A role for protein misfolding in immunogenicity of biopharmaceuticals

Publication Type:

Journal Article


Journal of Biological Chemistry, Volume 282, Number 4, pp. 2229-2236 (2007)



DOI Name (links to online publication)



cross-beta-structure; red-cell aplasia; human interferon alpha2b; immune tolerant mice; plasminogen activation; dendritic cells; wild-type; recombinant; therapeutics; aggregation


For largely unknown reasons, biopharmaceuticals evoke potentially harmful antibody formation. Such antibodies can inhibit drug efficacy and, when directed against endogenous proteins, cause life-threatening complications. Insight into the mechanisms by which biopharmaceuticals break tolerance and induce an immune response will contribute to finding solutions to prevent this adverse effect. Using a transgenic mouse model, we here demonstrate that protein misfolding, detected with the use of tissue-type plasminogen activator and thioflavin T, markers of amyloid-like properties, results in breaking of tolerance. In wild-type mice, misfolding enhances protein immunogenicity. Several commercially available biopharmaceutical products were found to contain misfolded proteins. In some cases, the level of misfolded protein was found to increase upon storage under conditions prescribed by the manufacturer. Our results indicate that misfolding of therapeutic proteins is an immunogenic signal and a risk factor for immunogenicity. These findings offer novel possibilities to detect immunogenic protein entities with tPA and reduce immunogenicity of biopharmaceuticals.