Method for the determination of anti-drug antibodies against an effector function suppressed human or humanized drug antibody

Inventors

Umana, Pablo • Wessels, Uwe • Stubenrauch, Kay-Gunnar

Assignees

F Hoffmann La Roche AG • Roche Diagnostics GmbH • Hoffmann La Roche Inc

Abstract

Herein is reported an anti-drug antibody immunoassay for the determination of the presence of an anti-drug antibody against an effector function suppressed human or humanized drug antibody in a sample comprising the incubation of a sample comprising mammalian blood serum with full length human Fcgamma receptor I or an Fc-region binding fragment thereof so that a complex between the anti-drug antibody against the effector function suppressed human or humanized drug antibody present in the sample and the human Fcgamma receptor I or the Fc-region binding fragment thereof forms, whereby the full length human Fcgamma receptor I or the Fc-region binding fragment thereof is conjugated to a detectable label, and the determination of the formed complex by the detectable label.

Core Innovation

The invention provides an anti-drug antibody immunoassay for determining the presence of an anti-drug antibody against an effector function suppressed human or humanized drug antibody in a sample comprising mammalian blood serum. The immunoassay forms a complex between the anti-drug antibody present in the sample and full length human Fcgamma receptor I or an Fc-binding region of full-length human Fcgamma I, where the receptor or Fc-binding region is conjugated to a detectable label. The complex is isolated and determined by the detectable label.

In solid phase formats, the invention immobilizes the effector function suppressed human or humanized drug antibody, or its FAB, on a solid phase and incubates it with a sample comprising mammalian blood serum to form a drug antibody–anti-drug antibody complex or a FAB–anti-drug antibody complex. The solid phase is then incubated with full length human Fcgamma receptor I or an Fc-binding region conjugated to a detectable label, and the formation of a solid-phase-bound complex is determined by the presence of the detectable label.

Alternative workflows add excess drug antibody to transfer any anti-drug antibody present in the sample into a drug-antibody-anti-drug antibody complex, followed by solid-phase capture using an antigen specifically binding the effector function suppressed drug antibody, immobilized full length human Fcgamma receptor I or an Fc-binding region, or immobilized anti-drug antibody against the drug antibody. In each case, label-based detection with full length human Fcgamma receptor I or an Fc-binding region indicates the presence of an anti-drug antibody against an effector function suppressed human or humanized drug antibody.

Claims Coverage

The partial content includes six independent claims. The claims center on ADA detection for effector function suppressed human or humanized drug antibodies using human Fcgamma receptor I, full length or an Fc-binding region, conjugated to a detectable label, with multiple solid phase capture and ordered assay workflows.

Across the independent claims, the inventive features are directed to determining anti-drug antibody presence against effector function suppressed human or humanized drug antibodies by forming defined complexes and detecting them with human Fcgamma receptor I or an Fc-binding region conjugated to a detectable label, in solid phase and ordered assay workflows.

Stated Advantages

Documented Applications