IMMUNOGENICITY OF THERAPEUTIC FUSION PROTEINS: CONTRIBUTORY FACTORS AND CLINICAL EXPERIENCE - Fusion Protein Technologies for Biopharmaceuticals: Applications and Challenges - page 83

FIGURE 5.4 Immunogenicity prediction strategy. A potential tiered approach to preclinical
immunogenicity testing is depicted here. Tier 1 would consist of the in silico screening of linear
sequences from multiple therapeutic candidates for T-cell epitopes and clusters. At the conclusion of
Tier 1, therapeutic candidates would be rated for immunogenic risk. Tier 2 would be an in vitro
evaluation of immunogenicity in a series of T-cell assays. This phase could include examination of
antigen processing and presentation, post-translational modifications, and different formulations.
Tier 3 would focus on immunogenicity testing of therapeutic candidates in vivo, in established
“humanized” animal models such as the Hu-SCID and HLA-transgenic mouse models. The
culmination of this immunogenicity prediction strategy would be advancement into the drug
development pipeline.

formulation such as interference from aggregates. Such
variables can enhance the immunogenicity of the drug
product in ways that cannot be captured by in silico
modeling, which only attends

sequence. Even in vitro tests cannot be fully indicative
of immunogenicity, and hence clinical evaluation of
immunogenicity is always required to confirm the devel-
opment predictions.

to the amino acid