Use of In Vitro Assays to Assess Immunogenicity Risk of Antibody-Based Biotherapeutics

Abstract

An In Vitro Comparative Immunogenicity Assessment (IVCIA) assay was evaluated as a tool for predicting the potential relative immunogenicity of biotherapeutic attributes. Peripheral blood mononuclear cells from up to 50 healthy naïve human donors were monitored up to 8 days for T-cell proliferation, the number of IL-2 or IFN-γ secreting cells, and the concentration of a panel of secreted cytokines. The response in the assay to 10 monoclonal antibodies was found to be in agreement with the clinical immunogenicity, suggesting that the assay might be applied to immunogenicity risk assessment of antibody biotherapeutic attributes. However, the response in the assay is a measure of T-cell functional activity and the alignment with clinical immunogenicity depends on several other factors. The assay was sensitive to sequence variants and could differentiate single point mutations of the same biotherapeutic. Nine mAbs that were highly aggregated by stirring induced a higher response in the assay than the original mAbs before stirring stress, in a manner that did not match the relative T-cell response of the original mAbs. In contrast, mAbs that were glycated by different sugars (galactose, glucose, and mannose) showed little to no increase in response in the assay above the response to the original mAbs before glycation treatment. The assay was also used successfully to assess similarity between multiple lots of the same mAb, both from the same manufacturer and from different manufacturers (biosimilars). A strategy for using the IVCIA assay for immunogenicity risk assessment during the entire lifespan development of biopharmaceuticals is proposed.

In silico and in vitro immunogenicity assessment of B‐domain‐modified recombinant factor VIII molecules

“…B‐domain junction peptides (Table ) were assessed for their ability to induce CD4 + T cell responses using the EpiScreen ™ time‐course T cell assay, performed by Antitope (now Abzena, Cambridge, UK). T cell proliferation and interleukin (IL)‐2 secretion were assessed using 3 H‐thymidine uptake and enzyme‐linked immunoSpot (ELISpot) assay, respectively.…”

Challenges and opportunities for the future of monoclonal antibody development: Improving safety assessment and reducing animal use

“…In part, the development of relevant in vitro assays with quantitative readouts in human primary cells and organs-on-chips will be instrumental in refining/applying these models for biologics. One successful example of an in vitro technique that has been applied to better characterize mAbs is the in vitro Comparative Immunogenicity Assessment (IVCIA) 30 . This was developed as a tool for predicting potential relative immunogenicity of biotherapeutic mAbs as a screening and prioritization tool, to differentiate mAbs and detect differential immunogenicity as a result of aggregation, which has been shown to potentially enhance cytokine secretion and T-cell proliferation response in healthy volunteers 31 …”

Cross‐Interaction Chromatography‐Based QSAR Model for Early‐Stage Screening to Facilitate Enhanced Developability of Monoclonal Antibody Therapeutics

“…With more mAbs being administered subcutaneously, higher concentration formulations of mAbs, when compared to mAbs administered intravenously, are required so as to administer higher amounts in smaller volumes (<1.5 mL of 100 mg doses) . The two main problems associated with high concentration mAb formulations are 1) increased aggregation propensities that could lead to immune‐related adverse effects and 2) decreased stability causing problems during administration and absorption in the subcutaneous space …”