Investigation of Protein Therapeutics in Frozen Conditions Using DNP MAS NMR: A Study on Pembrolizumab

• DOI: 10.1021/acs.molpharmaceut.4c00929
• Publication Date: 2024-11-18T14:26:31Z
• Authors: Daniel Banks; James G. Kempf; Yong Du; Paul Reichert; Chakravarthy Nara...; Rui Fang; Soonbum Kwon; Jing Ling; Ashley Lay-Forten...; Yongqian Zhang; Qing Zhe Ni; Aaron Cote; Yongchao Su

The success of modern biopharmaceutical products depends on enhancing the stability protein therapeutics. Freezing and thawing, which are common thermal stresses encountered throughout lifecycle drug substances, spanning production, formulation design, manufacturing, storage, shipping, can impact this stability. Understanding physicochemical molecular behaviors components in biological at temperatures relevant to manufacturing shipping is essential for assessing risks determining appropriate storage conditions. This study focuses high-concentration monoclonal antibody (mAb) pembrolizumab, substance Keytruda (Merck & Co., Inc., Rahway, NJ, United States), its excipients a frozen solution. By leveraging dynamic nuclear polarization (DNP), we achieve more than 100-fold signal enhancements solid-state NMR (ssNMR), enabling efficient low-temperature (LT) analysis pembrolizumab without isotopic enrichment. Through both ex situ ssNMR experiments conducted across temperature range 297 77 K, provide insights into crystalline under Importantly, utilizing LT magic-angle spinning (MAS) probes allows us dynamics from room down liquid nitrogen (<100 K). Our results demonstrate that valuable conformation dynamics, crystallinity, phase transformations during freezing matrix be readily obtained products. underscores potential LT-MAS DNP techniques analyzing therapeutics vaccines solutions.