RT Journal Article
SR Electronic
T1 <em>In vitro</em> and <em>in vivo</em> immunogenicity assessment of protein aggregate characteristics
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.07.06.498969
DO 10.1101/2022.07.06.498969
A1 Camilla Thorlaksen
A1 Heidi S. Schultz
A1 Simon K. Gammelgaard
A1 Wim Jiskoot
A1 Nikos S. Hatzakis
A1 Flemming S. Nielsen
A1 Helene Solberg
A1 Vito Foderà
A1 Christina Bartholdy
A1 Minna Groenning
YR 2022
UL http://biorxiv.org/content/early/2022/07/07/2022.07.06.498969.abstract
AB The immunogenicity risk of therapeutic protein aggregates has been extensively investigated over the past decades. While it is established that not all aggregates are equally immunogenic, the specific aggregate characteristics which are most likely to induce an immune response, remain ambiguous. The aim of this study was to perform comprehensive in vitro and in vivo immunogenicity assessment of human insulin aggregates varying in size, structure and chemical modifications, while keeping other morphological characteristics constant. We found that flexible aggregates with highly altered secondary structure were most immunogenic in all setups, while compact aggregates with native-like structure were found to be immunogenic primarily in vivo. Moreover, sub-visible (1-100 µm) aggregates were found to be more immunogenic than sub-micron (0.1-1 µm) aggregates, while chemical modifications (deamidation, ethylation and covalent dimers) were not found to have any measurable impact on immunogenicity. The findings highlight the importance of utilizing aggregates varying in few characteristics for assessment of immunogenicity risk of specific morphological features and provides a universal workflow for reliable particle analysis in biotherapeutics.