RESUMEN
Aim: The aim of this work was to investigate the correlation between anti drug antibody (ADA) induction and how different manufacturing processes of biopharmaceuticals affect the immunogenicity of the protein. This was done by testing four different batches of the same recombinant human protein in transgenic (Tg) mice. Methodology: Wild type (Wt) and human protein-transgenic (Tg) mice were challenged by repeated subcutaneous injections of four batches of a drug candidate protein, obtained by different purification methods. Differences between drug-specific IgG1, IgG2a, IgG2b, IgG3 and IgM antibody patterns produced in Tg vs. Wt mice were investigated and compared to the plasma cytokine profiles. A conventional ELISA was used as a reference method for ADA detection. Results: ADA responses detected in Tg mice were mainly of the IgG1 subclass and occurred only in significant response to the batch containing the highest level of proteins originating from the recombinant host cells. Wt mice, on the other hand, showed a combined IgG1/IgG2b response to all drug batches, except to the batch with the highest purity. The most pure batch failed to induce significant ADA in both Wt and Tg animals, suggesting host cell derived impurities to be a strong contributing factor to the antibody responses observed. Conclusion: Thus, an isolated IgG1 response in drug-tolerant Tg mice may serve as a potential biomarker of an immunological reaction to process-related impurities of the protein drug. In contrast, a combined IgG1/IgG2b-profile, as observed in immunoreactive Wt mice, more likely reflects a xeno-response.