinity; (ii) assess crystallite stability in suspension and in the solid state, and; (iii) provide guidance on the best path forward for delivering crystalline pembrolizumab as both DS and DP. References 1. MAbs 2010 ; 2 :480-99 2. Biotechnol. Bioeng. 2013 ; 110 :2452-61 3. FEBS J. 2013 ; 280 :6456-6497 4. Cryst. Growth Des. 2012 ; 12 :6199-6207 5. Biotechnol. Adv. 2017 ; 35 :41-50 6. Expert Opin. Biol. Ther. 2004 ; 4 :301-17 7. PNAS 2003 ; 100 :6934-6939 8. J. Pharm. Sci. 2010 ; 99 :4812-29
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