title: Immuno-PCR assays for immunogenicity testing.
abstract: The administration of therapeutic proteins often induces immunogenic response and thus formation of anti-drug antibodies (ADA), which can neutralize the drug's therapeutic effect and may even cause serious health problems. We here report on the employment of the ultra-sensitive immuno-PCR (IPCR) method to facilitate immunogenicity testing using two established assay formats. In a "bridging assay", in which ADA forms a bridge to immobilize a signal-generating drug reporter probe, IPCR detection enabled an at least 1000-fold increase in sensitivity, as compared to the analogous ELISA, along with a high drug tolerance value. Moreover, we demonstrate that interfering effects of the biological matrix can be omitted by a simple dilution of analytical samples without loss in assay performance. In a cell-free "neutralizing assay", in which a labeled drug reporter probe competes for binding to either surface-bound receptors or neutralizing ADA, the IPCR assay also revealed high sensitivity. These results suggest that IPCR has the potential to become a standard methodology in immunogenicity testing.
authors: Chimera Biotec GmbH, D-44227 Dortmund, Germany. spengler@chimera-biotec.com
authors: Mark
authors: M
authors: Spengler
authors: Michael
authors: M
authors: Adler
authors: Andreas
authors: A
authors: Jonas
authors: Christof M
authors: CM
authors: Niemeyer
mesh_terms: D000906:Antibodies / Q000032:analysis*; D001710:Biotin; D004247:DNA / Q000032:analysis; D004341:Drug Evaluation / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D007108:Immune Tolerance; D004364:Pharmaceutical Preparations / Q000008:administration & dosage*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011506:Proteins / Q000008:administration & dosage / Q000276:immunology*
chemical_list: D000906:Antibodies; D004364:Pharmaceutical Preparations; D011506:Proteins; D001710:Biotin; D004247:DNA