Application Notes

Getting the full picture: predicting the aggregation propensity of mAbs using chemical and thermal denaturation on a single, fully automated platform

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APPLICATION NOTE Prometheus is a trademark of NanoTemper Technologies GmbH, Munich, Germany. NanoTemper is a registered trademark of NanoTemper Technologies GmbH, Munich, Germany. ©2017 NanoTemper Technologies, Inc. South San Francisco, CA, USA. All Rights Reserved. NT-PR-011-2 nanotempertech.com References 1 Courtois F, Agrawal NJ, Lauer TM, Trout BL (2016) Rational design of therapeutic mAbs against aggregation through protein engineering and incorporation of glycosylation motifs applied to bevacizumab. mAbs 8: 99-112. 2 Garidel P, Karow A.R.; Blech, M. (2014) Orthogonal spectroscopic techniques for the early developability assessment of therapeutic protein candidates. Spectroscopy Europe 26: 9-13. 3 Geng SB, Cheung JK, Narasimhan C, Shameem M, Tessier PM (2014) Improving monoclonal antibody selection and engineering using measurements of colloidal protein interactions. Journal of pharmaceutical sciences 103: 3356-3363. 4 Homchaudhuri L, Kumar S, Swaminathan R (2006) Slow aggregation of lysozyme in alkaline pH monitored in real time employing the fluorescence anisotropy of covalently labelled dansyl probe. FEBS letters 580: 2097-2101. 5 Kumar S, Ravi VK, Swaminathan R (2009) Suppression of lysozyme aggregation at alkaline pH by tri-N-acetylchitotriose. Biochimica et biophysica acta 1794: 913-920. 6 Le Brun V, Friess W, Bassarab S, Garidel P (2010) Correlation of protein-protein interactions as assessed by affinity chromatography with colloidal protein stability: a case study with lysozyme. Pharmaceutical development and technology 15: 421-430. 7 Ravi VK, Swain T, Chandra N, Swaminathan R (2014) On the characterization of intermediates in the isodesmic aggregation pathway of hen lysozyme at alkaline pH. PloS one 9: e87256. 8 Rizzo JM, Shi S, Li Y, Semple A, Esposito JJ, Yu S, Richardson D, Antochshuk V, Shameem M (2015) Application of a high-throughput relative chemical stability assay to screen therapeutic protein formulations by assessment of conformational stability and correlation to aggregation propensity. Journal of pharmaceutical sciences 104: 1632-1640. 9 Roberts CJ (2014) Protein aggregation and its impact on product quality. Current opinion in biotechnology 30: 211-217. 10 Seeliger D, Schulz P, Litzenburger T, Spitz J, Hoerer S, Blech M, Enenkel B, Studts JM, Garidel P, Karow AR (2015) Boosting antibody developability through rational sequence optimization. mAbs 7: 505-515. 11 Temel DB, Landsman P, Brader ML (2016) Orthogonal Methods for Characterizing the Unfolding of Therapeutic Monoclonal Antibodies: Differential Scanning Calorimetry, Isothermal Chemical Denaturation, and Intrinsic Fluorescence with Concomitant Static Light Scattering. Methods in enzymology 567: 359-389.

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