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

Area of Science:

• Immunology
• Pre-clinical research

Background:

• Immune complex-mediated diseases are a significant concern in pre-clinical studies.
• Measuring complement-bound circulating immune complexes (cCICs) can offer insights into disease etiology.

Purpose of the Study:

• To develop and qualify a universal assay for measuring cCICs in mouse plasma.
• To enable the detection of mouse IgG bound to complement C3, irrespective of the test substance.

Main Methods:

• Development of a sandwich enzyme-linked immunosorbent assay (ELISA) using commercially available reagents.
• Qualification of the assay through evaluation of acceptance criteria, positive control stability, precision, and specificity.
• Testing the assay performance with plasma from mice treated with bovine serum albumin (BSA), a fully human monoclonal antibody, and a humanized monoclonal antibody.

Main Results:

• A universal methodology for measuring cCICs in mice was successfully developed and qualified.
• The assay demonstrated the ability to detect mouse IgG bound to complement C3.
• The assay's performance was validated in mice exposed to different protein antigens.

Conclusions:

• The developed universal assay provides a reliable tool for quantifying cCICs in pre-clinical mouse models.
• This assay can aid in the investigation of immune complex-mediated pathologies.
• The methodology is robust and applicable across various protein challenge models.