An ELISA for detection of complement-bound circulating immune complexes in mice

Abstract

Measurements of complement-bound circulating immune complexes (cCICs) in pre-clinical studies may provide important information about the etiology of certain pathology findings suggestive of being immune complex mediated. This article describes the development and qualification of a universal methodology to measure cCIC in mice after dosing with species foreign proteins. The assay is a sandwich enzyme-linked immunosorbent assay – exclusively based on commercially available reagents – that could detect mouse IgG bound to complement C3 independent of the test-substance present in the plasma sample. Heat-aggregated serum was used as positive control. The assay was qualified by assessment of acceptance criteria, stability of positive control, precision, and specificity. Finally, the performance of the assay was tested using plasma from mice administered either of three different proteins, i.e bovine serum albumin (BSA), a fully human monoclonal antibody, and a humanized monoclonal antibody.

Keywords:

• Immunoassay
• immune complex
• complement 3 (C3)
• immunogenicity

Disclosure statement

The authors declare no conflicts of interest. The authors alone are responsible for the content of this article.

Additional information

Funding

This work was supported in part by Novo Nordisk A/S and the LifePharm Center for In Vivo Pharmacology at University of Copenhagen.