title: Laser-induced breakdown detection of temperature-ramp generated aggregates of therapeutic monoclonal antibody.
abstract: The detection and characterization of protein aggregation is essential during development and quality control of therapeutic proteins, as aggregates are typically inactive and may trigger anti-drug-antibody formation in patients. Especially large multi-domain molecules, such as the important class of therapeutic monoclonal antibodies (mAbs), can form various aggregates that differ in size and morphology. Although particle analysis advanced over the recent years, new techniques and orthogonal methods are highly valued. To our knowledge, the physical principle of laser-induced breakdown detection (LIBD) was not yet applied to sense aggregates in therapeutic protein formulations. We established a LIBD setup to monitor the temperature-induced aggregation of a mAb. The obtained temperature of aggregation was in good agreement with the results from previously published temperature-ramped turbidity and dynamic light scattering measurements. This study demonstrates the promising applicability of LIBD to investigate aggregates from therapeutic proteins. The technique is also adaptive to online detection and size determination, and offers interesting opportunities for morphologic characterization of protein particles and impurities, which will be part of future studies.
authors.affiliation: Chair of Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-Universität München, Butenandtstrasse 5-13, D-81377 Munich, Germany.
authors.forename: Tim
authors.initials: T
authors.lastname: Menzen
authors.affiliation: Chair of Pharmaceutical Technology and Biopharmaceutics, Ludwig-Maximilians-Universität München, Butenandtstrasse 5-13, D-81377 Munich, Germany. Electronic address: wolfgang.friess@lrz.uni-muenchen.de.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Friess
authors.affiliation: Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, D-81377 Munich, Germany.
authors.forename: Reinhard
authors.initials: R
authors.lastname: Niessner
authors.affiliation: Chair of Analytical Chemistry, Technische Universität München, Marchioninistrasse 17, D-81377 Munich, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Haisch
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002152:Calorimetry, Differential Scanning; D002626:Chemistry, Pharmaceutical; D007834:Lasers; D066329:Protein Aggregates; D011487:Protein Conformation; D058767:Protein Unfolding; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D066329:Protein Aggregates
keywords: Laser-induced breakdown detection; Monoclonal antibody; Protein aggregation; Unfolding