Impact of a Heat Shock Protein Impurity on the Immunogenicity of Biotherapeutic Monoclonal Antibodies

• Published in: Pharmaceutical research 2019-04, Vol.36 (4), p.51-14, Article 51
• Main Authors: Rane, Shraddha S., Dearman, Rebecca J., Kimber, Ian, Uddin, Shahid, Bishop, Stephen, Shah, Maryam, Podmore, Adrian, Pluen, Alain, Derrick, Jeremy P.
• Format: Article
• Language: English
• Subjects: Aggregates; Biochemistry; Biological products; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Biomedicine; Cell culture; Dendritic cells; Health aspects; Health risks; Heat shock proteins; Hsp70 protein; Immune response; Immunogenicity; Immunoglobulin G; Impurities; Lymphocytes; Lymphocytes T; Medical Law; Mice; Monoclonal antibodies; Pharmacology/Toxicology; Pharmacy; Protein binding; Proteins; Research Paper; Rodents; T cells; γ-Interferon
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1007/s11095-019-2586-7

Purpose Anti-drug antibodies can impair the efficacy of therapeutic proteins and, in some circumstances, induce adverse health effects. Immunogenicity can be promoted by aggregation; here we examined the ability of recombinant mouse heat shock protein 70 (rmHSP70) - a common host cell impurity - to modulate the immune responses to aggregates of two therapeutic mAbs in mice. Methods Heat and shaking stress methods were used to generate aggregates in the sub-micron size range from two human mAbs, and immunogenicity assessed by intraperitoneal exposure in BALB/c mice. Results rmHSP70 was shown to bind preferentially to aggregates of both mAbs, but not to the native, monomeric proteins. Aggregates supplemented with 0.1% rmHSP70 induced significantly enhanced IgG2a antibody responses compared with aggregates alone but the effect was not observed for monomeric mAbs. Dendritic cells pulsed with mAb aggregate showed enhanced IFNγ production on co-culture with T cells in the presence of rmHSP70. Conclusion The results indicate a Th1-skewing of the immune response by aggregates and show that murine rmHSP70 selectively modulates the immune response to mAb aggregates, but not monomer. These data suggest that heat shock protein impurities can selectively accumulate by binding to mAb aggregates and thus influence immunogenic responses to therapeutic proteins.