Background: Development of new recombinant biotechnology products has greatly expanded
in the field of modern pharmacy and medicine. Since biological recombinant molecules are
sensitive, simple or composed proteins, their function is heavily dependent on their structure. In addition
to their efficacy, biological medicinal products could show side effects such as immunogenicity.
Therefore, detection and characterization of protein structural variants is essential during development and
quality control of therapeutic proteins that might trigger immunogenic response in organism.
Methods: This article includes proposed detection and characterization of aggregated, as well as other
modified forms of monoclonal antibodies (mAb), by using selected chromatographic and spectrometric
methods. Additionally, selected mAb’s aggregates and modified structural variants of monoclonal antibodies
were subjected to the immature monocyte-derived dendritic cells’ (DC) examination experiment
for monitoring of activated DC cells in order to determine potential immunogenicity of mAb
structural variants. Furthermore, potential innate immunogenic response of peripheral blood mononuclear
cells (PBMC) cultures to mAb aggregates was also evaluated by measuring pro-inflammatory cytokine
response during early exposure of PBMCs to different mAb samples and by determining the effect
of mAb aggregates on PBMC proliferation during long-term cultures.
Result and Conclusion: All developed and proposed analytical methods and immunological in vitro
DC and PBMC assays, could be used as platform for complementary analytical characterization and
determination of potential for immunogenicity for all biopharmaceutical products which contain
monoclonal antibodies as active pharmaceutical ingredients.