Immunogenicity of Structurally Perturbed Hen Egg Lysozyme Adsorbed to Silicone Oil Microdroplets in Wild-Type and Transgenic Mouse Models

Deep Convolutional Neural Network Analysis of Flow Imaging Microscopy Data to Classify Subvisible Particles in Protein Formulations

“…Figure generated with Matplotlib 48 and TikZ. 49 relevance of the classification [39][40][41] but also by the ease of generating data sets for classifier training with FIM images of only protein aggregates or silicone oil microdroplets. However, for many other relevant particle classification tasks, it is difficult to obtain data sets where each image is labeled as a member of a particle class that we would wish to differentiate from other particle classes.…”

Silicone Oil Particles in Prefilled Syringes With Human Monoclonal Antibody, Representative of Real-World Drug Products, Did Not Increase Immunogenicity in In Vivo and In Vitro Model Systems

“…For example, structurally modified lysozyme in the presence of SiOP was found to increase the immunogenic response in WT but not transgenic mice. 11 Further technical advances will allow more in-depth investigations to address additional questions that are outside the current scope of this report. For instance, additional IgG isotypes and other therapeutic modalities should be tested in the appropriately tolerized model systems analogous to the ones used here to determine the generality of our findings.…”

“…In contrast to our findings, recent reports regarding the safety of SiOP have shown some increased response in cell-or animal-based model immune systems challenged by mAb or recombinant protein added to stress-induced SiOP. [9][10][11]13 In most cases, emulsified silicone oil is spiked into the protein, or in others, stress-induced SiOP is mixed with mAb in buffers that are not representative of those used in commercial therapeutic protein formulations. It is unclear whether any denaturation of the protein caused by these conditions led to aggregates known to be mildly immunogenic, 18,24 consistent with mildly increased responses as compared to their negative or positive controls.…”

“…In one report that analyzed several interferon-b DPs, Avonex® that has the lowest rate of clinical immunogenicity (2%-13%) was found to contain high numbers of SiOP as compared to the other DPs, 16 indicating that SiOP does not pose an increased risk of immunogenicity in the clinic for these products. In other reports, an SiOP emulsion added to a monoclonal antibody (mAb) DP or lysozyme slightly enhanced cytokine secretion in peripheral blood mononuclear cells (PBMC) from a donor in vitro 13 or slightly increased antibodies in wild-type (WT) mice against the protein, 11 but did not increase ADA in transgenic mice.…”

“…Pharmaceutical industry along with regulatory agencies are concerned that SiOP has the potential to increase immunogenicity by functioning as an adjuvant or increasing protein aggregation. 4e7 Either of these mechanisms might generate anti-drug antibodies (ADA), [8][9][10][11] and thereby decrease the safety and efficacy of the drug. 6,12 Various recombinant proteins have been shown to adsorb, to different extents, on SiOP from emulsified silicone oil in the absence of surfactant, which is usually included in protein therapeutics to decrease undesirable interfacial and hydrophobic interactions.…”